Abstract:
The present invention aims at solving the problem of how to provide a starter culture, including a mixed starter culture, capable of fermenting e.g. grape juice without losing the desirable characteristics of less competitive non- Saccharomyces yeast  species early on in the fermentation process. In order to solve this problem there is provided in one aspect of the invention a composition comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces.  In another aspect of the invention there is provided a composition comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one second yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces.  In a further aspect of the invention there is provided a composition comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one further yeast organism selected from the genus of  Saccharomyces.  In yet another aspect of the invention there is provided a composition comprising at least one first yeast organism selected from the family of Saccaromycetaceae excluding the genus of  Saccharomyces  and at least one second yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one further yeast organism selected from the genus of  Saccharomyces.

Description:
TECHNICAL FIELD  
       [0001]     The present invention relates to yeast compositions, in particular yeast starter cultures capable of fermenting carbohydrate sources. The compositions can comprise one or more yeast organisms belonging to the family of Saccharomycetaceae, preferably selected from the geni of  Torulaspora  and/or  Kluyveromyces  and excluding the genus of  Saccharomyces,  optionally in combination with one or more yeast organisms belonging to the genus of  Saccharomyces.  The compositions are preferably freeze dried or spray/fluid bed dried. The invention also pertains to uses of the compositions for fermenting a carbohydrate source and producing an edible or drinkable product.  
       BACKGROUND OF THE INVENTION  
       [0002]     Grape juice is a very specialised ecosystem wherein only a few yeast species survive and proliferate. Examples of such few yeast species include the wild yeast families  Brettanomyces, Debaromyces, Hanseniaspora, Kloeckera, Torulaspora Zygo - Saccharomyces  and  Kluyveromyces  (Bauer &amp; Pretorius: S. Afr. J. Enol. Vitic. Vol. 21, special issue 2000, pp. 27-51). Survival and proliferation may be either desirable or undesirable depending on the yeast species in question. As grape juice is an oxygen limited and acidic environment with a high concentration of carbohydrates, these conditions naturally favour some yeasts species, which will tend to outgrow yeast species having different requirements for optimal growth. In order to obtain a wine quality with a refined aroma and flavour the use of starter cultures that contain a mixture of selected yeast species, may be considered for a person skilled in the art.  
         [0003]     One option for a person skilled in the art to obtain a desired ecosystem, which meets the desire of enhancing the growth of those yeast species that have desirable characteristics in terms of aroma and flavour, would be to compile these desirable yeast species that are able to outgrow the less desirable yeast species and let them make up the desired ecosystem. This ecosystem would be further improvable by addition of sulfur dioxide in antimicrobially effective amounts in order to inhibit or suppress the indigenous population of e.g. spoilage microorganisms.  
         [0004]      Saccharomyces cerevisiae  is capable of carrying out a final ethanol production due to its extreme adaptability to low oxygen concentration and high ethanol concentration, while other yeast species which may occur in combination with  S. cerevisiae,  only grow in a few days before they are outgrown by  S. cerevisiae.  Consequently, such other yeast species can only be found in young wine and they do not contribute to the fermentation process for more than a few days. As reported by Hansen et al. (J. Applied Microbiology; 2001, 91, 541-547), non- Saccharomyces  yeast species such as  Torulaspora delbrueckii  and  Kluyveromyces thermotolerans  are examples of yeast species, whose ability to grow is very sensitive to low oxygen contents of the growth medium. Such species are therefore not capable of participating for a sufficiently long time in a fermentation wherein  S. cerevisiae  species are also taking part. This implies a limitation of choice of culture to make up the composition of wine starter cultures and this limitation has for long been a serious problem to wine makers wishing to employ mixed yeast starter cultures for wine production. The background for wishing to use mixed yeast starter cultures may be a desire to produce a wine having a more refined aroma or flavour.  
         [0005]     It is known by wine makers that by spontaneous alcohol fermentation a more complex wine can often be produced, i.e. a wine with a desired broader spectrum of aroma and flavours than what is routinely achieved by using commercial starter cultures consisting exclusively of  S. cerevisiae.  However, such a spontaneous fermentation is very difficult to control and the internal competition between indigenous species present in the grape juice unfortunately often results in the dominance of undesired off-flavour producing species.  
         [0006]     Thus, while spontaneous fermentations in some cases may be more desirable than fermentations based on  S. cerevisiae  starter cultures, spontaneous fermentations are difficult to control and therefore wine with different and unknown characteristics from batch to batch result. In order to obtain a better control of the fermentation process many wine makers today rely on industrially produced  S. cerevisiae  starter cultures, which unfortunately are devoid of those yeast species which are capable of generating other desirable aroma and flavours. Even when such strains are present in combination with  S. cerevisiae  they are quickly outgrown due to a lesser survival tolerance-towards e.g. low oxygen levels and high ethanol concentrations.  
       SUMMARY OF THE INVENTION  
       [0007]     The present invention comprises the aim of solving the problem of how to provide a starter culture, including a mixed starter culture, capable of fermenting e.g. grape juice without losing the desirable characteristics of less competitive non- Saccharomyces  yeast species that grow vigorously early on in the fermentation process.  
         [0008]     In order to solve this problem, the artisans of the present invention have surprisingly provided in one aspect of the invention a starter culture as a composition of cultures comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces.    
         [0009]     In a second aspect of the invention there is provided a composition comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one second yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces.    
         [0010]     In a third aspect of the invention there is provided a composition comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one further yeast organism selected from the genus of  Saccharomyces.    
         [0011]     In a fourth aspect of the invention there is provided a composition comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one second yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one further yeast organism selected from the genus of  Saccharomyces.    
         [0012]     The invention also pertains to methods for preparing the above compositions as well as to methods for their use, including the use of the compositions as starter cultures for the fermentation of a carbohydrate source during e.g. wine making.  
         [0013]     Accordingly, in yet further aspects of the invention there is provided:  
         [0014]     A method for fermenting at least one fermentable carbohydrate source in an aqueous composition, said method comprising the step of contacting the aqueous composition with the composition according to the invention under conditions allowing said fermentation to occur.  
         [0015]     A method for producing an edible or drinkable product, said method comprising the step of adding the composition according to the invention to a precursor composition of said product, fermenting said precursor composition and producing said edible or drinkable product.  
         [0016]     A method for flavouring an edible or drinkable product, said method comprising the step of adding the composition according to the invention to a precursor composition of said product, fermenting said precursor composition and flavouring said edible or drinkable product.  
         [0017]     A method for preparing a dried composition according to the invention, said method comprising the steps of i) providing at least one first yeast organism, and ii) drying said at least one first yeast organism to produce a dried composition thereof.  
         [0018]     A method for preparing a dried composition according to the invention, said method comprising the steps of i) providing at least one first yeast organism selected from the genus of  Torulaspora  or the genus of  Kluyveromyces,  and ii) drying said composition.  
         [0019]     A method for preparing a dried composition according to the invention, said method comprising the steps of i) providing a composition comprising at least one first yeast organism selected from the genus of  Torulaspora  and at least one second yeast species selected from the genus of  Kluyveromyces,  and ii) drying said composition.  
         [0020]     A method for preparing a dried composition according to the invention, said method comprising the steps of i) providing a composition comprising at least one first yeast organism selected from the genus of  Torulaspora  or the genus of  Kluyveromyces,  and at least one further yeast organism selected from the genus of  Saccharomyces,  and ii) drying said composition.  
         [0021]     A method for preparing a dried composition according to the invention, said method comprising the steps of i) providing at least one first yeast organism selected from the genus of  Torulaspora  and at least one second yeast organism selected from the genus of  Kluyveromyces  and at least one further yeast organism selected from the genus of  Saccharomyces,  and ii) drying said composition.  
         [0022]     Use of a composition according to the invention for the manufacture of a yeast starter culture for fermenting grape juice.  
         [0023]     Use of a composition according to the invention for producing an edible or drinkable product.  
         [0024]     Use of a composition according to the invention for flavouring an edible or drinkable product.  
         [0025]     Use of at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  for the preparation of a yeast starter culture for fermenting grape juice and flavouring an edible or drinkable product resuiting from said fermentation.  
         [0026]     Use of a composition comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one second yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  for the preparation of a yeast starter culture for fermenting grape juice and flavouring an edible or drinkable product resulting from said fermentation.  
         [0027]     Use of a composition comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one further yeast organism selected from the genus of  Saccharomyces  for the preparation of a yeast starter culture for fermenting grape juice and flavouring an edible or drinkable product resulting from said fermentation.  
         [0028]     Use of a composition comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one second yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one further yeast organism selected from the genus of  Saccharomyces  for the preparation of a yeast starter culture for fermenting grape juice and flavouring an edible or drinkable product resulting from said fermentation.  
         [0000]     Definitions  
         [0029]     Starter culture: If not specifically defined otherwise, starter cultures are a yeast culture, which has been prepared for initiation of a fermentation and/or a conversion of edible or drinkable products.  
         [0030]     Yeast organism: Any genus belonging to the family of Saccharomycetaceae. Examples include, but are not limited to:  Saccharomyces, Kluyveromyces, Torulaspora, Arxiozyma, Citeromyces, Debaryomyces, Dekkera, Holleya, Issatchenkia, Kazachstania, Kodameae, Lodderomyces, Pachysolen, Pichia, Satumispora, Starmera, Tetrapisispora, Williopsis, Zygosaccharomyces, Breftanomyces, Kloeckera, Candida  and  Hanseniaspora.    
         [0031]      Saccharomyces:  Genus belonging to the family of Saccharomycetaceae. The genus of  Saccharomyces  can be characterised by e.g. rapid growing colinies which are flat, smooth, glistening or dull, and cream to tannish cream in color. Pseudohyphae may be present; and when present, pseudohyphae are rudimentary. Hyphae are absent. Blastoconidia are 1-celled, globose, and ellipsoid to elongate. Asci contain 1 to 4 ascospores, and do not rupture at maturity. Ascospores often are globose. Nitrate is not utilized. The skilled artisan will be able to determine if a given yeast belongs to the genus of  Saccharomyces,  including  Saccharomyces cerevisiae,  by using standard references such as e.g. Kurtzman, C. P. and J. W. Fell (editors). 1998. The yeasts, a taxonomic study. 4th ed., Elsevier, N.Y.  
         [0032]      Saccharomyces  species: A much preferred species is  Saccharomyces cerevisiae,  optionally in isolated form. Additionally preferred species includes, but is not limited to:  Saccharomyces bamettii, Saccharomyces bayanus, Saccharomyces castellii, Saccharomyces dairenensis, Saccharomyces exiguus, Saccharomyces kluyveri, Saccharomyces paradoxus, Saccharomyces pastorianus, Saccharomyces rosinii, Saccharomyces servazzii, Saccharomyces spencerorum, Saccharomyces transvaalensis,  and  Saccharomyces unisporus. Saccharomyces cerevisiae  deposited with the CBS under accession number CBS 1171 is claimed in connection with the present invention.  
         [0033]      Saccharomyces cerevisiae  synonyms: The following species are listed with the CBS database as synonyms of  Saccharomyces cerevisiae: Candida robusta; Cryptococcus fermentans; Hormiscium cerevisiae; Mycotorula robusta; Saccharomyces aceti; Saccharomyces acidosaccharophillii; Saccharomyces anamensis; Saccharomyces annulatus; Saccharomyces batatae; Saccharomyces beticus; Saccharomyces boulardii; Saccharomyces brasiliensis; Saccharomyces busae asiaticae; Saccharomyces capensis; Saccharomyces carbalali; Saccharomyces carlsbergensis; Saccharomyces carlsbergensis  var.  alcoholophilus; Saccharomyces carlsbergensis  var.  manchuricus; Saccharomyces cerevisiae  Meyen ex E.C. Hansen var.  cerevisiae; Saccharomyces cerevisiae f. pini Saccharomyces cerevisiae  subsp.  orsati; Saccharomyces cerevisiae  subsp.  vetrozensis; Saccharomyces cerevisiae  var.  cratericus; Saccharomyces cerevisiae  var.  ellipsoideus; Saccharomyces cerevisiae  var.  fructuum; Saccharomyces cerevisiae  var.  marchalianus; Saccharomyces cerevisiae  var.  onychophilus; Saccharomyces cerevisiae  var.  pelliculosus; Saccharomyces cerevisiae  var.  pulmonalis; Saccharomyces cerevisiae  var.  turbidans; Saccharomyces cerevisiae agavica sylvestre; Saccharomyces chevalieri  var.  chevalieri; Saccharomyces chevalieri  var.  lindneri; Saccharomyces chodati; Saccharomyces chodati; Saccharomyces cordubensis; Saccharomyces coreanus; Saccharomyces diastaticus; Saccharomyces ellipsoideus  subsp.  alpestris; Saccharomyces ellipsoideus  subsp.  alpinus; Saccharomyces ellipsoideus  subsp.  fulliensis; Saccharomyces ellipsoideus  subsp.  montibensis; Saccharomyces ellipsoideus  subsp.  thermophilus; Saccharomyces ellipsoideus  var.  ellipsoideus; Saccharomyces ellipsoideus  var.  major; Saccharomyces ellipsoideus  var.  umbra; Saccharomyces elongatus; Saccharomyces eryobotryae; Saccharomyces formosensis; Saccharomyces fructuum; Saccharomyces gaditensis; Saccharomyces hienipiensis; Saccharomyces hispalensis; Saccharomyces hispanica; Saccharomyces hutensis; Saccharomyces ilicis; Saccharomyces italicus  var.  italicus; Saccharomyces italicus  var.  melibiosi; Saccharomyces lindneri; Saccharomyces logos; Saccharomyces mangini  var.  casei; Saccharomyces mangini  var.  mangini; Saccharomyces mangini  var.  miso; Saccharomyces marchalianus; Saccharomyces melibiosi; Saccharomyces muentzii; Saccharomyces multisporus; Saccharomyces norbensis; Saccharomyces nutensis; Saccharomyces odessa; Saccharomyces oleaceus; Saccharomyces oleaginosus; Saccharomyces onubensis; Saccharomyces oviformis  var.  bisporus; Saccharomyces oviformis  var.  cheriensis; Saccharomyces oviformis  var.  oviformis; Saccharomyces oxidans; Saccharomyces pastorianus  subsp.  arbignensis; Saccharomyces peka; Saccharomyces prostoserdovii; Saccharomyces pulmonalis; Saccharomyces robustus; Saccharomyces shaoshing; Saccharomyces steineri; Saccharomyces thermantitonum; Saccharomyces tokyo; Saccharomyces turbidans; Saccharomyces valesiacus; Saccharomyces veronae  var.  osloensis; Saccharomyces vordermanii; Saccharomyces wildiersil; Saccharomyces willianus; Saccharomyces yedo; Torula cerevisiae; Torulopsis fermentans;  and  Torulopsis sexta.  Accordingly, the above species are also comprised by the term  Saccharomyces cerevisiae  as used herein. The list is non-limiting and does not exclude that further strains shall be considered synonyms of  Saccharomyces cerevisiae.    
         [0034]     Non- Saccharomyces:  As used herein any of the geni belonging to the family of Saccharomycetaceae excluding the genus of  Saccharomyces.  Preferred examples include, but is not limited to:  Kluyveromyces, Torulaspora, Arxiozyma, Citeromyces, Debaryomyces, Dekkera, Holleya, Issatchenkia, Kazachstania, Kodameae, Lodderomyces, Pachysolen, Pichia, Saturnispora, Starmera, Tetrapisispora, Williopsis,  and  Zygosaccharomyces.    
         [0035]      Torulaspora  species: A much preferred species is  Torulaspora delbrueckii,  optionally in isolated form. Additionally preferred species include, but is not limited to:  T. alkoholi  var  alkoholi, T. alkoholi  var.  azymatica, T. amurcae, T. bailii, T. benedictae, T. bispora, T. californica, T. carsonii, T. castelli, T. cidri, T. coudertii, T. etchellsii, T. eupagyca, T. exigua, T. fermentati, T. florentina, T. formicaria, T. fransiscae, T. globosa, T. hansenii, T. inconspicua, T. kloeckeriana, T. kluyveri, T. lactis a, T. manchuriana, T. melissophila, T. microellipsoides, T. mongolica, T. montana, T. mrakii, T. nilssonii, T. phaffli, T. polymorpha, T. pretoriensis, T. pseudopolymorpha, T. rosei, T. rouxii, T. tamarii, T. vafer, T. vanrijiae,  and  T. yarrowii.  The following  Torulaspora  organisms are claimed in connection with the present invention:  Torulaspora delbrueckii  CBS 1090, CBS 1146, CBS 1148, CBS 1150, CBS 1151, CBS 1152, CBS 1323, CBS 1324, CBS 133, CBS 158, CBS 2733, CBS 2924, CBS CBS 2925, CBS 3003, CBS 3018, CBS 3085, CBS 404, CBS 4510, CBS 4662, CBS 4663, CBS 4664, CBS 4665, CBS 4865, CBS 5448, CBS 5636, CBS 5694, CBS 6104, CBS 6105, CBS 6105.2, CBS 6339, CBS 6518, CBS 6749, CBS 6795, CBS 6871, CBS 6991, CBS 705, CBS 728, CBS 7443, CBS 813, CBS 816, CBS 817, CBS 818, CBS 8247. In one preferred embodiment of the invention,  T. delbrueckii  CBS 3085 is preferred. The genus of  Torulaspora,  including the species of  Torulaspora delbrueckii,  can be characterised by e.g. rapid growing colonies which are round to oval, smooth, and white to cream in color. No hyphae are absent. Asexual reproduction is by budding. Asci contain 1 to 4 ascospores. Ascospores often are often oval and rough. Conjugation occurs between cells and between a cell and its bud. The skilled artisan will be able to determine if a given yeast belongs to the genus of  Torulaspora,  including  Torulaspora delbrueckii,  by using standard references such as e.g. Kurtzman, C. P. and J. W. Fell (editors). 1998. The yeasts, a taxonomic study. 4th ed., Elsevier, N.Y.  
         [0036]      Torulaspora delbrueckii  synonyms: The following species are listed with the CBS database as synonyms of  Torulaspora delbrueckii: Candida colliculosa; Cryptococcus colliculosus; Debaryomyces dekkerae; Debaryomyces delbrueckii; Debaryomyces nilssonii; Debaryomyces rosei; Eutorula colliculosa; Saccharomyces chevalieri  var.  torulosus; Saccharomyces delbrueckii  var.  delbrueckii; Saccharomyces delbrueckii  var.  mongolicus; Saccharomyces fermentati; Saccharomyces florenzanoi; Saccharomyces inconspicuus; Saccharomyces microellipsoides  var.  osmophilus; Saccharomyces nilssonii  var.  nilssonii; Saccharomyces rosei; Saccharomyces saitoanus; Saccharomyces torulosus; Saccharomyces uvarum  var.  inulyticus; Saccharomyces vafer; Schwanniomyces hominis; Torula colliculosa; Torulaspora benedictae; Torulaspora fermentati; Torulaspora inconspicua; Torulaspora mongolica; Torulaspora nilssonii; Torulaspora rosei; Torulaspora vafer; Torulopsis cambresieri; Torulopsis colliculosa; Torulopsis stellata  var.  cambresieri; Torulopsis taboadae; Zygosaccharomyces delbrueckii; Zygosaccharomyces fermentati; Zygosaccharomyces globiformis; Zygosaccharomyces globiformis f. coronata; Zygosaccharomyces globiformis f. typica; Zygosaccharomyces mongolicus; Zymodebaryomyces dekkerae; Zymodebaryomyces delbrueckii;  and  Zymodebaryomyces rosei.  Accordingly, the above species are also comprised by the term  Torulaspora delbrueckii  as used herein. The list is non-limiting and does not exclude that further strains shall be considered synonyms of  Torulaspora delbrueckii.    
         [0037]      Kluyveromyces  species: A much preferred species is  Kluyveromyces thermotolerans,  optionally in isolated form. Additionally preferred species include, but is not limited to:  K.  subgenus  Flabospora, K.  subgenus  Globospora, K. aestuafil, K. africanus, K. bacillisporus, K. blattae, K. bulgaricus, K. cellobiovorus, K. delphensis, K. dobzhanskii, K. drosophilarum, K. fragilis, K. lactis  var,  drosophilarum, K. lactis  var.  lactis, K. lodderae, K. marxianus  var.  bulgaricus, K. marxianus  var.  dobzhanskii, K. marxianus  var.  drosophilarum, K. marxianus  var.  lactis, K. marxianus  var.  marxianus, K. marxianus  var.  vanudenii, K. marxianus  var.  wikenii, K. nonfermentans, K. osmophilus, K. penaeid, K. phaffli, K. phaeseolosporus, K. piceae, K. polysporus, K. sinensis, K. vanudenil, K. veronae, K. waltii, K. wickerhamii, K. wikenii, K. yarrowii.  The following  Kluyveromyces  organisms identified by database accession number are hereby claimed in accordance with the present invention:  Kluyveromyces thermotolerans  CBS 2803 (also available under ATCC 24177; DBVPG 6166;IFO 1674;NCYC 701;NRRL Y-2232;UCD 55-41;VKM Y-533); CBS 7220, CBS 6925, CBS 6433, CBS 4836, CBS 4835, CBS 4834, CBS 4833, CBS 4832. In one preferred embodiment  Kluyveromyces themotolerans  CBS 7220 is preferred. The genus of  Kluyveromyces,  including the species of  Kluyveromyces thermotolerans,  can be characterised by e.g. rapid growing colonies which are round to oval, smooth, and cream in color. Pseudohyphae may be absent. Asexual reproduction is by budding. Asci contain 1 to 4 ascospores. Ascospores often are oval. Conjugation may occur between cells and between a cell and its bud. The skilled artisan will be able to determine if a given yeast belongs to the genus of  Kluyveromyces,  including  Kluyveromyces thermotolerans,  by using standard references such as e.g. Kurtzman, C. P. and J. W. Fell (editors). 1998. The yeasts, a taxonomic study. 4th ed., Elsevier, N.Y.  
         [0038]      Kluyveromyces thermotolerans  synonyms: The following species are listed with the CBS database as synonyms of  Kluyveromyces thermotolerans: Kluyveromyces  Van der Walt,  Kluyveromyces veronae; Kluyveromyces subgenus Fabospora; Candida dattila; Cryptococcus dattilus; Mycotorula dattila; Saccharomyces drosophilae; Saccharomyces thermotolerans; Saccharomyces veronae  var.  veronae; Zygosaccharomyces drosophilae; Zygosaccharomyces thermotolerans; Torula dattila; Torulopsis dattila  var.  dattila; Dekkeromyces  sp.;  Fabospora  sp.;  Zygofabospora thermotolerans;  and  Zygorenospora  sp. Accordingly, the above species are also comprised by the term  Kluyveromyces thermotolerans  as used herein. The list is non-limiting and does not exclude that further strains shall be considered synonyms of  Kluyveromyces thermotolerans.    
         [0039]     Yeast starter culture: Culture comprising at least one yeast species in liquid culture, liquid pressed culture, frozen form or dried form, including freeze dried form and spray/fluid bed dried form. The culture can be packed in vacuum, or under a protected atmosphere such as e.g. nitrogen and the like. The culture is capable of initiating an industrial fermentation process under practical conditions, optionally after being cultivated in a separate starter medium for obtaining a high density culture. The culture can be added to a fermentable carbohydrate source such as e.g. grape juice once or more times during a fermentation process, and it can be added initially and/or later in the fermentation process.  
         [0040]     Mixed yeast starter culture: Yeast starter culture comprising at least one yeast species selected from the genus of  Saccharomyces  and at least one yeast species selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces.    
         [0041]     Taxonomic determinations: The skilled artisan has at his disposal several tools for determining which family, genus and species any particular yeast organism belongs to. For example, recent developments in molecular biology and protein chemistry have provided methods such as DNA restriction fragment length polymorphisms, protein electrophoresis patterns and chromosome fingerprinting. Such techniques have been used for identifying e.g. fermentation-related microorganisms. See, for example, Casey et al., Journal of the American Society of Brewing Chemists, 48(3): 100-106, 1990; Degre et al., American Journal of Enology and Viticulture, 40(4):309-315, 1989; Guillamon et al., Systematic and Applied Microbiology, 19:122-132, 1992; Hoeben et al., Current Genetics, 10:371-379, 1986, Mozina et al., Letters in Applied Microbiology, 24(4):311-315, 1997; Paffetti et al., Research Microbiology, 146:587-594, 1995; Panchal et al., Journal of the Institute of Brewing, 93:325-327, 1987; Querol et al., Systematic and Applied Microbiology, 15:439-446, 1992, Vezinhet et al., Applied Microbiology and Biotechnology, 32:568-571, 1990, and Vezinhet et al., American Journal of Enology and Viticulture, 43(1):83-86, 1992. Polymerase chain reaction (PCR)-based techniques have also been used to detect fermentation-related microorganisms. See, for example, DeBarros Lopes et al., Applied and Environmental Microbiology, 62(12):4514-4520, 1996; Fell, Molecular Marine Biology and Biotechnology, 2(3): 174-180, 1993; Fell, Journal of Industrial Microbiology, 14(6):475-477, 1995; Ibeas et al., Applied and Environmental Microbiology, 62(3):998-1003, 1996; Lavallee et al., American Journal of Enology and Viticulture, 45(1):86-91, 1994; Lieckfeldt et al., Journal of Basic Microbiology, 33(6):413-425, 1993, and Ness et al., J Sci. Food Agric., 62:89-94, 1993. In a different approach, ribosomal genes have been used as molecular probe targets because of their high copy number. Non-transcribed and transcribed spacer sequences associated with ribosomal genes are usually poorly conserved and, thus, are advantageously used as target sequences for the detection of recent evolutionary divergence. Fungral rRNA genes are organized in units. Each unit encodes mature subunits of 18S, 5.8S, and 28S rRNA. The internal transcribed spacer (ITS) region lies between the 18S and 28S rRNA genes and contains two variable non-coding spacers (referred to as ITS1 and ITS2) and the 5.8S rRNA gene (White et al., 1990; In: PCR Protocols; Eds.: Innes et al pages 315-322). In addition, the transcriptional units are separated by non-transcribed spacer sequences (NTSs). The ITS and NTS sequences are particularly suitable molecular probe targets. 
     
    
     FIGURES  
       [0042]     The following figures illustrate the basic physiological properties of  Saccharomyces cerevisiae, Torulaspora delbrueckii  and  Kluyveromyces thermotolerans,  as single strains and mixtures thereof, when used as starter cultures for conversion of grape juice, exemplified but not limited to the grape varieties Pinot noir and Chardonnay. The figures illustrate the physiological limitations of  Torulaspora delbrueckii  and  Kluyveromyces thermotolerans  when used as pure strains, in their ability to convert the grape juice into wine, and the surprising disclosure of this invention when using mixed yeast starter cultures of  Saccharomyces cerevisiae  and  Torulaspora delbrueckii  or  Saccharomyces cerevisiae  and  Kluyveromyces thermotolerans,  or any other combinations thereof.  
         [0043]     Below are provided two different non-limiting examples of grape juice, each one representing an ecosystem, which is subjected to a fermentation according to the invention.  
         [0044]     Pinot noir grape juice has been selected as one example, but not limited to, that is subjected to red wine fermentation conditions according to the invention.  
         [0045]     Chardonnay grape juice has been selected as another example, but not limited to, that is subjected to white wine fermentation conditions according to the invention.  
         [0046]     The figures have been ordered as follows below:  
         [0047]     Pure yeast strain fermentations.  
         [0048]     Pinot noir grape juice (experimental series I)  
         [0049]     I. 1  Saccharomyces cerevisiae  ( FIG. 1 )  
         [0050]     I. 2  Torulaspora delbrueckii  ( FIG. 2 )  
         [0051]     I. 3  Kluyveromyces thermotolerans  ( FIG. 3 )  
         [0052]     Chardonnay grape juice (experimental series II)  
         [0053]     II. 1  Saccharomyces cerevisiae  ( FIG. 4 )  
         [0054]     II. 2  Torulaspora delbrueckii  ( FIG. 5 )  
         [0055]     II. 3  Kluyveromyces thermotolerans  ( FIG. 6 )  
         [0056]     Mixed yeast strain fermentation  
         [0057]     Pinot noir grape juice (experimental series III)  
         [0058]     III. 1  Saccharomyces cerevisiae/Torulaspora delbrueckii  ( FIG. 7 )  
         [0059]     III. 2  Saccharomyces cerevisiae/Kluyveromyces thermotolerans  ( FIG. 8 )  
         [0060]     Chardonnay grape juice (experimental series IV)  
         [0061]     IV. 1  Saccharomyces cerevisiae/Torulaspora delbrueckii  ( FIG. 9 )  
         [0062]     IV. 2  Saccharomyces cerevisiae/Kluyveromyces thermotolerans  ( FIG. 10 ) 
     
    
     EXPERIMENTAL SERIES I  
       [0063]      FIG. 1  illustrates a pure strain alcoholic fermentation by  Saccharomyces cerevisiae  in a Pinot noir grape juice (pasteurised at 140° C./284° F. in 8 seconds and stored in 5 litre sterile plastic containers at 5° C./39.2° F. until use) as a laboratory experiment in 3 litre scale incubated at 28° C./82.4° F.  S. cerevisiae  was prepared by fluid bed drying as generally known in the art and such as detailed in Example 2 below and re-hydrated as generally known in the art and such as detailed in Example 3 below.  
         [0064]     The figure shows that  Saccharomyces cerevisiae  is able, as a pure culture, to convert glucose and fructose into ethanol and glycerol in only 3 days. The development of CO 2  and acetic acid are not shown. After a minor lag phase, a rapid exponential growth was seen before the cell number decreased significantly at the depletion of the carbohydrates.  
         [0065]      FIG. 2  illustrates a pure strain alcoholic fermentation by  Torulaspora delbrueckii  in a Pinot noir grape juice (pasteurised at 140° C./284° F. in 8 seconds and stored in 5 litre sterile plastic containers at 5° C./39.2° F. until use) in a laboratory experiment in 3 litre scale incubated at 28° C./82.4° F.  Torulaspora delbrueckii  was prepared by fluid bed drying as generally known in the art and such as detailed in Example 2 below and re-hydrated as generally known in the art and such as detailed in Example 3 below.  
         [0066]      T. delbrueckii  as a pure culture was not able to convert all the available glucose and fructose and therefore did not complete the alcoholic fermentation during the experiment. The cells only sustained growth for 2 days, before entering the stationary growth phase.  
         [0067]      FIG. 3  illustrates a pure strain alcoholic fermentation by  Kluyveromyces thermotolerans  in a Pinot noir grape juice (pasteurised at 140° C./284° F. in 8 seconds and stored in 5 litre sterile plastic containers at 5° C./39.2° F. until use) in a laboratory experiment in 3 litre scale incubated at 28° C./82.4° F.  Kluyveromyces thermotolerans  was prepared by fluid bed drying as generally known in the art and such as detailed in Example 2 below and re-hydrated as generally known in the art and such as detailed in Example 3 below.  
         [0068]      K. thermotolerans  as a pure culture was not able to convert all the available glucose and fructose and therefore did not complete the alcoholic fermentation during the experiment. The cells sustained growth for 3 days, whereafter the cell number decreased significantly.  
       EXPERIMENTAL SERIES II  
       [0069]      FIG. 4  illustrates a pure strain alcoholic fermentation by  Saccharomyces cerevisiae  in a Chardonnay grape juice (pasteurised at 140° C./284° F. in 8 seconds and stored in 5 litre sterile plastic containers at 5° C./39.2° F. until use) as a laboratory experiment in 1 litre scale incubated at 20° C./68° F.  S. cerevisiae  was prepared by fluid bed drying as generally known in the art and such as detailed in Example 2 below and re-hydrated as generally known in the art and such as detailed in Example 3 below.  
         [0070]     The figure shows that  Saccharomyces cerevisiae  is able, as a pure culture, to convert glucose and fructose into ethanol and glycerol in 9 days. The development of CO 2  and acetic acid are not shown.  
         [0071]      FIG. 5  illustrates a pure strain alcoholic fermentation by  Torulaspora delbrueckii  in Chardonnay grape juice (pasteurised at 140° C./284° F. in 8 seconds and stored in 5 litre sterile plastic containers at 5° C./39.2° F. until use) in a laboratory experiment in 1 litre scale incubated at 20° C./68° F. Torulaspora delbrueckii was prepared by fluid bed drying as generally known in the art and such as detailed in Example 2 below and re-hydrated as generally known in the art and such as detailed in Example 3 below.  
         [0072]      T. delbrueckii  as a pure culture did not complete the alcoholic fermentation during the experiment. The cells grew exponentially for 2 days, before entering the stationary growth phase.  
         [0073]      FIG. 6  illustrates a pure strain alcoholic fermentation by  Kluyveromyces thermotolerans  in a Chardonnay juice (pasteurised at 140° C./284° F. in 8 seconds and stored in 5 litre sterile plastic containers at 5° C./39.2° F. until use) in a laboratory experiment in 1 litre scale incubated at 20° C./68° F.  Kluyveromyces thermotolerans  was prepared by fluid bed drying as generally known in the art and such as detailed in Example 2 below and rehydrated as generally known in the art and such as detailed in Example 3 below.  
         [0074]      K. thermotolerans  as a pure culture did not complete the alcoholic fermentation during the experiment. The cells grew exponentially for 2 days, before entering the stationary growth phase. No death phase appeared in this experiment.  
         [0000]     Conclusion of Single Strain Fermentations (Experimental Series I and II).  
         [0075]      Saccharomyces cerevisiae  was the only strain that was able to convert Pinot noir grape juice and Chardonnay grape juice into wine. Additionally, the experiments show that  Torulaspora delbrueckii  and  Kluyveromyces thermotolerans  are unable to convert the available glucose and fructose in either grape juices, thereby showing an incomplete fermentation which is not desired by wine producer.  
         [0076]     This gives strong indications that the use of  Torulaspora delbrueckii  and  Kluyveromyces thermotolerans  as single strain yeast starter cultures is not desirable in neither white wine nor red wine fermentations, due to the risk of stuck alcoholic fermentation and/or development of off-flavours and off-aromas from the indigeneous yeast and bacteria flora.  
       EXPERIMENTAL SERIES III  
       [0077]      FIG. 7  illustrates an alcoholic fermentation by a mixed culture of  Saccharomyces cerevisia  and  Torulaspora delbrueckii  of Pinot noir grape juice (pasteurised at 140° C./284° F. in 8 seconds and stored in 5 litre sterile plastic containers at 5° C./39.2° F. until use) in a laboratory experiment in 3 litre scale incubated at 28° C./82.4° F.  S. cerevisiae  and  Torulaspora delbrueckii  were prepared by fluid bed drying as generally known in the art and such as detailed in Example 2 below and rehydrated as generally known in the art and such as detailed in Example 3 below.  
         [0078]     The 1:1 mixture of  S. cerevisiae  and  T. delbrueckii  completed the alcoholic fermentation in 3 days. The cell number of  S. cerevisiae  and  T. delbrueckii,  respectively, increased rapidly until day 2, where  S. cerevisiae  entered stationary growth phase until day 3, where the cells entered the death phase. The cells of  T. delbrueckii  were strongly affected by the growth of  S. cerevisiae,  and hardly grew before entering stationary growth phase, and eventually the death phase.  
         [0079]      FIG. 8  illustrates an alcoholic fermentation by a mixed culture of  Saccharomyces cerevisiae  and  Kluyveromyces thermotolerans  of Pinot noir grape juice (pasteurised at 140° C./284° F. in 8 seconds and stored in 5 litre sterile plastic containers at 5° C./39.2° F. until use) in a laboratory experiment in 3 litre scale incubated at 28° C./82.4° F.  Saccharomyces cerevisiae  and  Kluyveromyces thermotolerans  were prepared by fluid bed drying as generally known in the art and such as detailed in Example 2 below and rehydrated as generally known in the art and such as detailed in Example 3 below.  
         [0080]     The 1:1 mixture of  Saccharomyces cerevisiae  and  Kluyveromyces thermotolerans  completed the alcoholic fermentation in 3 days. The cell number of  Saccharomyces cerevisiae  was affected by the presence of  K. thermotolerans,  and grew slower before reaching stationary phase at day 2, and entered the death phase shortly thereafter.  
       EXPERIMENTAL SERIES IV  
       [0081]      FIG. 9  illustrates an alcoholic fermentation by a mixed culture of  Saccharomyces cerevisiae  and  Torulaspora delbrueckii  of Chardonnay grape juice (pasteurised at 140° C./284° F. in 8 seconds and stored in 5 litre sterile plastic containers at 5° C./39.2° F. until use) in a laboratory experiment in 1 litre scale incubated at 20° C./68° F.  Saccharomyces cerevisiae  and  Torulaspora delbrueckii  were prepared by spray/fluid bed drying as generally known in the art and such as detailed in Example 2 below and rehydrated as generally known in the art and such as detailed in Example 3 below.  
         [0082]     The 1:1 mixture of  Saccharomyces cerevisiae  and  T. delbrueckii  completed the alcoholic fermentation in 9 days. The cell number of  Saccharomyces cerevisiae  grew exponentially until day 3, before entering the stationary growth phase. The cells of  T. delbrueckii  grew exponentially until day 2, before rapidly entering the death phase.  
         [0083]      FIG. 10  illustrates an alcoholic fermentation by a mixed culture of  Saccharomyces cerevisiae  and  Kluyveromyces thermotolerans  of Chardonnay grape juice (pasteurised at 140° C./284° F. in 8 seconds and stored in 5 litre sterile plastic containers at 5° C./39.2° F. until use) in a laboratory experiment in 1 litre scale incubated at 20° C./68° F.  Saccharomyces cerevisiae  and  Kluyveromyces thermotolerans  were prepared by spray/fluid bed drying as generally known in the art and such as detailed in Example 2 below and rehydrated as generally known in the art and such as detailed in Example 3 below.  
         [0084]     The 1:1 mixture of  Saccharomyces cerevisiae  and  Kluyveromyces thermotolerans  completed the alcoholic fermentation in 9 days. The cell number of  Saccharomyces cerevisiae  grew exponentially until day 4, before entering the stationary growth phase. The cells of  Kluyveromyces thermotolerans  grew exponentially until day 2, before rapidly entering the death phase.  
         [0000]     Conclusion of Mixed Culture Fermentations (Experimental Series III and IV).  
         [0085]      Saccharomyces cerevisiae  in combination with either  Torulaspora delbrueckii  or  Kluyveromyces thermotolerans  as mixed starter cultures, showed that  Saccharomyces cerevisiae  is the main driving force in the alcohol fermentation. Both  Torulaspora delbrueckii  and  Kluyveromyces thermotolerans  only grew to a limited extent before entering death phase. However, this relatively short period of active growth, is sufficient to modify the aroma and flavour compounds of the finished wine, as shown in tables 1 and 2.  
         [0000]     Tables  
         [0086]     The tables illustrate, but are not limited to, the aroma and flavour compounds produced by a mixture of  Saccharomyces cerevisiae  and  Torulaspora delbrueckii  or a mixture of  Saccharomyces cerevisiae  and  Kluyveromyces thermotolerans.  The tables illustrate the unexpected finding that the use of mixed yeast starter cultures of  Saccharomyces cerevisiae  and  Torulaspora delbrueckii  or of  Saccharomyces cerevisiae  and  Kluyveromyces thermotolerans  or any combinations thereof gives rise to the development of enhanced aroma and flavour profiles, which are different from those resulting from a pure strain fermentation of  S. cerevisiae.  In addition, the tables show aroma and flavour differences between the use of  Saccharomyces/Torulaspora delbrueckii  and  Saccharomyces/Kluyveromyces thermotolerans.   
                           TABLE 1                                   Chemical compound               (GC-olfactometry)   NIF detection                           3-methyl butyl acetate   Fruit           Ethyl hexanoat   Fruit, peach, pineapple           Hexyl acetate   Flowers, mild fruit           Cis-3-hexenyl acetate   Vitamin pill           n-octyl acetate   Sweet fruit, citrus                      
 
         [0087]     Table 1 illustrates some of the chemical aroma components produced during alcoholic fermentation of Chardonnay grape juice (pasteurised at 140° C./284° F. in 8 seconds and stored in 5 litre sterile plastic containers at 5° C./39.2° F. until use) by a mixed culture of  Saccharomyces cerevisiae  and  Torulaspora delbrueckii,  not related to the grape juice nor a pure strain alcoholic fermentation by  Saccharomyces cerevisiae  under the same conditions. The experiment was conducted in 20 litre scale incubated at 20° C./68° F. The samples were analysed by GC-mass spectrometry (Williams, D. H. &amp; Fleming, I., 1995) and NIF (Pollien et al., 1997)  
         [0088]     Table 2 illustrates some of the chemical aroma components produced during alcoholic fermentation of Chardonnay grape juice by a mixed culture of  Saccharomyces cerevisiae  and  Kluyveromyces thermotolerans  not related to the grape juice nor a pure strain alcoholic fermentation by  Saccharomyces cerevisiae  under the same conditions. The experiment was conducted in 20 litre scale incubated at 20° C./68° F.  
                           TABLE 2                                   Chemical compound               (GC-olfactometry)   NIF detection                           3-methyl butyl acetate   Fruit           Cis-3-hexenyl acetate   Vitamin pill           n-octyl acetate   Sweet fruit, citrus           Methyl octanoat   Hand soap, paper commercials           2-methylbutyl octanoat   Citrus, fruit, flowers, sweet           ethyl nonanoat   Leather           ethyl decanoat   Hop, bad vitamin pill           Propyl decanoat   Citrus, Easter daffodil, pleasant                      
 
         [0089]     A comparison of data from tables 1 and 2 strongly indicates that there are both similarities because of the presence of some of the same compounds, and differences in aroma components as a result of fermentation with  T. delbrueckii/Saccharomyces cerevisiae  compared to  K. thermotolerans/Saccharomyces cerevisiae.    
       DETAILED DESCRIPTION  
       [0000]     Composition Comprising at Least One Non- Saccharomyces  Yeast Organism  
         [0090]     When the invention according to one aspect pertains to a composition comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces,  the at least one first yeast organism is in one embodiment selected from the genus of  Torulaspora.    
         [0091]     The  Torulaspora  organism is preferably selected from the group consisting of  T. delbrueckii, T. alcoholi  var  alcoholi, T. alcoholi  var.  azymatica, T. amurcae, T. baili, T. benedictae, T. bispora, T. californica, T. carsonii, T. castellii, T. cidri, T. coudertii, T. etchellsii, T. eupagyca, T. exigua, T. fermentati, T. florentina, T. formicaria, T. fransiscae, T. globosa, T. hansenii, T. inconspicua, T. kloeckeriana, T. kluyveri, T. lactis a, T. manchuriana, T. melissophila, T. microellipsoides, T. mongolica, T. montana, T. mrakii, T. nilssonii, T. phaffii, T. polymorpha, T. pretoriensis, T. pseudopolymorpha, T. rosei, T. rouxii, T. tamarii, T. vafer, T. vanrijiae, T. yarrowii,  including any combination thereof.  
         [0092]     In another embodiment, the at least one first yeast organism is in one embodiment selected from the genus of  Kluyveromyces.  The  Kluyveromyces  organism is preferably selected from the group consisting of  K. thermotolerans, K.  subgenus  Flabospora, K.  subgenus  Globospora, K. aestuarii, K. africanus, K. bacillisporus, K. blattae, K. bulgaricus, K. cellobiovorus, K. delphensis, K. dobzhanskii, K. drosophilarum, K. fragilis, K. lactis  var,  drosophilarum, K. lactis  var.  lactis, K. lodderae, K. marxianus  var.  bulgaricus, K. marxianus  var.  dobzhanskii, K. marxianus  var.  drosophilarum, K. marxianus  var.  lactis, K. marxianus  var.  marxianus, K. marxianus  var.  vanudenii, K. marxianus  var.  wikenji, K. nonfermentans, K. osmophilus, K. penaeid, K. phaffii, K. phaeseolosporus, K. piceae, K. polysporus, K. sinensis, K. vanudenii, K. veronae, K. waltii, K. wickerhamii, K. wikenii,  and  K. yarrowii,  including any combination thereof.  
         [0093]     The composition is preferably freeze dried or spray/fluid bed dried.  
         [0000]     Composition Comprising at Least One First Non- Saccharomyces  Yeast Organism and at Least One Second Non- Saccharomyces  Yeast Organism  
         [0094]     When the invention according to a second aspect pertains to a composition comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one second yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces,  the at least one first and second yeast organism is in one embodiment selected from the genus of  Torulaspora  and from the genus of  Kluyveromyces,  respectively.  
         [0095]     The  Torulaspora  organism is preferably selected from the group consisting of  T. delbrueckii, T. alcoholi  var  alcoholi, T. alkoholi  var.  azymatica, T. amurcae, T. bailii, T. benedictae, T. bispora, T. californica, T. carsonii, T. castellii, T. cidri, T. coudertii, T. etchellsii, T. eupagyca, T. exigua, T. fermentati, T. florentina, T. formicaria, T. fransiscae, T. globosa, T. hansenii, T. inconspicua, T. kloeckeriana, T. kluyveri, T. lactis a, T. manchuriana, T. melissophila, T. microellipsoides, T. mongolica, T. montana, T. mrakii, T. nilssonii, T. phaffii, T. polymorpha, T. pretoriensis, T. pseudopolymorpha, T. rosei, T. rouxii, T. tamarii, T. vafer, T. vanrijiae, T. yarrowii,  including any combination thereof.  
         [0096]     In combination with any of the above  Torulaspora  organisms, the  Kluyveromyces  organism is preferably selected from the group consisting of  K. thermotolerans, K.  subgenus  Flabospora, K.  subgenus  Globospora, K. aestuarii, K. africanus, K. bacillisporus, K. blattae, K. bulgaricus, K. cellobiovorus, K. delphensis, K. dobzhanskii, K. drosophilarum, K. fragilis, K. lactis  var,  drosophilarum, K. lactis  var.  lactis, K. lodderae, K. marxianus  var.  bulgaricus, K. marxianus  var.  dobzhanskii, K. marxianus  var.  drosophilarum, K. marxianus  var.  lactis, K. marxianus  var.  marxianus, K. marxianus  var.  vanudenii, K. marxianus  var.  wikenii, K. nonfermentans, K. osmophilus, K. penaeid, K. phaffli, K. phaeseolosporus, K. piceae, K. polysporus, K. sinensis, K. vanudenii, K. veronae, K. waltii, K. wickerhamii, K. wikenii, K. yarrowii,  including any combination thereof.  
         [0097]     In one particularly preferred embodiment there is provided a composition, wherein the  Torulaspora  organism is  Torulaspora delbrueckii  and wherein the  Kluyveromyces  organism is  Kluyveromyces thermotolerans.    
         [0098]     The composition is preferably freeze dried or spray/fluid bed dried.  
         [0000]     Composition Comprising One or More Non- Saccharomyces  Yeast Organisms and at Least One Further  Saccharomyces  Yeast Organism  
         [0099]     When the invention according to a third aspect relates to either a composition comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one further yeast organism selected from the genus of  Saccharomyces,  or relates to a composition comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one second yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one further yeast organism selected from the genus of  Saccharomyces,  the first and/or second (non- Saccharomyces ) yeast organisms can be any of the (combinations of) organisms described herein above, whereas the at least one further yeast organism is preferably selected from the group consisting of  Saccharomyces cerevisiae, Saccharomyces barnettii, Saccharomyces bayanus, Saccharomyces castellii, Saccharomyces dairenensis, Saccharomyces exiguus, Saccharomyces kluyveri, Saccharomyces paradoxus, Saccharomyces pastorianus, Saccharomyces rosinii, Saccharomyces servazzii, Saccharomyces spencerorum, Saccharomyces transvaalensis,  and  Saccharomyces Saccharomyces transvaalensis,  and  Saccharomyces unisporus,  including any combination thereof.  
         [0100]      Saccharomyces cerevisiae  is preferred, optionally in combination with  Torulaspora delbrueckii  and  Kluyveromyces thermotolerans.    
         [0101]     The composition is preferably freeze dried or spray/fluid bed dried.  
         [0000]     Colony Forming Units and Ratios Between Individual Yeast Organisms  
         [0102]     When the above compositions are to be used as starter cultures it is desirable to have a number of colony forming units per gram (dry weight) starter culture which is preferably more than about 10 8  CFU for the at least one further yeast organism, such as more than about 10 9  CFU, for example more than about 10 10  CFU, such as more than about 2×10 10  CFU.  
         [0103]     Likewise, it is desirable to have a number of colony forming units per gram (dry weight) starter culture which is preferably more than about 10 7  CFU for the at least one first yeast organism and the same number of CFU for the at least one second yeast organism, when present, such as more than about 10 8  CFU, for example more than about 10 9  CFU, such as more than about 2×10 9  CFU.  
         [0104]     When the composition comprises the at least one first yeast organism and the at least one second yeast organism, the ratio R between i) the number (CFU) of the at least one first yeast organism cells, and ii) the number (CFU) of the at least one second yeast organism cells in the composition, is preferably in the range of from more than 0.1 to preferably less than 100, such as in the range of from 0.1 to 0.3, for example in the range of from 0.2 to 0.3, such as in the range of from 0.3 to 0.5, for example in the range of from 0.4 to 0.5, such as in the range of from 0.5 to 0.7, for example in the range of from 0.6 to 0.7, such as in the range of from 0.7 to 0.9, for example in the range of from 0.8 to 0.9, such as in the range of from 0.9 to 1.0 and such as about 1.0, for example in the range of from 1.0 to 1.1, such as in the range of from 1.1 to 1.3, for example in the range of from 1.2 to 1.3, such as in the range of from 1.3 to 1.5, for example in the range of from 1.4 to 1.5, such as in the range of from 1.6 to 1.8, for example in the range of from 1.7 to 1.8, such as in the range of from 1.8 to 2.0, for example in the range of from 1.9 to 2.0, such as in the range of from 2.0 to 2.4, for example in the range of from 2.2 to 2.4, such as in the range of from 2.4 to 3.0, for example in the range of from 2.8 to 3.0, such as in the range of from 3.0 to 4.0, for example in the range of from 3.5 to 4.0, such as in the range of from 4.0 to 5.0, for example in the range of from 4.5 to 5.0, such as in the range of from 5.0 to 7.0, for example in the range of from 6.0 to 7.0, such as in the range of from 7.0 to 9.0, for example in the range of from 8.0 to 9.0, such as in the range of from 9.0 to 12, for example in the range of from 10 to 12, such as in the range of from 12 to 16, for example in the range of from 14 to 16, such as in the range of from 16 to 20, for example in the range of from 18 to 20, such as in the range of from 20 to 30, for example in the range of from 25 to 30, such as in the range of from 30 to 50, for example in the range of from 40 to 50, and such as in the range of from 50 to 100.  
         [0105]     When the composition comprises the at least one first yeast organism and the at least one further yeast organism the ratio R between i) the number (CFU) of the at least one first yeast organism cells, and ii) the number (CFU) of the at least one further yeast organism cells, is preferably in the range of from more than 0.1 to preferably less than 100, such as in the range of from 0.1 to 0.3, for example in the range of from 0.2 to 0.3, such as in the range of from 0.3 to 0.5, for example in the range of from 0.4 to 0.5, such as in the range of from 0.5 to 0.8, for example in the range of from 0.6 to 0.8, such as in the range of from 0.7 to 0.9, for example in the range of from 0.8 to 0.9, such as in the range of from 0.9 to 1.0 and such as about 1.0, for example in the range of from 1.0 to 1.1, such as in the range of from 1.1 to 1.3, for example in the range of from 1.2 to 1.3, such as in the range of from 1.3 to 1.5, for example in the range of from 1.4 to 1.5, such as in the range of from 1.6 to 1.8, for example in the range of from 1.7 to 1.8, such as in the range of from 1.8 to 2.0, for example in the range of from 1.9 to 2.0, such as in the range of from 2.0 to 2.4, for example in the range of from 2.2 to 2.4, such as in the range of from 2.4 to 3.0, for example in the range of from 2.8 to 3.0, such as in the range of from 3.0 to 4.0, for example in the range of from 3.5 to 4.0, such as in the range of from 4.0 to 5.0, for example in the range of from 4.5 to 5.0, such as in the range of from 5.0 to 7.0, for example in the range of from 6.0 to 7.0, such as in the range of from 7.0 to 9.0, for example in the range of from 8.0 to 9.0, such as in the range of from 9.0 to 12, for example in the range of from 10 to 12, such as in the range of from 12 to 16, for example in the range of from 14 to 16, such as in the range of from 16 to 20, for example in the range of from 18 to 20, such as in the range of from 20 to 30, for example in the range of from 25 to 30, such as in the range of from 30 to 50, for example in the range of from 40 to 50, and such as in the range of from 50 to 100.  
         [0106]     When the composition comprises the at least one first and second yeast organism and the at least one further yeast organism, the ratio R between i) the number (CFU) of cells of the at least one first yeast organism and the at least one second yeast organism, and ii) the number (CFU) of the at least one further yeast organism cells, is preferably in the range of from more than 0.1 to preferably less than 100, such as in the range of from 0.1 to 0.3, for example in the range of from 0.2 to 0.3, such as in the range of from 0.3 to 0.5, for example in the range of from 0.4 to 0.5, such as in the range of from 0.5 to 0.8, for example in the range of from 0.6 to 0.8, such as in the range of from 0.7 to 0.9, for example in the range of from 0.8 to 0.9, such as in the range of from 0.9 to 1.0 and such as about 1.0, for example in the range of from 1.0 to 1.1, such as in the range of from 1.1 to 1.3, for example in the range of from 1.2 to 1.3, such as in the range of from 1.3 to 1.5, for example in the range of from 1.4 to 1.5, such as in the range of from 1.6 to 1.8, for example in the range of from 1.7 to 1.8, such as in the range of from 1.8 to 2.0, for example in the range of from 1.9 to 2.0, such as in the range of from 2.0 to 2.4, for example in the range of from 2.2 to 2.4, such as in the range of from 2.4 to 3.0, for example in the range of from 2.8 to 3.0, such as in the range of from 3.0 to 4.0, for example in the range of from 3.5 to 4.0, such as in the range of from 4.0 to 5.0, for example in the range of from 4.5 to 5.0, such as in the range of from 5.0 to 7.0, for example in the range of from 6.0 to 7.0, such as in the range of from 7.0 to 9.0, for example in the range of from 8.0 to 9.0, such as in the range of from 9.0 to 12, for example in the range of from 10 to 12, such as in the range of from 12 to 16, for example in the range of from 14 to 16, such as in the range of from 16 to 20, for example in the range of from 18 to 20, such as in the range of from 20 to 30, for example in the range of from 25 to 30, such as in the range of from 30 to 50, for example in the range of from 40 to 50, and such as in the range of from 50 to 100.  
         [0107]     When the composition or yeast starter culture comprises a)  Saccharomyces  sp., including  Saccharomyces cerevisiae,  b)  Torulaspora  sp., including  Torulaspora delbrueckii,  and c)  Kluyveromyces  sp., including  Kuyveromyces thermotolerans,  the ratios (in colony forming units, CFU) among the aforementioned species a):b):c) is in one embodiment preferably about 1:1:1, such as about 1:1:2; for example 1:1:3, such as about 1:1:4; for example 1:1:5; such as about 1:2:1, for example 1:2:2; such as about 1:2:3, for example 1:2:4; such as about 1:2:5, for example 1:3:1; such as about 1:3:2, for example 1:3:3; such as about 1:3:4, for example 1:3:5; such as about 1:4:1, for example 1:4:2; such as about 1:4:3, for example 1:4:4; such as about 1:4:5, for example 1:5:1; such as about 1:5:2, for example 1:5:3; such as about 1:5:4, for example 1:5:5 and such as about 5:1:1 and such as about 5:1:2, for example 5:1:3; such as about 5:1:4, for example 5:1:5; such as about 5:2:1, for example 5:2:2; such as about 5:2:3, for example 5:2:4; such as about 5:2:5, for example 5:3:1; such as about 5:3:2, for example 5:3:3; such as about 5:3:4, for example 5:3:5; such as about 5:4:1, for example 5:4:2; such as about 5:4:3, for example 5:4:4; such as about 5:4:5, for example 5:5:1; such as about 5:5:2, for example 5:5:3; such as about 5:5:4, for example 10:1:1; such as about 10:1:2, for example 10:1:3; such as about 10:1:4, for example 10:1:5; such as about 10:2:1, such as about 10:2:3 and such as about 10:2:5, for example 10:3:1; such as about 10:3:2, for example 10:3:3; such as about 10:3:4, for example 10:3:5; such as about 10:4:1 and such as about 10:4:3 and such as about 10:4:5, for example 10:5:1; such as about 10:5:2, for example 10:5:3; such as about 10:5:4, for example 20:1:1; such as about 20:1:2, for example 20:1:3; such as about 20:1:4, for example 20:1:5; such as about 20:2:1 and such as about 20:2:3 and such as about 20:2:5, for example 20:3:1; such as about 20:3:2, for example 20:3:3; such as about 20:3:4, for example 20:3:5; such as about 20:4:1 and such as 20:4:3 and such as 20:4:5, for example 20:5:1; such as about 20:5:2, for example 20:5:3; such as about 20:5:4, for example 20:5:5 and for example 30:1:1; such as 30:1:2; for example 30:1:3; such as 30:1:4; for example 30:1:5; such as 30:2:1; for example 30:2:2; such as 30:2:3; for example 30:2:4; such as 30:2:5; for example 30:3:1; such as 30:3:2; such as about 30:3:4, for example 30:3:5; such as about 30:4:1, for example 30:4:2; such as about 30:4:3, for example 30:4:4; such as about 30:4:5, for example 30:5:1; such as about 30:5:2, for example 30:5:3; such as about 30:5:4, for example 30:5:5 and for example 40:1:1; such as about 40:1:2, for example 40:1:3; such as about 40:1:4, for example 40:1:5; such as about 40:2:1 and such as 40:2:3 and such as 40:2:5, for example 40:3:1; such as about 40:3:2, for example 40:3:3; such as about 40:3:4, for example 40:3:5; such as about 40:4:1 and such as about 40:4:3 and such as about 40:4:5, for example 40:5:1; such as about 40:5:2, for example 40:5:3; such as about 40:5:4, for example 40:5:5, such as about 50:1:1 and such as about 50:1:2, for example 50:1:3; such as about 50:1:4, for example 50:1:5; such as about 50:2:1, for example 50:2:2; such as about 50:2:3, for example 50:2:4; such as about 50:2:5, for example 50:3:1; such as about 50:3:2, for example 50:3:3; such as about 50:3:4, for example 50:3:5; such as about 50:4:1, for example 50:4:2; such as about 50:4:3, for example 50:4:4; such as about 50:4:5, for example 50:5:1; such as about 50:5:2, for example 50:5:3; such as about 50:5:4, for example 50:5:5.  
         [0000]     Methods Relating to the Composition of the Invention  
         [0108]     It is understood that the invention also relates to a method for fermenting at least one fermentable carbohydrate source in an aqueous composition. This method comprises the step of contacting the aqueous composition with the composition of the invention under conditions allowing said fermentation to occur, and in this case the carbohydrate source preferably comprises at least one pentose or at least one hexose, including any monomer, dimer, oligomer or polymer thereof. Preferred fermentable carbohydrate sources are e.g. fructose, glucose, galactose, maltose, sucrose, trehalose, melibiose, starch, raffinose, including any combination thereof. However, other carbohydrate sources can also be used.  
         [0109]     When the aqueous composition comprises grape juice, the carbohydrate sources can be obtained from white grapes or red grapes, or both. The white grapes can be selected from the group of grapes consisting of Aligote, Chardonnay, Chenin Blanc, Columbard, Folle Blanche, Gewurztraminer, Groner Veltliner, Malvasia, Marsanne, Melon de Bourgogne, Muller-Thurgau, Muscadelle, Muscat, Palomino, Pedro Ximenez, Pinot Blanc, Pinot Gris/Pinot Grigio, Riesling, Rousanne, Sauvignon Blanc/Fume Blanc, Scheurebe, Semillion, Sylvaner, Trebbiano, Ugni Blanc, Verdicchio, Viognier, including any combination thereof.  
         [0110]     The red grapes can be selected from the group of grapes consisting of Barbera, Brunello, Cabernet Franc, Cabernet Sauvignon, Carignana, Carmenere, Cinsault, Dolcetto, Dornfelder, Gamay, Grenache, Grignolino, Malbec, Merlot, Montepulciano, Mourvedre/Mataro, Nebbiolo, Petit Sirah, Petit Verdot, Pinotage, Pinot Meunier, Pinot Noir, Primitivo, Rondo, Sangiovese, Syrah/Shiraz, Tempranillo, Tinta Barroca, Tinta Cao, Toriga Francesa, Touriga Nacional, Tinta Roriz, Zinfandel, including any combination thereof.  
         [0111]     As many wine districts have their own preferred combination of grapes, such combinations are also forming part of the invention. In some cases, wine can only be produced within a particular district when strict rules for grape combinations and quantities are adhered to. The skilled person will be familiar with such rules.  
         [0112]     When the invention relates to a method for producing an edible or drinkable product, said method comprising the step of adding the composition according to the invention to a precursor composition of said product, fermenting said precursor composition and producing said edible or drinkable product, the product is in one embodiment an alcoholic beverage including wine, such as red wine, rose wine, blush wine, white wine, sparkling wine, and fruit wine. Further examples of alcoholic beverages comprise cider and beer. Examples of edible products comprise bread. Further examples of products are milk and soy.  
         [0113]     In another embodiment, the invention relates to a method for flavouring an edible or drinkable product, said method comprising the step of adding the composition according to the invention to a precursor composition of said product, fermenting said precursor composition and flavouring said edible or drinkable product, preferably an alcoholic beverage such as e.g. wine, for example red wine, rose wine, blush wine, white wine, sparkling wine, and fruit wine, as well as cider and beer. The flavour can also be added to e.g. bread, milk and soy.  
         [0114]     In another preferred embodiment, the invention relates to a method for preparing a dried composition according to the invention, said method comprising the steps of i) providing at least one first yeast organism, and ii) drying said at least one first yeast organism to produce a dried composition, preferably a freeze dried composition or a spray/fluid bed dried composition.  
         [0115]     The method in one preferred embodiment comprises the steps of i) providing at least one first yeast organism selected from the genus of  Torulaspora  or the genus of  Kluyveromyces,  and ii) drying said composition, preferably by freeze drying or by spray/fluid bed drying. The first yeast organism is preferably  Torulaspora delbrueckii  or  Kluyveromyces thermotolerans.    
         [0116]     The method in another preferred embodiment comprises the steps of i) providing a composition comprising at least one first yeast organism selected from the genus of  Torulaspora  and at least one second yeast species selected from the genus of  Kluyveromyces,  and ii) drying said composition, preferably by freeze drying or by spray/fluid bed drying. The first yeast organism is preferably  Torulaspora delbrueckii  and the second yeast organism is preferably  Kluyveromyces thermotolerans.    
         [0117]     The method in yet another preferred embodiment comprises the steps of i) providing a composition comprising at least one first yeast organism selected from the genus of  Torulaspora  or the genus of  Kluyveromyces,  and at least one further yeast organism selected from the genus of  Saccharomyces,  and ii) drying said composition, preferably by freeze drying or by spray/fluid bed drying. The first yeast organism is preferably  Torulaspora delbrueckii  or  Kluyveromyces thermotolerans,  and the at least one further yeast organism is preferably  Saccharomyces cerevisiae.    
         [0118]     The method in an even further preferred embodiment comprises the steps of i) providing at least one first yeast organism selected from the genus of  Torulaspora  and at least one second yeast organism selected from the genus of  Kluyveromyces  and at least one further yeast organism selected from the genus of  Saccharomyces,  and ii) drying said composition, preferably by freeze drying or by spray/fluid bed drying. The first yeast organism is preferably  Torulaspora delbrueckii,  the second yeast organism is preferably  Kluyveromyces thermotolerans.  and the further yeast organism is preferably  Saccharomyces cerevisiae.    
         [0119]     In further embodiments there are provided the use of a composition according to the invention for the manufacture of a yeast starter culture for fermenting grape juice, including grape juice obtained from white grapes such as e.g. Aligote, Chardonnay, Chenin Blanc, Columbard, Folle Blanche, Gewurztraminer, Groner Veltliner, Malvasia, Marsanne, Melon de Bourgogne, Muller-Thurgau, Muscadelle, Muscat, Palomino, Pedro Ximenez, Pinot Blanc, Pinot Gris/Pinot Grigio, Riesling, Rousanne, Sauvignon Blanc/Fume Blanc, Scheurebe, Semillion, Sylvaner, Trebbiano, Ugni Blanc, Verdicchio, Viognier, including any combination thereof, as well as grape juice obtained from red grapes such as e.g. Barbera, Brunello, Cabernet Franc, Cabernet Sauvignon, Carignana, Carmenere, Cinsault, Dolcetto, Gamay, Grenache, Grignolino, Malbec, Merlot, Montepulciano, Mourvedre/Mataro, Nebbiolo, Petit Sirah, Petit Verdot, Pinotage, Pinot Meunier, Pinot Noir, Sangiovese, Syrah/Shiraz, Tempranillo, Tinta Barroca, Tinta Cao, Toriga Francesa, Touriga Nacional, Tinta Roriz, Zinfandel, including any combination thereof.  
         [0120]     The invention also relates to the use of a composition according to the invention for producing an edible or drinkable product. In yet another embodiment there is provded the use of a composition according to the invention for flavouring an edible or drinkable product. The products include alcoholic beverages, including wine such as red wine, rose wine, blush wine, white wine, sparkling wine, fruit wine, as well as cider and beer. Additional products capable of being produced by the invention are bread, milk and soy.  
         [0121]     There is also provided the use of at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  for the preparation of a yeast starter culture for fermenting grape juice and flavouring an edible or drinkable product resulting from said fermentation. The starter culture is preferably dried such as freeze dried or /fluid bed dried. The first yeast organism is preferably selected from the genus of  Torulaspora,  such as  Torulaspora delbrueckii.  However, the first species can also be selected from the genus of  Kluyveromyces,  including  Kluyveromyces thermotolerans.    
         [0122]     The starter culture is preferably dried by freeze drying or by spray/fluid bed drying when the present invention is used to provide a composition comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one second yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  for the preparation of a yeast starter culture for fermenting grape juice and flavouring an edible or drinkable product resulting from said fermentation. The first yeast organism is preferably selected from the genus of  Torulaspora,  including  Torulaspora delbrueckii,  whereas the second yeast organism is preferably selected from the genus of  Kluyveromyces,  including  Kluyveromyces thermotolerans.    
         [0123]     The starter culture is preferably dried by freeze drying or by spray/fluid bed drying when the present invention is used to provide a composition comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one further yeast organism selected from the genus of  Saccharomyces  for the preparation of a yeast starter culture for fermenting grape juice and flavouring an edible or drinkable product resulting from said fermentation. The first yeast organism is preferably selected from the genus of  Torulaspora,  such as  Torulaspora delbrueckii.  However, the first species can also be selected from the genus of  Kluyveromyces,  including  Kluyveromyces thermotolerans.  The further yeast organism is preferably  Saccharomyces cerevisiae.    
         [0124]     In yet another embodiment there is provided by the present invention the use of a composition comprising at least one first yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one second yeast organism selected from the family of Saccharomycetaceae excluding the genus of  Saccharomyces  and at least one further yeast organism selected from the genus of  Saccharomyces  for the preparation of a yeast starter culture for fermenting grape juice and flavouring an edible or drinkable product resulting from said fermentation.  
         [0125]     The prepared starter culture is preferably dried, such as freeze dried or spray/fluid bed dried. The first yeast organism is preferably selected from the genus of  Torulaspora,  including  Torulaspora delbrueckii,  and the second yeast organism is preferably selected from the genus of  Kluyveromyces,  including  Kluyveromyces thermotolerans.  The further yeast organism is preferably  Saccharomyces cerevisiae.    
       EXAMPLES  
       [0126]     The examples illustrate preferred embodiments of the invention without limiting the invention to such embodiments. In this context starter cultures is to be understood as inoculation material of the appropriate yeast strain for large-scale production.  
       Example 1  
       [0000]     Preparation of Starter Culture for Wine Fermentation by Means of Freeze Drying  
         [0127]     The following examples illustrate the production and freeze drying of starter culture consisting of either  Saccharomyces cerevisiae,  or  Torulaspora delbrueckii,  or  Kluyveromyces thermiotolerans.    
         [0128]     Preparation of YPG growth medium. A mixture of 10 g/L D-Glucose, 5 g/L yeast extract, and 10 g/L peptone (YPG) adjusted to pH 5.5 adding 1N HCl before autoclavation at 121° C./249.8° F. for 20 minutes.  
         [0129]     Preparation of YPG agar. A mixture of 10 g/L D-Glucose, 5 g/L yeast extract, 20 g/L agar, and 10 g/L peptone (YPG) adjusted to pH 5.5 adding 1 N HCl before autoclavation at 121° C./249.8° F. for 20 minutes. The liquid agar solution is dispersed in sterile petri dishes stored at 5° C. until use.  
         [0130]     Preparation of inoculation material to be used as starter culture for large-scale production. This procedure is made up of 3 steps, before entering into a large scale tank.  
         [0131]     Step 1. A pure culture ampoule containing the appropriate yeast species kept at −80° C./−112° F. is dispersed on an YPG agar plate and incubated for 48 hours at 30° C./86° F.  
         [0132]     Step 2. From this YPG agar plate, a single colony of the appropriate yeast species is picked, and transferred to an YPG medium. The medium is incubated for 48 hours at 30° C./86° F. under shaking (200 rpm).  
         [0133]     Step 3. This yeast solution is used for preparing a 1% starter culture in semi-large scale to fit an starter culture of 1% in the final large-scale. The 1% starter culture is prepared by diluting in a ratio of 1 to 100 the starter culture with the media used for the large scale production.  
         [0000]     Large Scale Production  
         [0134]     1% inoculation material prepared as described above, is inoculated into production tank containing YPG growth medium. Glucose being the carbohydrate source can be substituted by e.g. molasses or starch. The cells are incubated at 30° C./86° F. while being stirred at 200 rpm. The carbohydrate source is pumped into a tank while correlating with monitoring the optical density and air is pumped into tank while correlating with the ethanol production, i.e. if it is possible to measure the presence of any ethanol, more oxygen is to be pumped into the tank.  
         [0135]     Inlets of air flow and carbohydrate are stopped after 16-18 hours. Cells are ready to be harvested after 24 hours.  
         [0136]     The cells are harvested and concentrated approximately 15 times by centrifugation (e.g. 4000 rpm, 10 minutes). 1% glycerol is added as cryoprotective agent.  
         [0000]     Freeze Drying  
         [0137]     The concentrated cells are poured onto sterile trays and placed in a freeze dryer under vacuum. Water is evaporated from the cells monitored by weight loss during the drying. When the weight becomes constant, the product is dry, and is distributed to packing of sterile aluminium bags in various sizes as follows.  
         [0138]     Freeze dried products of  Saccharomyces cerevisiae  and  T. delbrueckii  are mixed in a desired ratio as described on pp, in package sizes of e.g. 500 g, 1 kg, 5 kg, 25 kg. Freeze dried products of  Saccharomyces cerevisiae  and  K thermotolerans  are mixed in a desired ratio as described, in package sizes of e.g. 500 g, 1 kg, 5 kg, 25 kg.  
         [0139]     Freeze dried products of  Saccharomyces cerevisiae, T. delbrueckii  and  K. thermotolerans  are mixed in a desired ratio as described, in package sizes of e.g. 500 g, 1 kg, 5 kg, 25 kg.  
       Example 2  
       [0000]     Preparation of Starter Culture by Means of Spray/Fluid Bed Drying  
         [0140]     The following example illustrates the production and spray/fluid bed drying of  Saccharomyces cerevisiae, Torulaspora delbrueckii,  and  Kluyveromyces thermotolerans.    
         [0141]     Preparation of YPG growth medium. A mixture of 10 g/L D-Glucose, 5 g/L yeast extract, and 10 g/L peptone (YPG) adjusted to pH 5.5 adding 1 N HCl before autoclavation at 121° C./249.8° F. for 20 minutes.  
         [0142]     Preparation of YPG agar. A mixture of 10 g/L D-Glucose, 5 g/L yeast extract, 20 g/L agar, and 10 g/L peptone (YPG) adjusted to pH 5.5 adding 1 N HCl before autoclavation at 121° C./249.8° F. for 20 minutes. The liquid agar solution is dispersed in sterile petri dishes stored at 5° C. until use.  
         [0143]     Preparation of inoculation material to be used as starter culture for large-scale production. This procedure is made up of 3 steps, before entering into a large scale tank.  
         [0144]     Step 1. A pure culture ampoule containing the appropriate yeast species kept at −80° C./−112° F. is dispersed on an YPG agar plate an incubated for 48 hours at 30° C./86° F.  
         [0145]     Step 2. From this YPG agar plate, a single colony of the appropriate yeast species is picked, and transferred to an YPG medium. The medium is incubated for 48 hours at 30° C./86° F. under shaking (200 rpm).  
         [0146]     Step 3. This yeast solution is used for preparing a 1% starter culture in semi-large scale to fit an starter culture of 1% in the final large-scale. The 1% starter culture is prepared by diluting in a ratio of 1 to 100 the starter culture with the media used for the large scale production.  
         [0000]     Large Scale Production  
         [0147]     1% inoculation material prepared as described above, is inoculated into production tank containing YPG growth medium. Glucose being the carbohydrate source can be substituted by e.g. molasses or starch. The cells are incubated at 30° C./86° F. while being stirred at 200 rpm. The carbohydrate source is pumped into a tank while correlating with monitoring the optical density and air is pumped into tank while correlating with the ethanol production, i.e. if ethanol is measured more oxygen is pumped into the tank.  
         [0148]     Inlets of air flow and carbohydrate are stopped after 16-18 hours. Cells are ready to be harvested after 24 hours.  
         [0149]     The cells are harvested and concentrated approximately 15 times by centrifugation (e.g. 4000 rpm, 10 minutes). 1% glycerol is added as cryoprotective agent.  
         [0000]     Spray/Fluid Bed Drying  
         [0150]     The concentrated cells is further up-concentrated in a fluid bed and thereafter pressed through an extruder or by spray drying directly into the dryer. The resulting yeast paste is then fed into a drying tower where the cells are dried. When the product is dry, it is distributed to packings of sterile aluminium bags in various sizes as follows.  
         [0151]     Spray/fluid bed dried products of  Saccharomyces cerevisiae  and  T. delbrueckii  are mixed in a desired ratio as described on pp, in package sizes of e.g. 500 g, 1 kg, 5 kg, 25 kg.  
         [0152]     Spray/fluid bed products of  Saccharomyces cerevisiae  and  K. thermotolerans  are mixed in a desired ratio as described on pp, in package sizes of e.g. 500 g, 1 kg, 5 kg, 25 kg.  
         [0153]     Spray/fluid bed products of  Saccharomyces cerevisiae, T. delbrueckii  and  K. thermotolerans  are mixed in a desired ratio as described on pp, in package sizes of e.g. 500 g, 1 kg, 5 kg, 25 kg.  
       Example 3  
       [0154]     Fermentation of Grape Juice with a Mixture of 2 Yeast Genera  
         [0155]     The following examples illustrate the preparation of an alcoholic fermentation by a mixed yeast starter culture, respectively.  
         [0000]     3.1. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae  and  T. delbrueckii  for Alcoholic Fermentation.  
         [0156]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0157]     A sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae  and  T. delbrueckii.    
         [0158]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0159]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae  and  T. delbrueckii. T. delbrueckii  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     3.2. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae  and  K.thermotolerans  for Alcoholic Fermentation.  
         [0160]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0161]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae  and  K. thermotolerans.    
         [0162]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0163]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae  and  K. thermotolerans. K. thermotolerans  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     3.3. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae  and  Candida  for Alcoholic Fermentation.  
         [0164]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0165]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae  and  Candida.    
         [0166]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0167]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae  and  Candida. Candida  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     3.4. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae  and  DekkeralBrettannomyces  for alcoholic fermentation.  
         [0168]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0169]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae  and  DekkeralBrettannomyces.    
         [0170]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0171]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae  and  DekkeralBrettannomyces. DekkeralBrettannomyces  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     3.5. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae  and  Pichia  for Alcoholic Fermentation.  
         [0172]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0173]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae  and  Pichia.    
         [0174]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0175]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae  and  Pichia. Pichia  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     3.6. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae  and  Metschnikowia  for Alcoholic Fermentation.  
         [0176]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0177]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae  and  Metschnikowia.    
         [0178]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0179]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae  and  Metschnikowia. Metschnikowia  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
       Example 4  
       [0000]     Fermentation of Grape Juice with a Mixture of 3 Yeast Genera  
         [0180]     The following examples illustrate the preparation of an alcoholic fermentation by a mixed yeast starter culture, respectively.  
         [0000]     4.1. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae, T. delbrueckii  and  K. thermotolerans  for Alcoholic Fermentation.  
         [0181]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0182]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae, T. delbrueckii  and  K. thermotolerans.    
         [0183]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0184]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae, T. delbrueckii  and  K. thermotolerans. T. delbrueckii  and  K. thermotolerans  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     4.2. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae, K. thermotolerans  and  Candida  for Alcoholic Fermentation.  
         [0185]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0186]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae, K. thermotolerans  and  Candida.    
         [0187]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0188]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae, K. thermotolerans  and  Candida. K. thermotolerans  and  Candida  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     4.3. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae, K. thermotolerans  and  DekkeralBrettannomyces  for Alcoholic Fermentation.  
         [0189]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0190]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae, K. thermotolerans  and  DekkeralBrettannomyces.    
         [0191]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0192]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae, K. thermotolerans  and  DekkeralBrettannomyces. K. thermotolerans  and  DekkeralBrettannomyces  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     4.4. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae, K. thermotolerans  and  Pichia  for Alcoholic Fermentation.  
         [0193]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0194]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae, K. thermotolerans  and  Pichia.    
         [0195]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0196]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae, K. thermotolerans  and  Pichia. K. thermotolerans  and  Pichia  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     4.5. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae, K. thermotolerans  and  Metschnikowia  for Alcoholic Fermentation.  
         [0197]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0198]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae, K. thermotolerans  and  Metschnikowia.    
         [0199]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0200]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae, K. thermotolerans  and  Metschnikowia. K. thermotolerans  and  Metschnikowia  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     4.6. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae, Candida  and  DekkeralBrettannomyces  for Alcoholic Fermentation.  
         [0201]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0202]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae, Candida  and  DekkeralBrettannomyces.    
         [0203]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0204]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae, Candida  and  DekkeralBrettannomyces. Candida  and  DekkeralBrettannomyces  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     4.7. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae, Candida  and  Pichia  for Alcoholic Fermentation.  
         [0205]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0206]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae, Candida  and  Pichia.    
         [0207]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0208]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae, Candida  and  Pichia. Candida  and  Pichia  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted. Saccharomyces cerevisiae will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     4.8. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae, Candida  and  Metschnikowla  for Alcoholic Fermentation.  
         [0209]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0210]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae, Candida  and  Metschnikowia.    
         [0211]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0212]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae, Candida  and  Metschnikowia. Candida  and  Metschnikowia  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted. Saccharomyces cerevisiae will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     4.9. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae, DekkeralBrettannomyces  and  Pichia  for Alcoholic Fermentation.  
         [0213]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0214]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae, DekkeralBrettannomyces  and  Pichia.    
         [0215]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0216]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae, DekkeralBrettannomyces  and  Pichia. DekkeralBrettannomyces  and Pichia will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     4.10. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae, DekkeralBrettannomyces  and  Metschnikowia  for Alcoholic Fermentation.  
         [0217]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0218]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae, DekkeralBrettannomyces  and  Metschnikowia.    
         [0219]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0220]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae, DekkeralBrettannomyces  and  Metschnikowia. DekkeralBrettannomyces  and  Metschnikowia  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     4.11. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae, T. delbrueckii  and  Candida  for Alcoholic Fermentation.  
         [0221]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0222]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae, T. delbrueckii  and  Candida.    
         [0223]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0224]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae, T. delbrueckii  and  Candida. T. delbrueckii  and  Candida  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     4.12. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae, T. delbrueckii  and  DekkeralBrettannomyces  for Alcoholic Fermentation.  
         [0225]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0226]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae, T. delbrueckii  and  DekkeralBrettannomyces.    
         [0227]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0228]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae, T. delbrueckii  and  DekkeralBrettannomyces. T. delbrueckii  and  DekkeralBrettannomyces  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     4.13. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae, T. delbrueckii  and  Pichia  for Alcoholic Fermentation.  
         [0229]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0230]     The sealed pouch is opened containing the dried mixed yeast starter culture of Saccharomyces cerevisiae, T delbrueckii and Pichia.  
         [0231]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0232]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae, T. delbrueckii  and  Pichia. T. delbrueckii  and  Pichia  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
         [0000]     4.14. Preparation of a Yeast Starter Culture of  Saccharomyces cerevisiae, T. delbrueckii  and  Metschnikowia  for Alcoholic Fermentation.  
         [0233]     The recommended dosage is corresponding to a final concentration of yeast cells of 1-2×10 6 /ml.  
         [0234]     The sealed pouch is opened containing the dried mixed yeast starter culture of  Saccharomyces cerevisiae, T. delbrueckii  and  Metschnikowia.    
         [0235]     A dosage of 10-20 g/hl grape juice to be fermented is transferred into a mixture of ⅓ of grape juice and ⅔ of tap water preheated to 35° C./95° F. at mixed. Leave without stirring for 20 minutes, then homogenize properly, and add into the grape juice during pumping.  
         [0000]     Fermentation  
         [0236]     The carbohydrates in the grape juice are then fermented into end-fermentation product by  Saccharomyces cerevisiae, T. delbrueckii  and  Metschnikowia. T. delbrueckii  and  Metschnikowia  will only be active in the first part of the alcoholic fermentation until the oxygen is depleted.  Saccharomyces cerevisiae  will be active through the alcoholic fermentation and will be responsible for the completion of the alcoholic fermentation.  
       REFERENCES  
       [0237]     Bauer, F. F. &amp; I. S. Pretorius. 2000. Yeast stress response and fermentation efficiency: How to survive the making of wine—A review.  South African Journal of Enology and Viticulture  21: 27-51.  
         [0238]     Hansen E. H., Nissen P., Sommer P., Nielsen J. C., Arneborg N. 2001. The effect of oxygen on the survival of non-Saccharomyces yeasts during mixed culture fermentations of grape juice with  Saccharomyces cerevisiae.  J. Applied Microbiology; 91: 541-547  
         [0239]     Kurtzman, C. P. and J. W. Fell (editors). 1998. The yeasts, a taxonomic study. 4th ed., Elsevier, N.Y.  
         [0240]     Pollien, P. et al., “Hyphenated headspace-gas chromatograhpy-sniffing technique: Screening or impact oderants and quantitative aromagram”,  Journal of Agriculture and Food Chemistryl,  45:2630-2637, 1997  
         [0241]     Williams, D. H. and Fleming, I.,  Mass Spectra I: Spectroscopic methods in organic chemistry,  5 th  ed. , p.170, Shoppenhangers Road, Berkshire, England