Abstract:
Described are novel mixtures of optical isomers of (i) styralyl alcohol (α-phenylethyl alcohol) or (ii) styralyl acetate (a-phenylethyl acetate), fermentation processes for preparing same and organoleptic uses for same as aroma or food flavor ingredients.

Description:
STATUS OF RELATED APPLICATIONS 
     This application is a divisional of U.S. Ser. No. 09/573,532, filed on May 19, 2000, now U.S. Pat. No. 6,511,686 B1, the contents hereby incorporated by reference as if set forth in its entirety. 
    
    
     BACKGROUND OF THE INVENTION 
     The present invention relates to novel styralyl alcohol isomer mixtures and novel sytralyl acetate isomer mixtures, fermentation processes for preparing same and organoleptic uses for same as aroma or food flavor ingredients. 
     In today&#39;s market, it is frequently desirable to identify flavor components of food items as being “natural flavors.” It is generally recognized in the industry that a flavor compound having been prepared by microbial processes can be designated as a natural product and therefore have an important place in the commercialization of products containing them. As a result, the industry has devoted considerable time and effort to develop methods for the production of flavoring components and, in particular, for the production of certain alcohol and acetate derivatives which can properly be called “natural.” 
     Furthermore, in today&#39;s market, a trend is developing whereby it is now determined to be desirable to identify fragrance ingredients of fragrance compositions as being “natural fragrance ingredients.” It is generally recognized in the industry that a fragrance compound having been prepared by microbial processes can be designated as a natural product and therefore have an important place in the commercialization of perfumes and perfumed articles containing them. As a result, the industry has devoted considerable time and effort to develop methods for the production of fragrance components and in particular for the production of alcohols and esters which can be called “natural fragrance ingredients.” 
     It is also well known in the flavor and fragrance industry that particular stereoisomers of specific ingredients are, in many instances, more advantageous than their corresponding opposite stereoisomers. Accordingly, a significant amount of research has been carried out covering the formation of such stereoisomers using, for example, microbial reduction. 
     Thus, Simon, et al,  Angew. Chem. Int. Ed. Engl . 24 (1985), pages 539-553 (title: “Chiral Compounds Synthesized by Biocatalytic Reductions”) discloses at section 3.2 on page 545 the hydrogenation of ketones to form chiral secondary alcohols using Clostridia such as  Clostridium kluyveri  and specifically sets forth the production of α-phenylethyl alcohol stereoisomers having the structure:                           
     with an enantiomeric excess of 94%. 
     Adlercreutz,  Biotechnology Letters , Volume 13, No. 4 at pages 229-234 (1991) (title: “ASYMMETRIC REDUCTION OF KETONES WITH ENZYMES FROM ACETIC ACID BACTERIA”) shows production of α-phenylethyl alcohol at page 233 using  G. oxydans  (an enantiomeric excess of 99%);  A. aceti  (an enantiomeric excess of 92%);  G. oxydans  (an enantiomeric excess of 75%);  A. aceti  (an enantiomeric excess of 86%);  A. pasteurianus  (an enantiomeric excess of 94%); and  A. peroxydans  (an enantiomeric excess of 66%) 
     Nakamura, et al,  Tetrahedron: Asymmetry , Volume 7, No. 10, at: pages 3021-3024, 1996, published by Elsevier Science Ltd. (title: “Asymmetric Synthesis of (S)-Arylalkanols by Microbial Reduction”) sets forth the production of the isomers having the structures:                           
     using acetophenone as a starting material in accordance with the reaction:                           
     using  Geotrichum candidum  IFO 4597, but only shows the production of a stereoisomer mixture having 28% enantiomeric excess. 
     Vicenzi, et al,  Enzyme and Microbial Technology , 20: pages 494-499, 1997, published by Elsevier Science Inc. (title: “Large-scale stereoselective enzymatic ketone reduction with in situ product removal via polymeric adsorbent resins”) discloses the stereoselective enzymatic reduction of 3,4-methylene-dioxyphenyl acetone to the corresponding S-3,4-methylene-dioxyphenyl isopropanol utilizing  Zygosaccharomyces rouxii.    
     Sorrilha, et. al,  Organic  &amp;  Medicinal Chemistry Letters , Volume 2, No. 2 at pages 191-196, 1992, published by Pergamon Press plc (title: “REDUCTION OF PHENYLKETONES BY IMMOBILIZED BAKER&#39;S YEAST”) discloses a process wherein baker&#39;s yeast immobilized on chrysotile and montmorillonite causes stereoselective reduction of 1-phenyl-1,2-propanedione to the corresponding (1R,2S)-diol. 
     Nothing in the prior art, however, discloses the production of the stereoisomeric mixture of the compounds having the structures:                           
     having an          α   D   20     =       -   38.6        °                            
     with an enantiomeric excess percent of 87.5 ([εε%=87.5]) and furthermore, nothing in the prior art discloses the production of the stereoisomeric mixture of esters having the structures:                           
     having        (       α   D   20     =       +   73.06        °       )                          
     with an enantiomeric excess percent of 79.2, [εε%=79.21] or          α   D   20     =       -   102.7        °                            
     with an enantiomeric excess percent of 89.15, [εε%=89.15], which materials have been found by us to have unexpected, unobvious and advantageous properties insofar as their organoleptic (flavor and fragrance) utilities are concerned. 
     SUMMARY OF THE INVENTION 
     The above and other objects and features of the invention are obtained in accordance with the present invention by carrying out a process using (i) reductive reaction techniques to produce and recover certain naturally occurring alcohols and (ii) esterification reaction techniques to produce and recover certain naturally occurring esters, found to be useful for their organoleptic properties in augmenting or enhancing the aroma or taste of consumable materials such as foodstuffs, chewing, gums, toothpastes, oral care products, chewing tobaccos, smoking tobaccos, perfume compositions, colognes and perfumed articles such as solid or liquid detergents, perfumed polymers, fabric softener compositions, fabric softener articles, cosmetic powders, hair preparations and the like. 
     The reductive reaction products are styralyl alcohol stereoisomer mixtures containing compounds having the structures:                           
     produced according to a fermentation reaction whereby acetophenone is reduced according to the reaction:                           
     using a culture of  Kluyveromyces polysporus  ATCC 22028. 
     The resulting products may be used as is for their organoleptic properties or may be further reacted by means of esterification with acetic acid according to the reaction:                           
     The aforementioned esterification reaction may take place using such esterification catalysts as citric acid or may take, place by means of a fermentation reaction using an ester-forming enzyme, preferably  Candida antarctica  esterase expressed in  Asperigilus orzae , for example NOVOZYM® 435, a triacylglycerol hydrolase (E.C. No.3.1.1.3) acting as an effective carboxylesterase. NOVOZYM® is a trademark of the Novo Nordisk A/S Organization of Novo Alle, 2880Bagsvaerd, Denmark. 
     When carrying out the reductive fermentation reaction, to wit:                           
     the resulting product has an optical rotation of −38.6°, [α=−38.6°] and an enantiomeric excess percent of about 87.5, [εε%=87.5±0.5]. 
     When carrying out the esterification reaction, to wit:                           
     using a citric acid catalyst, the resultant stereoisomer mixture has an optical activity of −102.7°, [α=−102.7°] with an enantiomeric excess percent of about 78.31, [εε%=78.31±0.5]. 
     When carrying out the estenfication reaction, to wit:                           
     using the enzyme  Candida antarctica  esterase expressed in Asperigilus orzae, specifically NOVOZYM® 435, a triacylglycerol hydrolase (E.C. No. 3.1.1.3) acting as an effective carboxylesterase, a mixture of stereoisomers having an optical activity of +73.06° is produced ([α=+73.06°]) with an enantiomeric excess percent of about 81.1, [εε%=81.1±0.5]. 
     The mixture of optical isomers of the styralyl alcohol having the structures:                           
     produced as set forth above having an optical rotation of −38.6°, [α=−38.6°] with an enantiomeric excess percent of about 87.5, [εε%=87.5±0.5] has a mild hyacinth, gardenia aroma with strawberry nuances. It is, accordingly, useful in augmenting, enhancing and imparting aroma, in or to floral fragrances and is useful in the creation of strawberry-flavored foodstuffs; e.g., strawberry-flavored gelatin desserts, strawberry-flavored chewing, gums and strawberry-flavored fruit preparations for yogurt. 
     The mixture of stereoisomers of esters having the structures:                           
     prepared as set forth above using a citric acid catalyst and having an optical rotation of −102.7°, [α=102.7°] and an enantiomeric excess percent of about 78.31, [εε%=78.31±0.5] has a fresh strawberry, green, dried fruit aroma with a green, avocado taste profile and strawberry jam nuances. From a fragrance standpoint, this substance is described as having a strawberry, green, dried fruit aroma with strawberry topnotes. 
     The mixture of stereoisomeric esters having the structures:                           
     prepared as set forth above using the ester-forming enzyme,  Candida antarctica  esterase expressed in  Asperigilus orzae , and having an optical rotation of +73.06°, [α=+73.06°] with an enantiomeric excess percent of about 81.1, [εε%=81.1±0.5] has a fruity, floral, jasmine, mimosa, gardenia aroma with apricot, apple and strawberry jam flavor nuances. This material is also useful in the formation of fruity, floral, jasmine fragrances, as well as strawberry-flavored foodstuffs and food preparations such as strawberry-flavored gelatin desserts, strawberry-flavored ice cream and strawberry-flavored preparations for yogurts. 
     In carrying out the process for production of the styralyl alcohol stereoisomer mixtures containing stereoisomers having the structures:                           
     an inoculum preparation is first prepared containing a culture of  Kluyveromyces polysporus  ATCC 22028 and also containing nutrients including dextrose. The inoculum, after incubation, is then placed in a production fermenter which is aerated and agitated at, for example, an aeration rate of 0.25 v/v/m; at a temperature of, for example, 25° C.; and at an agitation rate of, for example, 300 rpm for a time period of, for example, 24 hours. As set forth in the Detailed Description of the invention, infra, the pH range may vary from about 5.5 up to about 6. 
     The resulting product is then extracted with a solvent such as ethyl acetate, and the solvent extract is then washed in order to bring the pH up to approximately neutral (pH=7). The resulting washed extract is then concentrated and the resulting concentrate is then fractionally distilled. The resulting distillate is then used “as is” for its organoleptic properties or further reacted with acetic acid according to the reaction:                           
     When carrying out the reaction using a citric acid catalyst, the resulting distillate is admixed with acetic acid and citric acid, and the resulting mixture is refluxed for about 15 hours while excess acetic acid is recovered. At the end of the 15 hour period, the resulting product is fractionally distilled. 
     When carrying out the esterification reaction, to wit:                           
     using the: ester-forming; enzyme,  Candida antarctica  esterase expressed in  Asperigilus orzae , specifically, NOVOZYM® 435, the mixture of styralyl alcohol stereoisomers having the structures:                           
     is admixed with a small amount of water and a small amount of NOVOZYM® 435. The resulting mixture is heated to 40° C. and acetic acid is then added over a period of between about 5 and about 10 hours and then continuously stirred for an additional 40-60 hours. The conversion using the “enzyme” process is between about 2.5 and about 3.5%. The resulting product is then fractionally distilled at a pressure of 5 mm/Hg and at a head temperature of 34-38° C. 
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS 
     The present invention will be further understood with reference to the accompanying drawings wherein: 
     FIG. 1A is a GC-mass spectrum for the reaction product for Example I containing the mixture of stereoisomers having the structures:                           
     FIG. 1B is another GC-mass spectrum for the mixture of stereoisomers having the structures:                           
     produced according to Example I. 
     FIG. 1C is a GLC profile for the reaction product of Example I containing a mixture of stereoisomers having the structures:                           
     FIG. 1D is a mass spectrum for the product of Example I containing a mixture of stereoisomers having the structures:                           
     FIG. 1E is a GLC profile for the reaction product of Example I containing the stereoisomers having the structures:                           
     FIG. 1F is a mass spectrum for the reaction product of Example I containing the stereoisomers having the structures:                           
     FIG. 2A is a GLC profile for the reaction product of Example II containing the stereoisomers having the structures:                           
     FIG. 2B is a “history plot” for the product formed according to the process of Example II which contains and consists essentially of the stereoisomers having the structures:                           
     FIG. 2C is a GC-mass spectrum for the reaction product of Example II containing the stereoisomers having the structures:                           
     FIG. 2D is a GC-mass spectrum for the reaction product of Example II containing the stereoisomers having the structures:                           
     FIG. 3A is a chiral chromatogram for the reaction product of Example IV containing the stereoisomers having the structures:                           
     with the optical rotation of +73.06°, [α=+73.06°] and with an enantiomeric excess percent of about 81.1, [εε%=81.1±0.5]. 
     FIG. 3B is another chiral GC chromatogram for the reaction product of Example IV containing a mixture of stereoisomers having the structures:                           
     with an optical rotation of +73 06°, [α=+73.06°] and having an enantiomeric excess percent of about 81.1, [εε%=81.1±0.5]. 
     FIG. 3C is a GLC profile for the reaction product of Example IV containing stereoisomers having the structures:                           
     FIG. 4 represents a cutaway side elevation view of apparatus used in forming perfumed polymers which contain embedded therein at least one of the stereoisomers having one of the structures:                           
     FIG. 5 is a front view of the apparatus of FIG. 4 looking in the direction of the arrows along lines 5—5. 
     FIG. 6A is a GC-mass spectrum (using a Carbowax column) for the reaction product of Example III containing a mixture of stereoisomers having the structures:                           
     with the mixture having an optical rotation of −102.7°, [α=−102.7°] and having an enantiomeric excess percent of about 78.31, [εε%=78.31±0.5]. 
     FIG. 6B is a GC-mass spectrum for the reaction product of Example III (using an OV- 1  column) containing the stereoisomers having the structures:                           
     with the mixture having an optical rotation of −102.7°, [α=−102.7°] and an enantiomeric excess percent of about 78.31, [εε%=78.31±0.5]. 
    
    
     DETAILED DESCRIPTION OF THE DRAWINGS 
     Referring to FIG. 1D, the peak indicated by reference numeral  10  is the peak for acetic acid. The peak indicated by reference numeral  11  is the peak for ethyl acetate. The peak indicated by reference numeral  12  is for isobutyl alcohol. The peak indicated by reference numeral  13  is for 3-methyl-butanol. The peak indicated by reference numeral  14  is for a mixture of acetophenone and the mixture of stereoisomers having the structures:                           
     The peak indicated by reference numeral  15  is for phenylethyl alcohol having the structure:                           
     The peak indicated by reference numeral  16  is for the mixture of stereoisomers having the structures:                           
     The peak indicated by reference numeral  17  is the peak for acetophenone. The peak indicated by reference numeral  18  is for the compound having the structure:                           
     The peak indicated by reference numeral  19  is the peak for β-phenylethyl acetate having the structure:                           
     Referring to FIG. 1F, the peak indicated by reference numeral  20  is the peak for ethyl acetate. The peak indicated by reference numeral  21  is for isobutyl alcohol. The peak indicated by reference numeral  22  is for 3-methyl-butanol. The peak indicated by reference numeral  23  is for acetic acid. The peak indicated by reference numeral  24  is for 2-methyl-propionic acid. The peak indicated by reference numeral  25  is for acetophenone. The peak indicated by reference numeral  26  is for the mixture of stereoisomers having the structures:                           
     The peak indicated by reference numeral  27  is the peak for β-phenylethyl alcohol having the structure:                           
     Referring to FIG. 3A, the peaks indicated by reference numerals  31  and  32  are for the stereoisomers having the structures:                           
     The peak indicated by reference numeral  30  is for the mixture of stereoisomers having the structures:                           
     With reference to FIG. 3B, the peaks indicated by reference numerals  34  and  35  are for the stereoisomers having the structures:                           
     separately. The peak indicated by reference numeral  33  is for the mixture of stereoisomers having the structures:                           
     With reference to FIG. 3C, the peak indicated by reference numeral  36  is the peak for the mixture of stereoisomers having the structures:                           
     The peak indicated by reference numeral  37  is the peak for the mixture of stereoisomers having the structures:                           
     Referring to FIGS. 4 and 5, there is provided a process for forming scented polymer elements (wherein the polymer may be a thermoplastic polymer such as low density polyethylene or polypropylene or copolymers of ethylene and vinyl acetate or mixtures of polymers and copolymers such as copolymers of ethylene and vinyl acetate and polyethylene) such as pellets useful in the formation of plastic particles useful in fabricating certain articles which may be perfumed. This process comprises heating the polymer or mixture of polymers to the melting point of said polymer or mixture of polymers, e.g., 250° C. in the case of low density polyethylene. The lower most portion of the container is maintained at a slightly lower temperature, and the material in the container is taken off at such location for delivery through the conduit (indicated by reference numeral  218 ). Thus, referring to FIGS. 4 and 5 in particular, the apparatus used in producing such elements comprises a device for forming the polymer containing perfume, e.g., polyethylene or polyethylene-polyvinyl acetate or mixtures of same or polypropylene, which comprises a vat or container  212  into which the polymer taken alone or in admixture with other copolymers and the perfuming substance, which is at least one of the stereoisomers having one of the structures:                           
     of our invention and other compatible perfumes, is placed. The container is closed by means of an airtight lid  228  and clamped to the container by bolts  265 . A stirrer  273  traverses the lid or cover  228  in an airtight manner and is rotatable in a suitable manner. A surrounding cylinder  212 A having heating coils which are supplied with electric current through cable  214  from a rheostat or control  216  is operated to maintain the temperature inside the container  212  such that the polymer in the container will be maintained in the molten or liquid state. It has been found advantageous to employ polymers at such a temperature that the viscosity will be in the range of 90-100 Saybolt seconds. The beater is operated to maintain the upper portion of the container  212  within a temperature range of, for example, 220°-270° C. in the case of low density polyethylene. The bottom portion of the container  212  is heated by means of heating coils  212 A regulated through the control  220  connected thereto through a connecting wire  222  to maintain the lower portion of the container  212  within a temperature range; of 220°-270° C. 
     Thus, the polymer or mixture of polymers added to the container  212  is heated from 10-12 hours, whereafter the perfume composition or perfume material which, contains one or more of the stereoisomers having the structures:                           
     of our invention is quickly added to the melt. Generally, about 10-45% by weight of the resulting mixture of the perfumery substance is added to the polymer. 
     After the perfume material is added to the container  212 , the mixture is stirred for a few minutes, for example, 5-15 minutes, and maintained within the temperature ranges indicated previously by the heating coils  212 A. The controls  216  and  220  are connected through cables  224  and  226  to a suitable supply of electric current for supplying the power for heating purposes. 
     Thereafter, the valve “V” is opened permitting the mass to flow, outwardly through conduit  232  (also shown by reference numeral  218  (in cutaway cross section)) having a multiplicity of orifices  234  adjacent to the lower side thereof. The outer end of the conduit  232  is closed so that the liquid polymer in intimate admixture with one or more of the stereoisomers having one of the structures:                           
     and one or more other substances (if desired) will continuously drop through orifices  234  downwardly from the conduit  232 . During this time, the temperature of the polymer intimately admixed with the perfumery substance in the container  212  is accurately controlled so that a temperature in the range of form about 240°-250° C., for example (in the case of low density polyethylene) will exist in the conduit  232 . The regulation of the temperature through the controls  216  and  220  is essential in order to insure temperature balance to provide for the continuous dripping or dropping of molten polymer intimately admixed with the perfume substance which contains one or more of the stereoisomers having one of the structures:                           
     of our invention, through the orifices  234  at a rate which will insure the formation of droplets  236  which will fall downwardly onto a moving conveyor belt  238  caused to run between conveyor wheels  240  and  242  beneath the conduit  232 . 
     When the droplets  236  fall onto the conveyor  238  they form pellets  244  which harden almost instantaneously and fall off the end of the conveyor  238  into a container  245  which is advantageously filled with water  252  or some other suitable cooling liquid to insure the rapid cooling of each of the pellets  244 . The pellets  244  are then collected from the container  245  and utilized for the formation of other functional products, e.g., garbage bags and the like. 
     A feature of the invention is the provision for the moistening of the conveyor belt  238  to insure the rapid formation of the solid polymer-aromatizing agent containing pellets  244  without sticking to the belt. The belt  238  is advantageously of a material which will not normally stick to a melted polymer, but the moistening means  248  insures a sufficiently cold temperature of the belt surface for the adequate formation of the pellets  244 . The moistening means comprises a container  250  which is continuously fed with water  252  to maintain a level  254  for moistening a sponge element  256  which bears against the exterior surface of the belt  238 . 
     DETAILED DESCRIPTION OF THE INVENTION 
     The reactions according to the present invention are shown thusly:                           
     More specifically, the first reaction involving reduction of a ketone group to form a secondary alcohol is carried out in the presence of  Kluyveromyces polysporus  ATCC 22028. 
     The process is carried out by first introducing an inoculum containing the said  Kluyveromyces polysporus  ATCC 22028 into a reaction vessel which contains a production medium, typically including a nutrient source, a buffering agent such as KH 2 PO 4 , a yeast extract and other nutrient sources which may include trace minerals and growth factors. 
     Immediately, subsequent to the introduction of the inoculum, a source of dextrose in a suitable nutrient medium is begun feeding into the reaction vessel (and throughout the entire fermentation procedure). 
     Up to about 30 hours after inoculation, acetophenone having the structure:                           
     is pumped into the reaction vessel. Incubation is then continued at a temperature in the range of 20° C. up to 30° C., at an agitation rate: of from about 250 up to about 350 rpm and an aeration rate of from about 0.20 up to about 0.40 v/v/m. Conversion to the resulting mixture of stereoisomers having the structures.                           
     is monitored by means of GC. 
     At the conclusion of the fermentation, the fermentation broth is pasteurized for a period of between about 30 up to about 70 minutes at a temperature of between about 60° C. and about 75° C., cooled and stored. The crude product is recovered by extraction of the whole broth using an equal volume of a suitable solvent preferably ethyl, acetate. The spent broth is extracted a second time and the recovered ethyl acetate layers are combined and washed with aqueous sodium chloride, preferably. The solvent is then evaporated using such equipment as a rotary evaporator at a water bath temperature in the range of from about 35° C. up to about 50° C. under vacuum. The crude extract is then further purified, if desired, for use for its organoleptic properties; or, in the alternative, the crude extract may be converted to styralyl acetate directly without any further purification; or, optionally, the resulting distillate can be converted to styralyl acetate. 
     Using the aforementioned procedure, the optical rotation of the resulting mixture of stereoisomers having the structures:                           
     is −38.6° with an enantiomeric excess percent of about 87.5, [εε%=87.5±0.5]. The aforementioned term is intended to mean that the enantiomeric excess percent is between 87 and 88%. 
     If desired, the resulting mixture of stereoisomers having the structures:                           
     may further be reacted with acetic acid in order to form the resulting esters having the structures:                           
     As stated, supra, and as exemplified, infra, the aforementioned esters may be formed, in the alternative, into a mixture of stereoisomers having an optical rotation of 102.7°, [α=−102.7°] with an enantiomeric excess percent of about 78.31, [εε%=78.31±0.5] when carrying out the esterification with a citric acid catalyst or the mixture of esters may have an optical rotation of +73.06° with an enantiomeric excess percent of about 81.1, [εε%=81.1±0.5] when using an ester-forming enzyme which is  Candida antarctica  esterase expressed in  Asperigilus orzae , preferably NOVOZYM® 435, a triacylglycerol hydrolase (E.C. No. 3.1.1.3) acting as an effective carboxylesterase. 
     When carrying out the reaction:                           
     using a citric acid catalyst, excess acetic acid is admixed with the stereoisomers having the structures:                           
     and a small amount of citric acid, e.g., approximately 0.1% of the weight of the entire reaction mass. The resulting mixture is then refluxed for a period of time of between about 10 up to about 30 hours. During such refluxing, a substantial quantity of the excess acetic acid is recovered by means of azeotropic distillation with water of reaction. The reaction is carried out at a reflux temperature of approximately 115-120° C. at atmospheric pressure. 
     At the end of the reaction, the reaction product is fractionally distilled to yield a mixture of isomers having the structures:                           
     wherein the mixture has an optical rotation of −102.7°, [α=102.7°] with an enantiomeric excess percent: of about 78.31, [εε%=78.31±0.5]. 
     In the alternative, in place of the citric acid catalyst, a fermentation esterification can be carried out using the ester-forming enzyme,  Candida antarctica  esterase expressed in  Aspergilus orzae , preferably NOVOZYM® 435; a triacylglycerol hydrolase (E.C. No. 3.1.1.3) acting as an effective carboxylesterase. 
     In carrying out the reaction using the ester-forming enzyme, the reaction may be carried out with, the ester-forming enzyme, the acetic acid and the styralyl alcohol with a small amount of water, or the reaction may be carried out in the presence of an inert solvent such as hexane. We have found that when using the inert solvent such as hexane, a higher conversion is achieved. Thus, initially, styralyl alcohol, water and the ester-forming catalyst are initially admixed and the resulting mixture is heated to a temperature of between about 35° C. and 45° C. The resulting mixture is then further admixed with acetic acid which is added over a period of between about 5 and 10 hours while maintaining the reaction mass at 40° C. Optionally, solvents such as hexane may be introduced into the mixture of styralyl alcohol and water prior to the introduction of acetic acid. When a solvent such as hexane is utilized, the conversion of the styralyl alcohol to the ester is between about 7 and about 10%. However, when such solvent is not used, the conversion is between about 2.5 up to about 3.5%. 
     Accordingly, it is preferred that a solvent such as n-hexane be utilized in carrying out the esterification reaction using an ester-forming enzyme. The amount of such solvent as n-hexane is approximately equal to the weight of styralyl alcohol in the reaction mass. 
     The mixtures of stereoisomers of styralyl alcohol and styralyl acetate obtained in accordance with the present invention and one or more auxiliary perfume ingredients including, for example, hydrocarbons, alcohols other than the styralyl alcohols of our invention, ketones, aldehydes, nitriles, esters other than the styralyl acetates of our invention, ethers, synthetic essential oils, lactones and natural essential oils may be admixed so that the combined odors of the individual components produce a pleasant and desired fragrance, particularly and preferably in the fruity and floral area (e.g., jasmine and gardenia aromas). Such perfume compositions usually contain (a) the main note or the “bouquet” or foundation stone of the composition (b) modifiers which round off and accompany the main note; (c) fixatives which include odorous substances which lend a particular note to the perfume throughout all stages of evaporation and substance which retard evaporation; and (d) topnotes, which are usually low boiling, fresh-smelling materials. 
     In perfume compositions, it is the individual compositions which contribute to their particular olfactory characteristics; however, the overall sensory effect of the perfume compositions will be at least the sum total of the effects of each of the ingredients. Thus, one or more of the styralyl acetate or styralyl alcohol stereoisomer mixtures of our invention can be used to alter, modify or enhance the aroma characteristics of a perfume composition, for example, by utilizing or moderating the olfactory reaction contributed by another ingredient in the composition. 
     The amount of styralyl alcohol or styralyl acetate stereoisomer composition of our invention, which will be effective in perfume compositions as well as in perfumed articles and colognes, depends upon many factors including the other ingredients, their amounts and the effects which are desired. It has been found that perfume compositions containing as little as 0.005% of the styralyl alcohol or styralyl acetate stereoisomers or even less (e.g., 0.0025%) can be used to impart green, floral, hyacinth, gardenia, jasmine, mimosa, fruity, dried fruit and strawberry aromas with green, fruity, strawberry, “fresh air-dried clothing” topnotes to soaps, cosmetics, detergents including anionic, cationic, nonionic and zwitterionic solid or liquid detergents, perfumed polymers and other products. The amount employed can range up to 70% of the fragrance and will depend upon consideration of cost, nature of the end product, the effect desired on the finished product and the particular fragrance sought. 
     The mixtures of styralyl alcohol stereoisomers and styralyl acetate stereoisomers of this invention are useful when taken alone or taken together with other perfumery ingredients in detergents, soaps, space ordorants and deodorants, perfumes, colognes, toilet waters, bath preparations, hair preparations such as lacquers, brilliantines, pomades, shampoos; cosmetic preparations such as creams, deodorants, hand lotions and sun screens; powder such as talcs, dusting powders, face powders and the like. 
     As little as 0.25% of the mixtures of styralyl alcohol stereoisomers or styralyl acetate stereoisomers can suffice to impart intense and substantive green, floral, hyacinth, gardenia, jasmine, mimosa, fruity, dried fruit and strawberry aromas with green, fruity, strawberry and “fresh air-dried clothing” topnotes to floral and fruity perfume formulations. Generally, no more than 5% of the mixtures of styralyl alcohol stereoisomers or styralyl acetate stereoisomers based on the ultimate end product is required to be used in the perfume compositions. 
     Furthermore, as little as 0.25% of the mixture of styralyl alcohol stereoisomers or styralyl acetate stereoisomers can suffice to impart such aromas to perfumed articles, per se, whether in the presence of other perfume materials or whether used by themselves. Thus, the range of use of the mixtures of styralyl alcohol stereoisomers or styralyl acetate stereoisomers of this invention in perfumed articles, e.g., perfumed polymers and solid or liquid anionic, cationic, nonionic or zwitterionic solid or liquid detergents may vary from 0.25% up to about 5% by weight based on the total weight of the perfumed article. 
     In addition, the perfume composition or fragrance composition of our invention can contain a vehicle or carrier for the mixture of styralyl alcohol stereoisomers or styralyl acetate stereoisomers of our invention. The vehicle can be a liquid such as a nontoxic alcohol (e.g., ethanol); a nontoxic glycol (e.g., propylene glycol) or the like. The carrier can also be an absorbent solid such as a gum (e.g., gum arabic, xanthan gum or guar gum) or components for encapsulating the composition by means of coacervation (such as by gelatin) or by means of the formation of a polymer around a liquid center. This can be accomplished by using a urea formaldehyde prepolymer to form a polymeric capsule around a liquid perfume composition center as is known in the prior art. 
     It will be appreciated from the present disclosure that the mixtures of styralyl alcohol stereoisomers or styralyl acetate stereoisomers according to the present invention can be used to alter, vary, fortify, modify, enhance or otherwise improve the flavor of a wide variety of materials which are ingested, consumed or otherwise organoleptically sensed. 
     The terms “alter” and “modify” in their various forms will be understood herein to mean the supplying or imparting of a flavor character or note to an otherwise bland, relatively tasteless substance or augmenting an existing flavor characteristic where the natural flavor is deficient in some regard or supplement the existing flavor impression to modify its organoleptic character. 
     The term “enhance” is intended herein to mean the intensification (by use, of the mixtures of styralyl alcohol stereoisomers or styralyl acetate stereoisomers of this invention) of a flavor, aroma note or nuance in a foodstuff, perfume composition or perfumed article without changing the quality of said note or nuance. 
     A “flavoring composition” as referred to herein means one which contributes a part of the overall flavor impression by supplementing or fortifying a natural or artificial flavor in a material or one which supplies substantially all the flavor and/or aroma character to a consumable article. 
     The term “foodstuff” as used herein includes both solid and liquid ingestible material for man or animals, which materials usually do, but need not have nutritional value. Thus, foodstuffs include meats; gravies; soups; convenience foods; malt; alcoholic and other beverages; milk and dairy products; seafood including fish, crustaceans, mollusks and the like; candies; vegetables; cereals; soft drinks; snacks; dog and cat foods; other veterinary products and the like. 
     When the mixtures of styralyl alcohol stereoisomers or styralyl acetate stereoisomers of this invention are used in a flavoring composition, they can be combined with conventional flavoring materials or adjuvants. Such co-ingredients or flavoring adjuvants are well known in the art for such and have been extensively described in the literature. Requirements of such adjuvant materials are: 
     (1) that they be nonreactive with the styralyl alcohol stereoisomer mixtures or styralyl acetate stereoisomer mixtures of this invention; 
     (2) that they be organoleptically compatible with the mixtures of styralyl alcohol stereoisomers or styralyl acetate stereoisomers of this invention whereby the flavor of the ultimate consumable material to which the styralyl alcohol stereoisomers or styralyl acetate stereoisomers are added is not detrimentally affected by the use of the adjuvant; and 
     (3) that they be ingestible, acceptable and thus nontoxic or otherwise nondeleterious. 
     Apart from these requirements, conventional materials can be used and broadly include other flavor materials, vehicles, stabilizers, thickeners, surface active agents, conditioners and flavor intensifiers. 
     Such conventional flavoring materials include saturated fatty acids; unsaturated fatty acids and amino acids; alcohols (other than the styralyl alcohol stereoisomers of our invention) including primary and secondary alcohols; esters (other than the styralyl acetate stereoisomer mixtures of our invention); carbonyl compounds including ketones and aldehydes; lactones; other cyclic organic materials including benzene derivatives, alicyclic compounds, heterocyclics such as furans, pyridines, pyrazines and the like; sulfur-containing compounds including thiols, sulfides, disulfides and the like; proteins; lipids; carbohydrates; so-called flavor potentiators such as monosodium glutamate, magnesium glutamate, calcium glutamate, guanylates, and inosinates; natural flavoring materials such as cocoa, vanilla and caramel; essential oils and extracts such as anise oil, clove oil and the like; and artificial flavoring materials such as vanillin and the like. 
     Specific preferred flavor adjuvants are as follows: 
     anise oil; 
     ethyl-2-methyl butyrate; 
     vanillin; 
     cis-3-heptenol; 
     cis-3-hexenol; 
     trans-2-heptenol; 
     cis-3-heptenal; 
     butyl valerate; 
     2,3-diethyl pyrazine; 
     methyl cyclopentenolone; 
     benzaldehyde; 
     valerian oil; 
     3,4-dimetboxyphenol; 
     amyl acetate; 
     amyl cinnamate; 
     γ-butyryl lactone; 
     furfural; 
     trimethyl pyrazine; 
     phenyl acetic acid; 
     isovaleraldehyde; 
     ethyl maltol; 
     ethyl vanillin; 
     ethyl valerate; 
     cocoa extract; 
     coffee extract; 
     peppermint oil; 
     spearmint oil; 
     clove oil; 
     anethol; 
     cardamom oil; 
     wintergreen oil; 
     cinnamic aldehyde; 
     ethyl-2-methyl valerate; 
     γ-hexenyl lactone; 
     2,4-decadienal; 
     2,4-heptadienal; 
     2-methyl-2-pentenoic acid; 
     2-methyl-3-pentenoic acid; 
     the ethyl ester of 2-methyl-2-pentenoic acid; 
     the ethyl ester of 2-methyl-3-pentenoic acid; 
     the ethyl ester of 2-methyl4-pentenoic acid; 
     2-methyl4-pentenoic acid; 
     the hexyl ester of 2-methyl-3-pentenoic; and 
     butylidene phthalide. 
     The following table sets forth organoleptic properties of the mixtures of styralyl alcohol stereoisomers and styralyl acetate stereoisomers of our invention: 
     
       
         
               
               
               
             
           
               
                 TABLE I 
               
               
                   
               
               
                   
                 Fruit 
                   
               
               
                 Description of Composition 
                 Flavor Properties 
                 Fragrance Properties 
               
               
                   
               
             
             
               
                 Mixture of stereoisomers 
                 A strawberry flavor. 
                 A hyacinth, gardenia 
               
               
                 having the structures: 
                   
                 aroma with strawberry 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
                   
                 topnotes. 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                   
               
               
                 having an optical rotation 
               
               
                 of −38.6°, [α = −38.6°] 
               
               
                 and an enantiomeric excess 
               
               
                 percent of about 
               
               
                 87.5, [εε% = 87.5 ± 0.5]. 
               
               
                 Mixture of stereoisomers 
                 A strawberry jam, 
                 A fruity, floral, 
               
               
                 having the structures: 
                 apricot and 
                 jasmine, mimosa and 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 apple flavor. 
                 gardenia aroma with strawberry, fruity and green topnotes. 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                   
               
               
                 having an optical rotation 
               
               
                 of +73.06°, [α = +73.06°] 
               
               
                 and an enantiomeric excess 
               
               
                 percent of about 
               
               
                 81.1, [εε% = 81.1 ± 0.5]. 
               
               
                 Mixture of stereoisomers 
                 A strawberry jam, 
                 A strawberry, green, 
               
               
                 having the structures: 
                 green, dried fruit, 
                 dried fruit aroma with 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 avocado taste. 
                 strawberry, green, “fresh air-dried clothing” topnotes. 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                   
               
               
                 having an optical rotation 
               
               
                 of −102.7°, [α = −102.7°] 
               
               
                 and an enantiomeric excess 
               
               
                 percent of about 
               
               
                 78.31, [εε% = 78.31 ± 0.5]. 
               
               
                   
               
             
          
         
       
     
     The following Examples I, II, III and IV illustrate methods of our invention used to produce the mixtures of stereoisomers of styralyl alcohol and mixtures of stereoisomers of styralyl acetate of our invention. Examples V, et seq, serve to illustrate the organoleptic utilities of the mixtures of stereoisomers of styralyl alcohol and mixtures of stereoisomers of styralyl acetate of our invention. All parts and percentages given herein are by weight unless otherwise specified. 
     EXAMPLE I 
     PRODUCTION OF STYRALYL ALCOHOL IN LABORATORY FERMENTERS 
     I. Inoculum Preparation 
     A. Medium: 
     The following medium was prepared by admixing the following ingredients: 
     
       
         
               
               
             
               
             
           
               
                   
               
               
                 g/L 
                 Ingredients 
               
               
                   
               
             
             
               
                 5.0 
                 AMBERFERM ® 4000 Peptone 
               
               
                 5.0 
                 TASTONE ® 900 
               
               
                 5.0 
                 KH 2 PO 4  (potassium phosphate, monobasic) 
               
               
                 5.0 
                 Malt extract 102 
               
               
                 0.5 
                 MgSO 4 ·7H 2 O (magnesium sulfate, heptahydrate) 
               
               
                 1 L 
                 Deionized water 
               
               
                 50.0  
                 CERELOSE ® 2001 (added after sterilization) (dextrose) 
               
             
          
           
               
                 pH = 7.0, adjusted before sterilization with 25% aqueous NaOH solution 
               
               
                   
               
             
          
         
       
     
     B. Parameters: 
     
       
         
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 Temperature: 
                 25° C.; 
               
               
                   
                 Agitation: 
                 100 rpm; and 
               
               
                   
                 Duration: 
                 24 hours. 
               
               
                   
                   
               
             
          
         
       
     
     A 2.8 liter Fernbach flask containing 500 ml of the inoculum medium of A, supra, was sterilized at 121° C. for 30 minutes. The flask was inoculated with 1.8 ml of a frozen culture of  Kluyveromyces polysporus  ATCC 22028; 50.0 ml of sterile 50% dextrose (CERELOSE® 2001) and 2-3 drops of antifoam were added. The flask was inoculated in a shaker (100 rpm) at 25° C. for 24 hours. 
     II. Production 
     A. Medium: 
     The following medium was prepared by admixing the following ingredients: 
     
       
         
               
               
             
               
             
           
               
                   
               
               
                 g/L 
                 Ingredients 
               
               
                   
               
             
             
               
                 50.0 
                 AMBERFERM ® 4000 Peptone 
               
               
                 50.0 
                 TASTONE ® 900 
               
               
                 50.0 
                 Malt extract 102 
               
               
                 50.0 
                 KH 2 PO 4  (potassium phosphate, monobasic) 
               
               
                  5.0 
                 MgSO 4 ·7H 2 O (magnesium sulfate, heptahydrate) 
               
               
                 10.0 
                 Antifoam Hi MARS-010 ® FGK 
               
               
                      11.0 L 
                 Deionized water 
               
               
                 500.0  
                 CERELOSE ® 2001 (added after sterilization) (dextrose) 
               
             
          
           
               
                 pH = 7.0, adjusted before sterilization with 25% aqueous NaOH solution 
               
               
                   
               
             
          
         
       
     
     B. Fermenter Parameters: 
     
       
         
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 Inoculum: 
                 500 ml of propagation culture; 
               
               
                   
                 Broth Volume: 
                 11.0 liters; 
               
               
                   
                 Temperature: 
                 25° C.; 
               
               
                   
                 Aeration: 
                 0.25 v/v/m; 
               
               
                   
                 Agitation: 
                 300 rpm (impeller diameter 84 mm); and 
               
               
                   
                 Duration: 
                 24 hours. 
               
               
                   
                   
               
             
          
         
       
     
     C. Process: 
     A total of 10 liters of medium (with an additional 1 liter of water added to make up for steam lost during sterilization) was sterilized in a 23 liter fermenter at 121° C. for 30 minutes. After sterilization, 1 liter of sterile 50% dextrose (CERELOSE® 2001) and 500 ml of a 24 hours-grown inoculum were added. During the fermentation, the sugar was monitored but not adjusted. 
     After 15.5 hours of incubation, the dextrose and ethanol were determined and the optical density (OD) at 600 of a 1/100 dilution was measured. The cell density (1/100 dilution) reached an optical density of 0.215 at that time. A total of 175 grams of dry dextrose was added and allowed to mix for 15 minutes, after which 57.5 grams of acetophenone were added. Incubation was continued at 25° C., 300 rpm and 0.25 v/v/m aeration. Conversion was monitored by GC. The reaction taking place is as follows:                           
     D. Analysis: 
     Samples of broth were analyzed periodically by GC. 
     E. Product Recovery: 
     At the conclusion of the fermentation, the broth was pasteurized for 45 minutes at 65° C., cooled and stored in a refrigerator. The crude product is recovered by extraction of the whole broth using an equal volume of ethyl acetate. The spent broth is extracted a second time, and the recovered ethyl acetate layers are combined and washed two times with saturated aqueous NaCl. The solvent is then evaporated to about 8% using a rotary evaporator with a water bath temperature of 40° C. and a vacuum of 90-100 Torr. The crude extract is the fractionally distilled for use for its organoleptic properties as exemplified, infra; or it is used for conversion to styralyl acetate with or without any further purification. The resulting distilled product has a boiling point at atmospheric pressure of 204° C.; a melting point of 19.5-20.5° C.; a specific gravity of 15° C. of 1.013; and a refractive index at 20° C. of 1.5270. 
     NMR, GLC, chiral column GC and mass spectral analysis yield the information that the resulting product is a mixture of stereoisomers having the structures:                           
     having an optical rotation of −38.6°, [α−38.6°] and an enantiomeric excess percent of about 87.5, [εε%=87.5±0.5]. 
     EXAMPLE II 
     PRODUCTION OF STYRALYL ALCOHOL IN PILOT PLANT FERMENTERS 
     Reaction:                           
     I. Inoculum Preparation 
     A. Medium: 
     The following medium is prepared by admixing the following ingredients: 
     
       
         
               
               
             
               
             
           
               
                   
               
               
                 g/L 
                 Ingredients 
               
               
                   
               
             
             
               
                 5.0 
                 AMBERFERM ® 4000 Peptone 
               
               
                 5.0 
                 TASTONE ® 900 (yeast extract) 
               
               
                 5.0 
                 Malt extract 102 
               
               
                 5.0 
                 KH 2 PO 4  (potassium phosphate, monobasic) 
               
               
                 0.5 
                 MgSO 4 ·7H 2 O (magnesium sulfate, heptahydrate) 
               
               
                 1 L 
                 Deionized water 
               
               
                 1.0 
                 Antifoam Hi MARS-010 ® FGK 
               
               
                 50.0  
                 CERELOSE ® 2001 (added after sterilization) (dextrose) 
               
             
          
           
               
                 pH = 7.0, adjusted before sterilization with 25% aqueous NaOH solution 
               
               
                   
               
             
          
         
       
     
     B. Parameters: 
     
       
         
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 Temperature: 
                 25° C.; 
               
               
                   
                 Agitation: 
                 100 rpm; and 
               
               
                   
                 Duration: 
                 24 hours. 
               
               
                   
                   
               
             
          
         
       
     
     A Fernbach flask containing 1 liter of inoculum medium was sterilized for 30 minutes at 121° C. The flask was inoculated with 1.8 ml of a frozen culture of  Kluyveromyces polysporus  ATCC 22028; and 100 ml of sterile 50% dextrose solution were added. The flask was incubated in an incubator shaker (100 rpm) at 25° C. for 24 hours. 
     II. Seed Culture Preparation 
     A. Medium: 
     The following medium was prepared by admixing the following ingredients: 
     
       
         
               
               
             
               
             
           
               
                   
               
               
                 g/L 
                 Ingredients 
               
               
                   
               
             
             
               
                 100.0 
                 AMBERFERM ® 4000 Peptone 
               
               
                 100.0 
                 TASTONE ® 900 (yeast extract) 
               
               
                 100.0 
                 Malt extract 102 
               
               
                 100.0 
                 KH 2 PO 4  (potassium phosphate, monobasic) 
               
               
                  10.0 
                 MgSO 4 ·7H 2 O (magnesium sulfate, heptahydrate) 
               
               
                     20.0 L 
                 Deionized water 
               
               
                  10.0 
                 Antifoam Hi MARS-010 ® FGK 
               
               
                 1,000.0   
                 CERELOSE ® 2001 (added after sterilization) 
               
             
          
           
               
                 pH = 7.0, adjusted before sterilization with 25% NaOH 
               
               
                   
               
             
          
         
       
     
     B. Parameters: 
     
       
         
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 Broth Volume: 
                 20 liters; 
               
               
                   
                 Temperature: 
                 25° C.; 
               
               
                   
                 Agitation: 
                 100 rpm; 
               
               
                   
                 Aeration: 
                 0.1 v/v/m (2 liters/minute); 
               
               
                   
                 Duration: 
                 24 hours. 
               
               
                   
                   
               
             
          
         
       
     
     20 Liters of inoculum medium were sterilized for 30 minutes in a pilot plant 41 liter fermenter. After cooling, the fermenter was inoculated with 1 liter of cells grown for a period of 24 hours and 2 liters of sterile 50% dextrose were added. The fermenter was run at 25° C., 100 rpm and 0.1 v/v/m aeration for 24 hours. The pH is maintained in the range of from about 5.8 up to about 6. 
     III. Production 
     A. Medium: 
     The following medium was prepared by admixing the following ingredients: 
     
       
         
               
               
             
               
               
             
               
             
               
               
             
           
               
                   
               
               
                 g/L 
                 Ingredients 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 10.0 
                 AMBERFERM ® 4000 Peptone 
               
               
                 6.0 
                 TASTONE ® 900 (yeast extract) 
               
               
                 7.5 
                 Malt extract 102 
               
               
                 5.0 
                 KH 2 PO 4  (potassium phosphate, monobasic) 
               
               
                 0.5 
                 MgSO 4 ·7H 2 O (magnesium sulfate, heptahydrate) 
               
               
                      1.0 L 
                 Deionized water 
               
               
                 1.0 
                 Antifoam Hi MARS-010 ® FGK 
               
             
          
           
               
                 pH = 7.0, adjusted before sterilization with 25% NaOH 
               
             
          
           
               
                 56.6 
                 CERELOSE ® 2001 (added after sterilization) 
               
               
                 30.0 
                 CERELOSE ® 2001 (added after acetophenone) 
               
               
                 38.0 
                 Inoculum 
               
               
                 1.5 
                 NaOH (sodium hydroxide) 
               
               
                 5.0 
                 Acetophenone 
               
               
                 2.1 
                 Phosphoric acid 85% 
               
               
                   
               
             
          
         
       
     
     B. Parameters: 
     
       
         
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 Inoculum: 
                 20 liters of propagation culture; 
               
               
                   
                 Broth Volume: 
                 530 liters; 
               
               
                   
                 Temperature: 
                 25° C.; 
               
               
                   
                 Aeration: 
                 0.25 v/v/m; 
               
               
                   
                 Agitation: 
                 50 rpm; and 
               
               
                   
                 Duration: 
                 40 hours. 
               
               
                   
                   
               
             
          
         
       
     
     C. Process: 
     530 Liters of production medium were sterilized in the pilot plant 300 gallon fermenter for 30 minutes at 121° C. After sterilization and cooling, 60 liters of sterile 50% CERELOSE® dextrose and 20 liters of 24 hours-grown seed culture were added. The pH is maintained in the range, of from about 5.5 up to about 6. After 15.5 hours of incubation, when the optical density is at 600 of a 1/100 broth dilution was 0.171, a total of 15.9 kg dry dextrose powder was added. After 15 minutes of mixing, a total of 2.65 kg of acetophenone was added, and the agitation was increased to 100 rpm. 
     At the conclusion of the 24 hour incubation period, the broth is cooled for a period of 24 hours to room temperature and then pasteurized for 45 minutes at 65° C. The broth is then acidified to a pH in the range of 4-4.5 using 85% phosphoric acid, centrifuged and stored. 
     D. Product Recovery: 
     The broth is then extracted with ethyl acetate, and the ethyl acetate extract is evaporated. The resulting oil is then fractionally distilled at 204° C. and 1 atmosphere pressure to yield a product having a specific gravity of 1.013 and a refractive index at 20° C. of 1.5270. 
     The resulting product is a mixture of stereoisomers having the structures:                           
     having an optical rotation of 38.6°, [α=−38.6°] and an enantiomeric excess percent of about 87.5, [εε%=87.5 ±0.5]. 
     EXAMPLE III 
     PRODUCTION OF STYRALYL ACETATE 
     Reaction:                           
     Into a 2 liter reaction vessel equipped with stirrer, thermometer, reflux condenser and additional funnel are placed 118 grams of the crude: ethyl acetate extract of Example I, containing 85 grams of styralyl alcohol, a mixture of stereoisomers having the structures:                           
     212.5 grams of acetic acid and 0.3 grams of citric acid was refluxed for 15 hours. 
     During the refluxing, 168 grams of acetic acid were recovered. 
     The refluxing conditions are 118-120° C. at 5 mm/Hg pressure. 
     The total of 130 grams crude is recovered and fractionally distilled on a 2 inch×24 inch Goodloe column. Fractions the structures:                           
     having an optical rotation of−102.7°, [α=−102.7°] and an enantiomeric excess percent of about 78.31, [εε%=78.31±0.5]. 
     EXAMPLE IV 
     PREPARATION OF STYRALYL ACETATE 
     Reaction:                           
     Into a 500 ml flask equipped with stirrer, thermometer, reflux condenser and addition funnel are placed 134 grams of the styralyl alcohol mixture of stereoisomers prepared according to Example II; 0.5 grams water and 3.25 grams NOVOZYM® 435, an ester-forming enzyme which is  Candida antarctica  esterase expressed in  Asperigilus orzae , a triacylglycerol hydrolase (E.C. No. 3.1.1.3) acting as an effective carboxylesterase (trademark of the Novo Nordisk A/S Organization of Novo Alle, 2880 Bagsvaerd, Denmark). 
     The resulting mixture with stirring is heated to 40° C. and over a period of 8 hours at a rate of 7.5 grams per hour, 60 grams of acetic acid is added to the reaction mass. 
     The reaction mass is continued to be stirred for a period of 48 hours while maintaining the temperature at 40° C. 
     At the end of the 48 hour period, the resulting product is extracted with ethyl acetate and the ethyl acetate extract is concentrated and then fractionally distilled yielding a product, as confirmed by NMR, IR, mass spectral analysis and chiral GC analysis as a mixture of stereoisomers having the structures:                           
     having an optical rotation of +73.06°, [α=+73.06°] and an enantiomeric excess percent of about 81.1, [εε%=81.1±0.5]. 
     The resulting product has a boiling point at atmospheric pressure of 213° C., a specific gravity at 15° C. of 1.024 and a refractive index at 20° C. of 1.4948. 
     The conversion is 3.12%. 
     When the above example is carried out using in addition to the aforementioned reagents, 134 grams of n-hexane solvent, the conversion after 48 hours increases to 9.1%. 
     EXAMPLE V 
     CHYPRE FRAGRANCE FORMULATIONS 
     The following Chypre formulations are prepared: 
     
       
         
               
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                   
                   
               
               
                   
                 Parts by Weight 
               
             
          
           
               
                 Ingredients 
                 V(A) 
                 V(B) 
                 V(C) 
               
               
                   
               
             
          
           
               
                 Musk ambrette 
                 40 
                 40 
                 40 
               
               
                 Musk ketone 
                 60 
                 60 
                 60 
               
               
                 Coumarin 
                 30 
                 30 
                 30 
               
               
                 Oil of bergamot 
                 150 
                 150 
                 150 
               
               
                 Oil of lemon 
                 100 
                 100 
                 100 
               
               
                 Methyl ionone 
                 50 
                 50 
                 50 
               
               
                 Hexyl cinnamic aldehyde 
                 100 
                 100 
                 100 
               
               
                 Hydroxycitronellal 
                 100 
                 100 
                 100 
               
               
                 Oil of lavender 
                 50 
                 50 
                 50 
               
               
                 Texas cedarwood oil 
                 85 
                 85 
                 85 
               
               
                 Virginia cedarwood oil 
                 30 
                 30 
                 30 
               
               
                 Oil of sandalwood (East Indies) 
                 40 
                 40 
                 40 
               
               
                 Isoeugenol 
                 20 
                 20 
                 20 
               
               
                 Eugenol 
                 10 
                 10 
                 10 
               
               
                 Benzyl acetate 
                 30 
                 30 
                 30 
               
               
                 beta-Phenyl ethyl alcohol 
                 40 
                 40 
                 40 
               
               
                 alpha-phenyl ethyl alcohol 
                 30 
                 30 
                 30 
               
               
                 Oakmoss absolute 
                 30 
                 30 
                 30 
               
               
                 Vetiver oil of Venezuela 
                 25 
                 25 
                 25 
               
               
                 The mixture of stereoisomers defined according 
                 25 
                 0 
                 0 
               
               
                 to the structures: 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                   
               
               
                 prepared according to Example I, optical rotation 
               
               
                 −38.6°, [α = −38.6°] with an enantiomeric 
               
               
                 excess percent of 
               
               
                 about 87.5, [εε% = 87.5 ± 0.5]. 
               
               
                 The mixture of stereoisomers having the structures: 
                 0 
                 25 
                 0 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                   
               
               
                 prepared according to Example III, having an optical 
               
               
                 rotation of −102.7°, [α = −102.7°] with an 
               
               
                 enantiomeric excess percent of about 
               
               
                 78.31, [εε% = 78.31 ± 0.5]. 
               
               
                 The mixture of stereoisomers prepared according 
                 0 
                 0 
                 25 
               
               
                 to Example IV having the structures: 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                   
               
               
                 having an optical rotation of 73.06°, 
               
               
                 [α = +73.06°] with 
               
               
                 an enantiomeric excess percent of about 81.1, 
               
               
                 [εε% = 81.1 ± 0.5]. 
               
               
                   
               
             
          
         
       
     
     The mixture of styralyl alcohol stereoisomers prepared according to Example I imparts to this Chypre formulation hyacinth, gardenia undertones with strawberry topnotes. Accordingly, the aroma of the Chypre formulation of Example V(A) is described as: “a Chypre aroma with hyacinth and gardenia undertones and strawberry topnotes.” 
     The mixture of styralyl acetate stereoisomers of Example III imparts to this Chypre formulation fresh strawberry, green, dried fruit undertones with green, strawberry topnotes. Accordingly, the aroma of the Chypre formulation of Example V(B) is described as: “a Chypre aroma with strawberry, green and dried fruit undertones and green, strawberry topnotes.” 
     The mixture of styralyl acetate stereoisomers of Example IV imparts to the Chypre formulation of Example V(C) fruity, floral, mimosa, jasmine and gardenia undertones with fruity, strawberry topnotes. Accordingly, the aroma of the Chypre formulation of Example V(C) is described as: “a Chypre aroma with fruity, floral, mimosa, jasmine and gardenia undertones and strawberry, fruity topnotes.” 
     EXAMPLE VI 
     PREPARATION OF A COSMETIC POWDER COMPOSITION 
     Cosmetic powder compositions are prepared by nixing in a ball mill 100 grams of talcum powder with 0.25 grams of each of the substances set forth in Table II below. Each of the cosmetic powder compositions has an excellent aroma as described in Table II below. 
     
       
         
               
               
             
           
               
                 TABLE II 
               
               
                   
               
               
                 Substance 
                 Aroma Description 
               
               
                   
               
             
             
               
                 The mixture of stereoisomers having the 
                 A hyacinth, gardenia aroma with 
               
               
                 structures: 
                 strawberry topnotes. 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                   
               
               
                 prepared according to Example I, 
               
               
                 having an optical rotation of 
               
               
                 −38.60, [α = −38.6°  with an 
               
               
                 enantiomeric excess percent of 
               
               
                 about 87.5, 
               
               
                 [εε% = 87.5 ± 0.5]. 
               
               
                 The mixture of stereoisomers having 
                 A strawberry, green, dried fruit 
               
               
                 the structures: 
                 aroma with green, strawberry 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 topnotes. 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                   
               
               
                 prepared according to Example III, 
               
               
                 having an optical rotation of 
               
               
                 −102.7, [α = −102.7°  with an 
               
               
                 enantiomeric excess percent of 
               
               
                 about 78.31, 
               
               
                 [εε% = 78.31 ± 0.5]. 
               
               
                 A mixture of stereoisomers having 
                 A fruity, floral, jasmine, 
               
               
                 the structures: 
                 mimosa, gardenia aroma with 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 fruity and strawberry topnotes. 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                   
               
               
                 having an optical rotation of +73.60°, 
               
               
                 [α = +73.06°] with an 
               
               
                 enantiomeric excess percent of about 
               
               
                 81.1, [εε% = 81.1 ± 0.5]. 
               
               
                 prepared according to Example IV. 
               
               
                 The perfume composition of 
                 A Chypre aroma with hyacinth 
               
               
                 Example V(A). 
                 and gardenia undertones and 
               
               
                   
                 strawberry topnotes. 
               
               
                 The perfume composition of 
                 A Chypre aroma with 
               
               
                 Example V(B). 
                 strawberry, green and dried 
               
               
                   
                 fruit undertones and green, 
               
               
                   
                 strawberry topnotes. 
               
               
                 The perfume composition of 
                 A Chypre aroma with fruity, 
               
               
                 Example V(C). 
                 floral, mimosa, jasmine and 
               
               
                   
                 gardenia undertones and 
               
               
                   
                 strawberry, fruity topnotes. 
               
               
                   
               
             
          
         
       
     
     EXAMPLE VII 
     PERFUMED LIQUID DETERGENT 
     Concentrated liquid detergents (lysine salt of n-dodecylbenzene sulfonic acid as more specifically described in U.S. Pat. No. 3,948,818 issued on Apr. 6, 1976, incorporated by reference herein) with aroma nuances as set forth in Table II of Example VI are prepared by adding 0.10%, 0.15%, 0.20%, 0.25%, 0.30% and 0.35% of the substance set forth in Table II of Example VI. They are prepared by adding and homogeneously mixing the appropriate quantity of substances set forth in Table II of Example VI in the liquid detergent. The detergents all possess excellent aromas as set forth in Table II of Example VI, the intensity increasing with greater concentrations of substance as set forth in Table II of Example VI. 
     EXAMPLE VIII 
     PREPARATION OF A COLOGNES AND HANDKERCHIEF PERFUMES 
     Compositions as set forth in Table II of Example VI are incorporated into colognes at concentrations of 2.0%, 2.5%, 3.0%, 3.5%, 4.0% and 5.0% in 80%, 85%, 90% and 95% aqueous: food grade ethanol solutions; and into handkerchief perfumes at concentrations of 15%, 20%, 25% and 30% (in 80%, 85%, 90% and 95% aqueous food grade ethanol solution). Distinctive and definite fragrances as set forth in Table II of Example VI are imparted to the colognes and to the handkerchief perfumes at all levels indicated. 
     EXAMPLE IX 
     PREPARATION OF A SOAP COMPOSITION 
     100 Grams of soap chips (per sample) (IVORY®, produced by the Procter &amp; Gamble Company of Cincinnati, Ohio) are each mixed with 1 gram samples of substances as set forth in Table II of Example VI until homogenous compositions are obtained. In each of the cases, the homogeneous compositions are heated under 8 atmospheres pressure at 180° C. for a period of 3 hours and the resulting liquids are placed into soap molds. The resulting soap cakes, on cooling, manifest aromas as set forth in Table II of Example VI. 
     EXAMPLE X 
     PREPARATION OF SOLID DETERGENT COMPOSITIONS 
     Detergents are prepared using the following ingredients according to Example I of Canadian Patent No. 1.007,948 (incorporated by reference herein): 
     
       
         
               
               
             
           
               
                   
               
               
                 Ingredients 
                 Parts by Weight 
               
               
                   
               
             
             
               
                 NEODOL ® 45-11 (a C 14 -C 15  alcohol ethoxylated 
                 12 
               
               
                 with 11 moles of ethylene oxide) 
               
               
                 Sodium carbonate 
                 55 
               
               
                 Sodium citrate 
                 20 
               
               
                 Sodium sulfate, water brighteners 
                 q.s. 
               
               
                   
               
             
          
         
       
     
     The detergent is a phosphate-free detergent. Samples of 100 grams each of this detergent are admixed with 0.10, 0.15, 0.20 and 0.25 grams of each of the substances as set forth in Table II of Example VI. Each of the detergent samples has an excellent aroma as indicated in Table II of Example VI. 
     EXAMPLE XI 
     Utilizing the procedure of Example I at column 15 of U.S. Pat. No. 3,632,396 (the disclosure of which is incorporated by reference herein), a non-woven cloth substrate useful as drier-added fabric softening articles of manufacture are prepared wherein the substrate, substrate coating, outer coating and the perfuming material are as follows: 
     (1) a water “dissolvable” paper (“Dissolve Paper”): 
     (2) ADOGEN® 448 (melting point about 140° F.) as the substrate coating; and, 
     (3) an outer coating having the following formulation (melting point about 150° F.). 
     57% C 20 -C 22  HAPS; 
     22% isopropyl alcohol; 
     20% antistatic agent; and 
     1% of one of the perfume materials as set forth in Table II of Example Vl. 
     Fabric softening compositions, prepared according to Example I at column 15 of U.S. Pat. No. 3.632,396 having aroma characteristics as set forth in Table II of Example VI, consist of a substrate coating having a weight of about 3 grams per 100 square inches of substrate: a first coating on the substrate coating consisting of about 1.85 grams per 100 square inches, of substrate; and an outer coating coated on the first coating consisting of about 1.4 grams per 100 square inches of substrate. One of the substances of Table II of Example VI is admixed in each case with the outer coating mixture, thereby, providing a total aromatized outer coating weight ratio to substrate of about 0.5:1 by weight of the substrate. The aroma characteristics are imparted in a pleasant manner to the headspace in a drier on operation thereof in each case, using the said drier-added fabric softener, non-woven fabrics and these aroma characteristics are described in Table II of Example VI. 
     EXAMPLE XI 
     HAIR SPRAY FORMULATIONS 
     The following hair spray formulation is prepared by first dissolving PVP/NA E-735 copolymer manufactured by the GAF Corporation of 140 West 51st Street, New York, N.Y. in 91.62 grams of 95% food grade ethanol. 8.0 Grams of the polymer is dissolved in the alcohol. The following ingredients are added to the PVP/NA alcoholic solution: 
     
       
         
               
               
             
           
               
                   
               
               
                 Ingredients 
                 Weight Percent 
               
               
                   
               
             
             
               
                 Dioctyl sebacate 
                 0.05 
               
               
                 Benzyl alcohol 
                 0.10 
               
               
                 Dow Corning 473 fluid prepared by the Dow Corning 
                 0.10 
               
               
                 Corporation 
               
               
                 TWEEN ® 20 surfactant (prepared by ICI 
                 0.03 
               
               
                 America Corporation) 
               
               
                 One of the perfumery substances as set forth in Table II 
                 0.10 
               
               
                 of Example VI. 
               
               
                   
               
             
          
         
       
     
     The perfuming substances as set forth in Table II of Example VI add aroma characteristics as set forth in Table II of Example VI, which are rather intense and aesthetically pleasing to the users of the soft-feel, good-hold pump hair sprays. 
     EXAMPLE XIII 
     CONDITIONING SHAMPOOS 
     Monamid CMS (prepared by the Mona Industries Company) (3.0 weight percent) is melted with 2.0 weight percent coconut fatty acid (prepared by the Procter &amp; Gamble Company of Cincinnati, Ohio); 1.0 weight percent ethylene glycol distearate (prepared by the Armak Corporation); and triethanolamine (a product of the Union Carbide Corporation). (1.4 weight percent). The resulting melt is admixed with Stepanol WAT produced by the Stepan Chemical Company, (35.0 weight percent). The resulting mixture is heated to 60° C. and mixed until a clear solution is obtained (at 60° C.). 
     GAFQUAT® 755N polymer (manufactured by GAF Corporation of 140 West 51st Street, New York, N.Y.) (5.0 weight percent) is admixed with 0.1 weight percent sodium sulfite and 1.4 weight percent polyethylene glycol 6000 distearate produced by the Armak Corporation. 
     The resulting material is then mixed and cooled to 45° C., and 0.3 weight percent of perfuming substance as set forth in Table II of Example VI is added to the mixture. The resulting mixture is cooled to 40° C., and blending is carried out for an additional 1 hour in each case. At the end of this blending period, the resulting material has a pleasant fragrance as indicated in Table II of Example VI. 
     EXAMPLE XIV 
     The following three tapioca flavoring formulations are prepared: 
     
       
         
               
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                   
                   
               
               
                   
                 Parts by Weight 
               
             
          
           
               
                 Ingredients 
                 XIV(A) 
                 XIV(B) 
                 XIV(C) 
               
               
                   
               
             
          
           
               
                 2-Methyl-2-pentenoic acid 
                 5 
                 5 
                 5 
               
               
                 2-Methyl-3-pentenoic acid 
                 8 
                 8 
                 8 
               
               
                 Ethyl ester of 2-methyl-3-pentenoic acid 
                 4 
                 4 
                 4 
               
               
                 n-Hexyl ester of 2-methyl-4-pentenoic acid 
                 6 
                 6 
                 6 
               
               
                 The mixture of stereoisomers having 
                 2.5 
                 0 
                 0 
               
               
                 the structures: 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                   
               
               
                 prepared according to Example I, having an 
               
               
                 optical rotation −38.6°, 
               
               
                 [α = −38.6°] with an enantiomeric excess 
               
               
                 percent of about 87.5, [εε% = 87.5 ± 0.5]. 
               
               
                 The mixture of stereoisomers having the 
                 0 
                 2.5 
                 0 
               
               
                 structures: 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                   
               
               
                 prepared according to Example III, having an 
               
               
                 optical rotation of −102.7°, 
               
               
                 [α = −102.7°] with an enantiomeric excess 
               
               
                 percent of about 78.31, [εε% = 78.31 ± 0.5]. 
               
               
                 The mixture of stereoisomers prepared 
                 0 
                 0 
                 2.5 
               
               
                 according to Example IV having the 
               
               
                 structures: 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                   
               
               
                 having an optical rotation of +73.06°, 
               
               
                 [α = +73.06°] with an enantiomeric excess 
               
               
                 percent of about 81.1, [εε% = 81.1 ± 0.5]. 
               
               
                   
               
             
          
         
       
     
     At the rate of 11 ppm, each of the above strawberry formulations are added to separate portions of tapioca pudding formulations, and the tapioca pudding formulations are crafted into tapioca pudding portions. 
     The tapioca pudding portion of Example XIV(A) has an excellent strawberry taste with fragrant strawberry jam aromas. 
     The tapioca pudding portion of Example XIV(B) has an excellent green, fresh strawberry taste with green, avocado and strawberry jam nuances. 
     The tapioca pudding portion of Example XIV(C) has an excellent strawberry jam taste with apricot and apple flavor nuances.