METHOD FOR OBTAINING FOOD COLOURING PRODUCT

Abstract: According to this method colourant for foodstuffs is obtained. Red and orange pigments obtained as fungi belonging to genus Monascus are cultivated on culture media, based on milk or whey permeate with different carbon and nitrogen sources and microelements added. Through this method a complex, ecologically clean product, containing lactose, protein, amino acids, vitamins and water soluble pigment is obtained. The method is wasteless, ecologically clean and a nontraditional substrate, waste from the milk industry is utilized.

Description: METHOD FOR OBTAINING OF FOOD COLOURING PRODUCT FIELD OF INVESTIGATION The invention reffers to method for obtaining of a colouring agent for foodstuffs, which can find application in the food and beverage industry. BACKGROUND OF THE INVENTION The fungi, belonging to genus Monascus are well known as natural colouring agents in the food and beverage industry. They have been used in the oriental countries since centuries, as main colouring agent of wine, meat and curd (1) . Usually the pigments are obtained as watter or oil soluble substances stable in the pH region 2 - 10, and at high teperatures (120"C) (2) . Different methods used for cultivation of these fungi for pigment biosynthesis are known. They are characterized with the utilization of solid and liquid complex substrates for growth and solid phase and submerged cultivation (3.4) . Natural substrates for the fungi are different cereals and especially rice, and after that the red mixuture is grained and applied as powder (5) . Disadvantages of these methods are the long duration (4 - 18 days) , long step procedure and obtaining only a pigmented product. TECHNICAL BACKGROUND The aim of the invention is to create a method for obtaining of a complex colouring agent for foodstuffs. The aim of the invention can be achieved by using strain microorganism producer of pigments, belonging to genus Monascus . The following strains are used: Monascus purpureus CBS 109. 07, Monascus purpureus DSM 1604, Monascus pu pure us DSM 1603, Monascus ruber DSM 62748, Monascus anka IFO 6540, Monascus anka IFO 4478, Monascus purpureus IFO 4513. As culture medium, milk whey and/or whey permeate and milk permeate in concentration 10 - 100 % , pH 3 - 7, on which the above mentioned m croorganisms can grow and produce pigments is used. To this main culture medium different substances can be added: additional carbon sources 0.1 - 5?6, natural starch sources 0.5 - 5 %, inorganic nitrogen sources 0.1 - 1 % , organic nitrogen sources 0.1 - 10 % , mineral salts 0.001 - 1 % . It was found that the use of whey or milk permeate with different, carbon sources show sinergic effect on the Monascus pigment production. As additional carbon sources sugars, aminosugars, sugar alcohols, alcochols and natural sources containing starch are used. As additional nitrogen sources amonium and nitrate salts, organic nitrogen containing substances as yeast extract, malt extract, corn extract etc. can be added. Additionally mineral salts like phosphates, sulphates, sodium. magnesium and potassium salts can be used. The cultivation methods applied in this invention are submerged - batch, fed - batch, continuous. The microorganisms from genus Monascus are cultivated at temperature 25 - 40°C for 2 - 5 days. The fermentation process is performed under the following conditions: pH 4 - 8, air 0.3 - 1 1/1/min, agitation 200 - 500 r.p. ., p0 2 20 - 50°ό. For the pigment isolation from the cultural broth all known methods can be used. For example the culture can be applied as a colouring agent in liquid form in the foodstuffs manufacture or as freeze - dried or spray - dried product. In some cases the mycellium can be removed by filtration techniques, and when necessary the liquid product can be concentrated up to 40 - 50 % dry weight by vacuum evaporation. The advantages of the method described in this invention can be summarized as follows: a submerged, one stage method is used, the cultivation process is short; batch and fed - batch cultivation methods are used; complex and ecologically clean product is obtained, which except pigment contains lactose, proteins, amino acids, vitamins and mineral salts; the red pigment is totally water soluble while the orange pigment is in crystaline form. As main culture medium a cheap, non - traditional substrate. waste from the milk industry, is used and through this method a very strong pollutant from the milk industry is converted into a valuable product for food and beverage industry. The method is ecologically clean, the technology is wastless. EXAMPLE 1 The Monascus purpureus DSM 1604 strain is maintained on solid medium containing (g/1) : malt extract 5, peptone 10, glucose 20, agar 20, pH 6.0. The cultivation is carried out at 30°C for 7 days. The main fermentation is performed in a 10 1 bioreactor. The strain is cultivated on culture medium ultrafi ltrate of milk whey with decreased protein content. The culture medium is prepared in the following way: cheese or yellow cheese whey is clarified by the following procedure: to 10 1 whey Ca.C l __ in quantity up to lg/kg is added ph is justed to 7.5 with KOH and after that it is heated to 50 - 60 * C for 15 min. As a result of this procedure part of the fat content is removed and after that an ultrafi tration on DDS equipment with 5 kDa membranes is carried out. The permeate obtained contains (g/1) lactose 45 - 50, protein 0.9 - 1, nitrogen 1.0 - 1.3, phosphorus 2.0 - 2.5, is used as a medium for the fungus cultivation. The cultivation of the strain producer is carried out under the following conditions: temperature 30°C, pH 6.5, air 0.3 1/1/min, agitation 200 r.p.m. The cultivation lasts 72 h and the obtained product is separated from the biomass by centrifugation. The sugar (6), nitrogan (7), phosphorus (8), protein (9), pigment (10), dry weight (11) contain and the proteolitic activity (12) are determined. The filtrate is freeze dried. In the case when the proteolitic activity is not desired a heating procedure at 120 , 'C for 15 min is involved. The product obtained possess the following characteristics: powderlike, red, watersoluble substance, containing (mg/g dry product) : lactose 970, pigment 180 - 200 OU, protein 16.8, proteolitic activity 1 U/mg protein. EXAMPLE 2 Monascus purpureus CBS 109. 07 is used. The cultivation is carried out under the conditions described in EXAMPLE 1. The culture medium is milk permeate, diluted 1:1 and the following ingradients are added (g/1) : yeast extract 0.5, KH 2 P0J4 1.0, MgSO., 7H 2 0 0.5, ZnS0 4 7H z O 0.001, pH 6.0. As additional carbon sources different sugars, aminosugars and alchocols are used. The results are shown on Table 1. At the end of the cultivation process, the cultural broth is spray dried. The product obtained possesses the following characteristics: powderlike, red, water soluble substance, containing pigment 250 - 550 OU / mg dry weight (Table 1) . The carbon sources added, do not influence the physical properties of the product obtained. abl EXAMPLE 3 Monascus ruber DSM 62748 is used. Cultivation is performed under the conditions described in EXAMPLE 2 and as additional carbon source glucose in concentration 2 ?ό is added. As nitrogen sources different substances are added to the culture medium (Table 2) . The product obtained after spray drying of the fermentation broth possess the following characteristics: powderlike, red coloured watersoluble substance with pigment content 200 - 500 OU/mg dry weight and powderlike, orange substance with pigment contain 300 - 600 OU/mg dry weight (Table 2) . Table 2. EXAMPLE 4 Monascus anka IFO 4478 strain is used. The cultivation is carried out under the conditions described in EXAMPLE 1. The culture medium used is milk permeate 50 % with additional carbon substrates different natural starch sources in concentration 1 % (Table3) . The product obtained after spray drying possess the following characteristic: powderlike, red coloured, watersoluble substance, with pigment contain 500 - 600 OU/mg dry weight (Table 3) . Table 3. REFERENCES 1. Wong H.- C; Koehler P.E., Production and isolation of antibiotic from Monascus purpureus and its relationship to pigment production, 1981, J.Food Sci . 46, 8, 589 2. JP 60 149 512, Make up containing natural olours, abstract 10374 in Chemical Abstracts, vol 104. 3. US Patent 4, 927, 760 Production of crystaline pigment from Monascus during fermentation 4. US Patent 4, 145, 254 Red pigment production 5. US Patent 4, 418, 080 Natural red colouring prepared from wheat and barley substrates 6 Somogyi M.J. Notes on sugar determination. J. Biol. Chem. 1952, 195, 19. 7. Jacobs. S., Meth. biochem. Anal., 13, 241, 196. . 8. King E.J..Biochem. J. 26. 292, 1932. 9. Lowry O.H., Rosenbrought N.J. Farel O.M., Randal V.J., J. Biol. Chem. 1951, 193, 265. 10. Su Y.C.; Huang J.H., Fermentative Production of anka pigments, 1980, Proc. Nat1. Sci. Counc. ROC, 4, 2, 201. 11. Bau Y.S. ; Wong H.C., Z nk effec-: on growth, pigmentation and antibacterial activity of Monascus purpureus, 1977, Physiol . Plants, 46, 63. 12. Applied Biochemistry and Microbiology, 1973, ed pu l . house Mir, Moscow, T. 7, 256.

Claims: CLAIMS 1. Method for obtaining a colorant from pigment synthesizing fungi, belonging to genus Monascus with application in the food and beverage industry. It is characterized as follows: the microorganisms are cultivated in a submerged way at temperature 25 - 40°C, pH 4 - 7, air 0.3 - 1 1/1/min, agitation 200 - 500 r.p.m. for 2 - 5 days on culture medium on the basis of 10 - 100 % secondary milk products with 0.1 - 5 % carbon and 0.1 - 3% nitrogen sources, 0.1 - 1% mineral salts added, and the pigment obtained is isolated. 2. Method according claim 1, characterized with that as pigment producers the following strains from fungi belonging to genus Monascus are used Monascus purpureus CBS 109. 07, Monascus purpureus DSM 1604, Monascus purpureus DSM 1603, Monascus purpureus IFO 4513, Monascus ruber DSM 62748, Monascus anka IFO 6540, Monascus anka IFO 4478. 3. Method according claim 1, characterized with that as secondary milk products milk whey, whey permeate and m lk permeate are used. 4. Method according claim 1, characterized with that as additional carbon source sugars, aminosugars, sugar alcohols and alcohols (0.1 - 5%) and natural sources of starch (0.5 - 3%) are added. 5. Method according claim 1, characterized with that to the main culture medium additional nitrogen sources - ammonium, nitrates, yeast, malt, corn extract etc. in concentration 0.1 -3% are added. 6. Method according claim I, characterized with that as additional mineral salts potassium, sodium, magnesium, sulphate and phosphate salts are added. 7. Method according claim 1, characterized with that the product obtained is isolated by applying membrane filtration, extraction with organic solvents or the cultivation broth is vaccuum concentrated, spray dried or freeze dried.

Inventors: DAVIDOVA EUGENIA, KUJUMDZIEVA ANNA, RASHEVA TANIA, SAVOV VALENTIN, TODOROV TZONI, WARTENBERG ARNOLD, DAVIDOVA, EUGENIA, KUJUMDZIEVA, ANNA;, RASHEVA, TANIA;, SAVOV, VALENTIN;, TODOROV, TZONI;, WARTENBERG, ARNOLD
Applicants: DAVIDOVA EUGENIA, KUJUMDZIEVA ANNA, RASHEVA TANIA, SAVOV VALENTIN, TODOROV TZONI, WARTENBERG ARNOLD, DAVIDOVA, EUGENIA, KUJUMDZIEVA, ANNA;, RASHEVA, TANIA;, SAVOV, VALENTIN;, TODOROV, TZONI;, WARTENBERG, ARNOLD
Application year: 1993
Main classification: C12P1
IPC codes: C09B61, C12P1, C12P17