WO2022166398A1 - Cysteine-rich yeast extract and preparation method therefor - Google Patents

Cysteine-rich yeast extract and preparation method therefor Download PDF

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Publication number
WO2022166398A1
WO2022166398A1 PCT/CN2021/137074 CN2021137074W WO2022166398A1 WO 2022166398 A1 WO2022166398 A1 WO 2022166398A1 CN 2021137074 W CN2021137074 W CN 2021137074W WO 2022166398 A1 WO2022166398 A1 WO 2022166398A1
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yeast
yeast extract
enzymatic hydrolysis
cysteine
culture
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PCT/CN2021/137074
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French (fr)
Chinese (zh)
Inventor
刘向军
熊涛
唐冠群
李库
颜东方
胥怀
廖辉
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安琪酵母股份有限公司
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Publication of WO2022166398A1 publication Critical patent/WO2022166398A1/en

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/20—Synthetic spices, flavouring agents or condiments
    • A23L27/21—Synthetic spices, flavouring agents or condiments containing amino acids
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/20—Synthetic spices, flavouring agents or condiments
    • A23L27/26—Meat flavours
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
    • A23L31/10—Yeasts or derivatives thereof
    • A23L31/15—Extracts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C12N1/18—Baker's yeast; Brewer's yeast
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi
    • C12R2001/85—Saccharomyces
    • C12R2001/865—Saccharomyces cerevisiae

Definitions

  • the invention relates to the field of yeast extracts, in particular to a cysteine-rich yeast extract and a preparation method thereof.
  • Yeast extract is a natural seasoning with food properties, which is made of edible yeast as raw material and refined by degrading proteins and nucleic acids in yeast cells. It contains proteins, polypeptides, amino acids, nucleotides, etc.
  • the active ingredient has the characteristics of freshening, thickening and lasting aftertaste, and is widely used in the condiment industry.
  • cysteine a precursor for providing meaty taste.
  • cysteine content in the yeast extract prepared in the prior art is relatively low, generally below 1%.
  • the prior art discloses a method for converting cystine to cysteine and/or glutathione, comprising contacting cystine with microorganisms. Also disclosed are yeast extracts comprising at least 1.8 mg/g cysteine and yeast autolysates comprising at least 1.3 mg/g cysteine.
  • the cysteine content is 1.8mg/g (ie, 1.8 ⁇ ), which is much lower than 1%.
  • the technical problem solved by the present invention is that the cysteine content of yeast autolysate or yeast extract in the prior art is relatively low, far below 1%.
  • the invention provides a yeast extract with a cysteine content of more than 1% and a preparation method thereof.
  • the present invention provides the following technical solutions:
  • the present invention provides a cysteine-rich yeast extract prepared by comprising the following steps:
  • step (2) the yeast milk obtained in step (1) is thermally extracted and centrifuged to obtain a supernatant;
  • step (3) enzymolysis of the supernatant obtained in step (2) to obtain a cysteine-rich yeast extract
  • the original strain described in the step (1) is a yeast strain (saccharomyces cerevisiae) rich in reduced glutathione, the deposit number is: CCTCC M 205130, and the deposit address is: on October 25, 2005, it was preserved in China Type Culture Collection (Wuhan).
  • the cysteine content in the yeast extract is greater than or equal to 1%, preferably the cysteine content in the yeast extract is greater than or equal to 2% based on the mass of the yeast extract dry matter.
  • the fermentation process described in step (1) adds nutrient solution in a fed-feed mode; preferably, the fermentation process includes an ordinary culture process and a starvation culture process, and in percentage by weight, the nutrient solution added flow rate in the starvation culture process Add 10-40% of the flow rate for normal culture procedures.
  • the temperature of the normal culture is 25-35°C, and the time of the normal culture is preferably 5-20h; further preferably, the temperature of the starvation culture is 25-35°C, and the time of the starvation culture is preferably 5-10h .
  • the mass concentration of the yeast milk used in the thermal extraction in step (2) is 8-15%; the temperature of the thermal extraction is preferably 80-110°C, more preferably 95°C; the pH value during extraction is preferably 5.0-8.0; The extraction time is 60-120min, more preferably 90min.
  • endoprotease and exoprotease are used for enzymolysis in sequence, preferably the endoprotease is selected from one of alkaline protease, papain, trypsin and neutral protease or two or more; further preferably, the exoprotease is selected from one or more of flavor protease, carboxypeptidase and aminopeptidase.
  • the added amount of the endoprotease is 0.1-1.5%, preferably 1%, based on the mass of the dry matter of the supernatant.
  • the enzymatic hydrolysis temperature is 40-80° C., preferably, the enzymatic hydrolysis pH value is 4.0-7.5, and the enzymatic hydrolysis time is 10-15 h.
  • the amount of the exoprotease added is 0.1-1.5%, preferably 1%, based on the mass of the dry matter of the supernatant.
  • the enzymatic hydrolysis temperature is 40-80° C.; preferably, the enzymatic hydrolysis pH value is 4.0-7.5, and the enzymatic hydrolysis time is 5-10 h.
  • step (3) before the enzymatic hydrolysis in step (3), it further comprises: adding vitamin C to the supernatant obtained in step (2); 2%; more preferably 1%.
  • step (3) it also includes the following steps: after step (3), it also includes the following steps: the enzymatic hydrolysis solution after the enzymatic hydrolysis in step (3) is successively evaporated, concentrated and dried to obtain cysteine-rich of yeast extract.
  • the drying method is one of spray drying, vacuum drying or drum drying, more preferably spray drying, and even more preferably, the inlet air temperature of spray drying is controlled at 155-175°C, and the outlet air temperature is controlled at 90-115°C °C, the atomization pressure is controlled at 100-160bar, and the receiving temperature is controlled at 25-45°C.
  • the invention provides a preparation method of a cysteine-rich yeast extract, which specifically comprises the following steps:
  • step (2) the yeast milk obtained in step (1) is thermally extracted and centrifuged to obtain a supernatant;
  • step (3) Enzymatic hydrolysis of the supernatant obtained in step (2) to obtain a cysteine-rich yeast extract.
  • the raw materials used in the present invention are non-toxic and harmless edible raw materials, and the prepared products and by-products can be used in the food industry;
  • the preparation method provided by the present invention has a high extraction rate, and the cysteine content of the product yeast extract is high, up to 2%.
  • the present invention provides a cysteine-rich yeast extract prepared by a preparation method comprising the following steps:
  • step (2) the yeast milk obtained in step (1) is thermally extracted and centrifuged to obtain a supernatant;
  • step (3) enzymolysis of the supernatant obtained in step (2) to obtain a cysteine-rich yeast extract
  • the original strain described in the step (1) is a yeast strain (saccharomyces cerevisiae) rich in reduced glutathione, the deposit number is: CCTCC M 205130, and the deposit address is: on October 25, 2005, it was preserved in China Type Culture Collection (Wuhan).
  • the cysteine content in the yeast extract is greater than or equal to 1% based on the mass of the dry matter of the yeast extract, preferably the cysteine content in the yeast extract is greater than or equal to 1%. 2%.
  • the fermentation process described in step (1) adds nutrient solution in a fed-feed way;
  • the addition flow of the nutrient solution in the starvation culture process is 10-40% of that in the ordinary culture process.
  • the nutrient solution includes the following weight percentages: 1% of yeast extract, 2% of peptone and 2% of glucose.
  • the temperature of the ordinary culture is 25-35°C, and preferably the time of the ordinary culture is 5-20h; further preferably, the temperature of the starvation culture is 25-35°C, preferably starvation The incubation time is 5-10h.
  • the mass concentration of the yeast milk used in the thermal extraction in step (2) is 8-15%; the temperature of the thermal extraction is preferably 80-110°C, more preferably 95°C; the pH value during extraction is preferably is 5.0-8.0; the preferred thermal extraction time is 60-120min, more preferably 90min; after the thermal extraction, the thermally extracted product is subjected to centrifugal treatment to remove impurities to obtain a supernatant, wherein the centrifugal speed is 5000rpm, and the centrifugal time is 20 minutes .
  • endoprotease and exoprotease are used for enzymolysis in sequence, preferably the endoprotease is selected from the group consisting of alkaline protease, papain, trypsin and intermediate protease. one or two or more of the protease; further preferably, the exoprotease is selected from one or more of flavor protease, carboxypeptidase and aminopeptidase.
  • the added amount of the endoprotease is 0.1-1.5%, preferably 1%, based on the mass of the supernatant dry matter.
  • the enzymatic hydrolysis temperature is 40-80° C., preferably, the enzymatic hydrolysis pH value is 4.0-7.5, and the enzymatic hydrolysis time is 10-15 h.
  • the amount of the exoprotease added is 0.1-1.5%, preferably 1%, based on the mass of the supernatant dry matter.
  • the enzymatic hydrolysis temperature is 40-80° C.; preferably, the enzymatic hydrolysis pH value is 4.0-7.5, and the enzymatic hydrolysis time is 5-10 h.
  • enzymatic hydrolysis further comprises: adding vitamin C to the supernatant obtained in step (2);
  • the addition amount of C is 0.5-2%; more preferably, it is 1%.
  • the yeast extract solution after enzymatic hydrolysis in step (3) is heated to 80° C. for 2 hours to inactivate enzymes, and evaporated and concentrated to obtain a concentrated product,
  • the solid content of the concentrate is 35-45%;
  • the concentrate is dried to obtain a cysteine-rich yeast extract.
  • the drying method is one of spray drying, vacuum drying or drum drying, more preferably spray drying, and even more preferably, the inlet air temperature of spray drying is controlled at 155-175°C, and the outlet air temperature is controlled at 90-115°C , the atomization pressure is controlled at 100-160bar, and the receiving temperature is controlled at 25-45 °C.
  • the invention provides a preparation method of a cysteine-rich yeast extract, which specifically comprises the following steps:
  • step (2) the yeast milk obtained in step (1) is thermally extracted and centrifuged to obtain a supernatant;
  • step (3) Enzymatic hydrolysis of the supernatant obtained in step (2) to obtain a cysteine-rich yeast extract.
  • the cysteine-rich yeast extract and the preparation method thereof of the present invention will be further described below through specific examples.
  • the present invention adopts the yeast strain rich in reduced glutathione ((saccharomyces cerevisiae), the deposit number is: CCTCC M 205130, and the deposit address is: on October 25, 2005, it was deposited in the China Center for Type Culture Collection (Wuhan) The strain has been recorded in the patent publication with publication number CN101575578A.
  • the dry matter content of yeast milk and the dry matter content of the supernatant are measured by the following drying weighing method:
  • the yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast paste, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by feeding feed flow.
  • the culture time was 20h; then starvation culture was carried out, and the nutrient solution was added by the feeding method, the flow rate was 4mL/min, the culture temperature was 25°C, and the culture time was 10h; finally, yeast milk was obtained, and the dry yeast milk was measured by drying and weighing method.
  • the substance content is 18%.
  • the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion of 8% mass concentration;
  • the obtained thermally extracted product is centrifuged, wherein the centrifugation speed is 5000 rpm, and the centrifugation time is 20 minutes to obtain a supernatant, and the dry matter content of the supernatant is 3% by drying weighing method.
  • step (3) Enzymatic hydrolysis: 40 g of the supernatant obtained in step (2) was added to the enzymatic hydrolysis reaction tank, 0.006 g of vitamin C and 0.0012 g of papain were added, and the enzymatic hydrolysis was carried out at 40°C and pH 4.0 for 10 hours; then 0.0012 g of g flavor protease, hydrolyzed at 40°C and pH 4.0 for 5h.
  • step (3) The yeast extract solution after enzymolysis in step (3) is heated to 80° C. to kill the enzyme for 2 hours, and evaporated and concentrated to obtain a concentrated product, and the solid content of the concentrated product is 35%.
  • the yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast paste, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by feeding feed flow.
  • the culture time was 5h; then starvation culture was carried out, and nutrient solution was added by feeding feeding method, the flow rate was 2mL/min, the culture temperature was 35°C, and the culture time was 5h; finally, yeast milk was obtained, and the dryness of yeast milk was measured by drying and weighing method.
  • the substance content is 18%.
  • the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion with a mass concentration of 15%;
  • the obtained thermally extracted product is centrifuged, wherein the centrifugation speed is 5000 rpm, and the centrifugation time is 20 minutes to obtain a supernatant, and the dry matter content of the supernatant is 4% measured by drying weighing method.
  • Enzymatic hydrolysis take 50 g of the supernatant obtained in step (2), add 0.04 g of vitamin C and 0.03 g of papain, and perform enzymatic hydrolysis at 80°C and pH 7.5 for 15 hours; then add 0.03 g of flavor protease, and at 80 °C, pH 7.5 under the conditions of enzymatic hydrolysis for 5h.
  • step (3) The yeast extract solution after enzymolysis in step (3) is heated to 80° C. to kill the enzyme for 2 hours, and evaporated and concentrated to obtain a concentrated product, and the solid content in the concentrated product is 38%.
  • the yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast paste, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by means of feed-feeding.
  • the culture time was 13h; then starvation culture was carried out, and nutrient solution was added by feeding feeding method, the flow rate was 5mL/min, the culture temperature was 30°C, and the culture time was 8h; finally, yeast milk was obtained, and the dried yeast milk was measured by drying and weighing method.
  • the substance content is 18%.
  • the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion with a mass concentration of 12%;
  • the obtained thermally extracted product is centrifuged, wherein the centrifugation speed is 5000 rpm, and the centrifugation time is 20 minutes to obtain a supernatant, and the dry matter content of the supernatant is 5.1% by drying and weighing.
  • Enzymatic hydrolysis take 45 g of the supernatant obtained in step (2), add 0.022 g of vitamin C and 0.022 g of papain, and enzymatically hydrolyze 13 hours at 60°C and pH 6.0; then add 0.022 g of flavor protease, and at 60 °C, pH 5.5 under the conditions of enzymatic hydrolysis for 8h.
  • step (3) The yeast extract solution after enzymolysis in step (3) is heated to 80° C. to kill the enzyme for 2 hours, and then evaporated and concentrated to obtain a concentrated product, and the solid content in the concentrated product is 40%.
  • the yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast paste, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by means of feed-feeding.
  • the culture time was 10h; then starvation culture was carried out, and the nutrient solution was added by the feeding method, the flow rate was 6mL/min, the culture temperature was 30°C, and the culture time was 5h; finally, yeast milk was obtained, and the dry yeast milk was measured by drying and weighing method.
  • the substance content is 18%.
  • the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion with a mass concentration of 12%;
  • the obtained thermally extracted product is centrifuged, wherein the centrifugation speed is 5000 rpm, and the centrifugation time is 20 minutes to obtain a supernatant, and the dry matter content of the supernatant is 4% measured by drying weighing method.
  • Enzymatic hydrolysis take 40 g of the supernatant obtained in step (2), add 0.016 g of VC, 0.008 g of alkaline protease, 0.008 g of trypsin and 0.008 g of neutral protease, and enzymatically hydrolyze 13 hours at 65°C and pH 6.5 ; Then add 0.008g flavor protease and 0.008g aminopeptidase, and enzymolysis for 8h at 60°C and pH 5.5.
  • step (3) The yeast extract solution after enzymolysis in step (3) is heated to 80° C. to kill the enzyme for 2 hours, and is evaporated and concentrated to obtain a concentrated product, and the solid content in the concentrated product is 42%.
  • the yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast paste, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by feeding feed flow.
  • the cultivation time was 15h; then starvation cultivation was carried out, and nutrient solution was added by feeding feeding method, the flow rate was 7mL/min, the cultivation temperature was 25°C, and the cultivation time was 9h; finally, yeast milk was obtained, and the dried yeast milk was measured by drying and weighing method.
  • the substance content is 18%.
  • the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion with a mass concentration of 10%;
  • the obtained thermally extracted product is centrifuged, wherein the centrifugation speed is 5000 rpm and the centrifugation time is 20 minutes to obtain a supernatant, and the dry matter content of the supernatant is 3.4% measured by drying weighing method.
  • Enzymatic hydrolysis take 40 g of the supernatant obtained in step (2), add 0.02 g of VC and 0.017 g of neutral protease, and perform enzymatic hydrolysis for 12 h at 70° C. and pH 5; then add 0.017 g of carboxypeptidase, and at 50 °C, pH 6 conditions for enzymatic hydrolysis for 7h.
  • step (3) The yeast extract solution after enzymolysis in step (3) is heated to 80° C. to kill the enzyme for 2 hours, and then evaporated and concentrated to obtain a concentrated product, and the solid content of the concentrated product is 45%.
  • the yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast paste, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by feeding feed flow.
  • the culture time was 8h; then starvation culture was carried out, and the nutrient solution was added by feeding feeding method, the flow rate was 8mL/min, the culture temperature was 25°C, and the culture time was 6h; finally, yeast milk was obtained, and the dryness of yeast milk was measured by drying and weighing method.
  • the substance content is 18%.
  • the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion with a mass concentration of 10%;
  • the obtained thermally extracted product was subjected to centrifugation, wherein the centrifugation rate was 5000 rpm and the centrifugation time was 20 minutes to obtain a supernatant, and the dry matter content of the supernatant was 3.3% measured by drying weighing method.
  • Enzymatic hydrolysis take 40 g of the supernatant obtained in step (2), add 0.017 g of VC and 0.007 g of alkaline protease, and perform enzymatic hydrolysis at 55°C and pH 4.5 for 11 h; then add 0.007 g of aminopeptidase, and at 70 The enzymatic hydrolysis was carried out at °C and pH 5 for 10 h.
  • step (3) The yeast extract solution after enzymolysis in step (3) is heated to 80° C. to kill the enzyme for 2 hours, and is evaporated and concentrated to obtain a concentrated product, and the solid content in the concentrated product is 42%.
  • the yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast extract, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by means of feed-feeding.
  • the culture time was 13h; then starvation culture was carried out, and nutrient solution was added by feeding feeding method, the flow rate was 5mL/min, the culture temperature was 30°C, and the culture time was 8h; finally, yeast milk was obtained, and the dried yeast milk was measured by drying and weighing method.
  • the substance content is 18%.
  • step (2) Enzymatic hydrolysis: the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion with a mass concentration of 12%, 45 g of the above-prepared yeast emulsion is added to the enzymatic hydrolysis reaction tank, and 0.054 g of vitamin C and 0.054 g of vitamin C are added. Papain was enzymatically hydrolyzed at 60°C and pH 6.0 for 13 hours; then, 0.054 g of flavor protease was added, and the enzyme was hydrolyzed at 60°C and pH 5.5 for 8 hours.
  • step (3) spray-drying the product after enzymolysis in step (2) to obtain a cysteine-rich yeast extract, wherein the spray-drying inlet air temperature is controlled at 155-175°C, and the outlet air temperature is controlled at 90- 115°C, the atomization pressure is controlled at 100-160bar, and the temperature of the material is controlled at 25-45°C.
  • the yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast paste, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by feeding feed flow, the nutrient solution flow rate is 20mL/min, the ventilation rate is 20L/min, and the culture temperature is 50°C.
  • the culture time was 22h; then starvation culture was carried out, and nutrient solution was added by feeding feeding method, the flow rate was 10mL/min, the culture temperature was 50°C, and the culture time was 3h; finally, yeast milk was obtained, and the dry yeast milk was measured by drying and weighing method.
  • the substance content is 20%.
  • the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion with a mass concentration of 12%;
  • the obtained thermally extracted product is subjected to centrifugation, wherein the centrifugation speed is 5000 rpm and the centrifugation time is 20 minutes to obtain a supernatant, and the dry matter content of the supernatant is 2% measured by drying weighing method.
  • Enzymatic hydrolysis take 45 g of the supernatant obtained in step (2), add 0.009 g of vitamin C and 0.009 g of papain, and perform enzymatic hydrolysis at 60°C and pH 6.0 for 13 hours; then add 0.009 g of flavor protease, and at 60 °C, pH 5.5 under the conditions of enzymatic hydrolysis for 8h.
  • step (3) (4) spray-drying the product after enzymolysis in step (3) to obtain a cysteine-rich yeast extract, wherein the spray-drying inlet air temperature is controlled at 155-175°C, and the outlet air temperature is controlled at 90- 115°C, the atomization pressure is controlled at 100-160bar, and the temperature of the material is controlled at 25-45°C.
  • the yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast paste, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by means of feed-feeding.
  • the culture time was 13h; then starvation culture was carried out, and nutrient solution was added by feeding feeding method, the flow rate was 5mL/min, the culture temperature was 30°C, and the culture time was 8h; finally, yeast milk was obtained, and the dried yeast milk was measured by drying and weighing method.
  • the substance content is 18%.
  • the obtained thermally extracted product is subjected to centrifugation, wherein the centrifugation speed is 5000 rpm and the centrifugation time is 20 minutes to obtain a supernatant, and the dry matter content of the supernatant is 2% measured by drying weighing method.
  • Enzymatic hydrolysis take 45 g of the supernatant obtained in step (2), add 0.009 g of vitamin C and 0.009 g of papain, and perform enzymatic hydrolysis at 60°C and pH 6.0 for 13 hours; then add 0.009 g of flavor protease, and at 60 °C, pH 5.5 under the conditions of enzymatic hydrolysis for 8h.
  • step (3) (4) spray-drying the product after enzymolysis in step (3) to obtain a cysteine-rich yeast extract, wherein the spray-drying inlet air temperature is controlled at 155-175°C, and the outlet air temperature is controlled at 90- 115°C, the atomization pressure is controlled at 100-160bar, and the temperature of the receiving material is controlled at 25-45°C.
  • the yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300 g of yeast paste, 600 g of peptone, 600 g of glucose and 28500 g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by means of feed-feeding.
  • the culture time was 13h; then starvation culture was carried out, and nutrient solution was added by feeding feeding method, the flow rate was 5mL/min, the culture temperature was 30°C, and the culture time was 8h; finally, yeast milk was obtained, and the dried yeast milk was measured by drying and weighing method.
  • the substance content is 18%.
  • the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion with a mass concentration of 12%;
  • the obtained thermally extracted product is subjected to centrifugation, wherein the centrifugation speed is 5000 rpm and the centrifugation time is 20 minutes to obtain a supernatant, and the dry matter content of the supernatant is 2% measured by drying weighing method.
  • step (3) Enzymatic hydrolysis: take 45 g of the supernatant obtained in step (2), add 0.027 g of vitamin C and 0.018 g of papain, and enzymatically hydrolyze 8 h at 30°C and pH 8.0; then add 0.018 g of flavor protease, and at 30 The enzymatic hydrolysis was carried out under the conditions of °C and pH 8 for 12 h.
  • step (3) (4) spray-drying the product after enzymolysis in step (3) to obtain a cysteine-rich yeast extract, wherein the spray-drying inlet air temperature is controlled at 155-175°C, and the outlet air temperature is controlled at 90- 115°C, the atomization pressure is controlled at 100-160bar, and the temperature of the material is controlled at 25-45°C.
  • the cysteine content in the yeast extract is tested by high performance liquid chromatography, and the detection method is as follows:
  • Chromatographic column Agilent SB-C18 column.
  • phase A is 0.1% phosphoric acid
  • phase B is ethanol, in a ratio of 80:20, and the gradient is equilibrated for 3-4 hours.
  • the flow rate is 0.6mL/min.
  • the standard series and the sample solution were injected separately, and the injection volume was 0.6uL.
  • the chromatographic peak of cysteine was qualitatively determined according to the retention time of the standard and the standard curve (the relationship curve between the concentration of the standard and the peak area) was drawn. Peak area, and the percentage of cysteine was calculated by the external standard method.
  • X the content of cysteine in the sample, %
  • the cysteine-rich yeast extract preparation method of the present invention can obtain yeast extracts with a cysteine content greater than or equal to 1%.

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Abstract

A cysteine-rich yeast extract and a preparation method therefor. The preparation method comprises the following steps: (1) fermenting an original strain to obtain yeast milk; (2) carrying out thermal extraction and centrifugation on the yeast milk obtained in step (1) to obtain a supernatant; and (3) carrying out enzymatic hydrolysis on the supernatant of step (2) to obtain cysteine-rich yeast extract. The original strain in step (1) is saccharomyces cerevisiae rich in reduced glutathione, which was deposited at China Center for Type Culture Collection (Wuhan) with a deposit number of CCTCC M 205130 on 25 October 2005.

Description

一种富含半胱氨酸的酵母抽提物及其制备方法A kind of yeast extract rich in cysteine and preparation method thereof 技术领域technical field
本发明涉及酵母抽提物领域，具体涉及一种富含半胱氨酸的酵母抽提物及其制备方法。The invention relates to the field of yeast extracts, in particular to a cysteine-rich yeast extract and a preparation method thereof.
背景技术Background technique
酵母抽提物，是以食用酵母为原料，通过将酵母细胞内的蛋白质、核酸等降解后精制而成的具有食品属性的天然调味料，其含有蛋白质、多肽、氨基酸、核苷酸等多种有效成分，具有增鲜、增厚味、回味持久的特点，被广泛应用于调味品行业。Yeast extract is a natural seasoning with food properties, which is made of edible yeast as raw material and refined by degrading proteins and nucleic acids in yeast cells. It contains proteins, polypeptides, amino acids, nucleotides, etc. The active ingredient has the characteristics of freshening, thickening and lasting aftertaste, and is widely used in the condiment industry.
半胱氨酸在食品加工中的作用：提供肉味的前体物质。The role of cysteine in food processing: a precursor for providing meaty taste.
目前，现有技术中所制备的酵母抽提物中半胱氨酸含量较低，一般在1％以下。At present, the cysteine content in the yeast extract prepared in the prior art is relatively low, generally below 1%.
同时，现有技术公开了一种用于将胱氨酸转化为半胱氨酸和/或谷胱甘肽的方法，其包括使胱氨酸与微生物接触。还公开了包含至少1.8mg/g的半胱氨酸的酵母提取和包含至少1.3mg/g的半胱氨酸的酵母自溶物。半胱氨酸含量1.8mg/g(即1.8‰)，含量远低于1％。Meanwhile, the prior art discloses a method for converting cystine to cysteine and/or glutathione, comprising contacting cystine with microorganisms. Also disclosed are yeast extracts comprising at least 1.8 mg/g cysteine and yeast autolysates comprising at least 1.3 mg/g cysteine. The cysteine content is 1.8mg/g (ie, 1.8‰), which is much lower than 1%.
发明内容SUMMARY OF THE INVENTION
本发明所解决的技术问题是，现有技术中酵母自溶物或酵母抽提物的半胱氨酸含量较低，远远低于1％。The technical problem solved by the present invention is that the cysteine content of yeast autolysate or yeast extract in the prior art is relatively low, far below 1%.
本发明提供了一种半胱氨酸含量在1％以上的酵母抽提物及其制备方法。The invention provides a yeast extract with a cysteine content of more than 1% and a preparation method thereof.
具体来说，本发明为了解决上述技术问题，提供了如下技术方案：Specifically, in order to solve the above-mentioned technical problems, the present invention provides the following technical solutions:
本发明提供了一种富含半胱氨酸的酵母抽提物，所述酵母抽提物通过包含下述步骤制备得到：The present invention provides a cysteine-rich yeast extract prepared by comprising the following steps:
(1)将原始菌株发酵得到酵母乳；(1) the original strain is fermented to obtain yeast milk;
(2)将步骤(1)所得酵母乳热提取、离心得到上清液；(2) the yeast milk obtained in step (1) is thermally extracted and centrifuged to obtain a supernatant;
(3)将步骤(2)所得上清液酶解，得到富含半胱氨酸的酵母抽提物；(3) enzymolysis of the supernatant obtained in step (2) to obtain a cysteine-rich yeast extract;
其中，步骤(1)中所述原始菌株为富含还原型谷胱甘肽的酵母菌株(saccharomyces cerevisiae)，保藏编号为：CCTCC M 205130，保藏地址为：于2005年10月25日保藏于中国典型培养物保藏中心(武汉)。Wherein, the original strain described in the step (1) is a yeast strain (saccharomyces cerevisiae) rich in reduced glutathione, the deposit number is: CCTCC M 205130, and the deposit address is: on October 25, 2005, it was preserved in China Type Culture Collection (Wuhan).
优选地，以酵母抽提物干物质的质量计，酵母抽提物中半胱氨酸含量≥1％，优选为所述酵母抽提物中半胱氨酸含量≥2％。Preferably, the cysteine content in the yeast extract is greater than or equal to 1%, preferably the cysteine content in the yeast extract is greater than or equal to 2% based on the mass of the yeast extract dry matter.
优选地，步骤(1)中所述发酵过程以补料流加方式添加营养液；优选的，发酵过程包含普通培养过程和饥饿培养过程，按重量百份比计，饥饿培养过程营养液添加流量为普通培养过程添加流量的10-40％。Preferably, the fermentation process described in step (1) adds nutrient solution in a fed-feed mode; preferably, the fermentation process includes an ordinary culture process and a starvation culture process, and in percentage by weight, the nutrient solution added flow rate in the starvation culture process Add 10-40% of the flow rate for normal culture procedures.
优选地，所述普通培养的温度为25-35℃，优选普通培养的时间为5-20h；进一步优选的，所述饥饿培养的温度为25-35℃，优选饥饿培养的时间为5-10h。Preferably, the temperature of the normal culture is 25-35°C, and the time of the normal culture is preferably 5-20h; further preferably, the temperature of the starvation culture is 25-35°C, and the time of the starvation culture is preferably 5-10h .
优选地，步骤(2)热提取所用酵母乳的质量浓度为8-15％；优选热提取的温度为80-110℃，进一步优选的95℃；优选提取时pH值为5.0-8.0；优选热提取时间为60-120min，进一步优选为90min。Preferably, the mass concentration of the yeast milk used in the thermal extraction in step (2) is 8-15%; the temperature of the thermal extraction is preferably 80-110°C, more preferably 95°C; the pH value during extraction is preferably 5.0-8.0; The extraction time is 60-120min, more preferably 90min.
优选地，步骤(3)酶解时，依次采用内切蛋白酶和外切蛋白酶进行酶解，优选所述内切蛋白酶选自碱性蛋白酶、木瓜蛋白酶、胰蛋白酶和中性蛋白酶中的一种或两种以上；进一步优选所述外切蛋白酶选自风味蛋白酶、羧肽酶和氨肽酶中的一种或两种以上。Preferably, during the enzymatic hydrolysis in step (3), endoprotease and exoprotease are used for enzymolysis in sequence, preferably the endoprotease is selected from one of alkaline protease, papain, trypsin and neutral protease or two or more; further preferably, the exoprotease is selected from one or more of flavor protease, carboxypeptidase and aminopeptidase.
优选地，以上清液干物质的质量计，所述内切蛋白酶的添加量为0.1-1.5％，优选为1％。Preferably, the added amount of the endoprotease is 0.1-1.5%, preferably 1%, based on the mass of the dry matter of the supernatant.
优选地，内切蛋白酶酶解时，酶解温度为40-80℃，优选的，酶解pH值为4.0-7.5，酶解时间为10-15h。Preferably, during the enzymatic hydrolysis by endoprotease, the enzymatic hydrolysis temperature is 40-80° C., preferably, the enzymatic hydrolysis pH value is 4.0-7.5, and the enzymatic hydrolysis time is 10-15 h.
优选地，以上清液干物质的质量计，所述外切蛋白酶的添加量为0.1-1.5％，优选为1％。Preferably, the amount of the exoprotease added is 0.1-1.5%, preferably 1%, based on the mass of the dry matter of the supernatant.
优选地，外切蛋白酶酶解时，酶解温度为40-80℃；优选的，酶解pH值为4.0-7.5，酶解时间为5-10h。Preferably, during the enzymatic hydrolysis by exoprotease, the enzymatic hydrolysis temperature is 40-80° C.; preferably, the enzymatic hydrolysis pH value is 4.0-7.5, and the enzymatic hydrolysis time is 5-10 h.
优选地，步骤(3)酶解前还包括：向步骤(2)得到的上清液中加入维生素C；优选的，以上清液干物质的质量计，所述维生素C的添加量为0.5-2％；进一步优选为1％。Preferably, before the enzymatic hydrolysis in step (3), it further comprises: adding vitamin C to the supernatant obtained in step (2); 2%; more preferably 1%.
优选地，步骤(3)后还包括下述步骤：步骤(3)后还包括下述步骤：将步骤(3)酶解后酶解液依次进行蒸发浓缩和干燥，得到富含半胱氨酸的酵母抽提物。Preferably, after step (3), it also includes the following steps: after step (3), it also includes the following steps: the enzymatic hydrolysis solution after the enzymatic hydrolysis in step (3) is successively evaporated, concentrated and dried to obtain cysteine-rich of yeast extract.
优选地，所述干燥方式为喷雾干燥、真空干燥或滚筒干燥中的一种,进一步优选为喷雾干燥，更进一步优选喷雾干燥进风温度控制在155-175℃，出风温度控制在90-115℃，雾化压力控制在100-160bar，收料温度控制在25-45℃。Preferably, the drying method is one of spray drying, vacuum drying or drum drying, more preferably spray drying, and even more preferably, the inlet air temperature of spray drying is controlled at 155-175°C, and the outlet air temperature is controlled at 90-115°C ℃, the atomization pressure is controlled at 100-160bar, and the receiving temperature is controlled at 25-45℃.
另一方面，本发明提供了一种富含半胱氨酸的酵母抽提物的制备方法，具体包括下述步骤：On the other hand, the invention provides a preparation method of a cysteine-rich yeast extract, which specifically comprises the following steps:
(1)将原始菌株发酵得到酵母乳；(1) the original strain is fermented to obtain yeast milk;
(2)将步骤(1)所得酵母乳热提取、离心得到上清液；(2) the yeast milk obtained in step (1) is thermally extracted and centrifuged to obtain a supernatant;
(3)将步骤(2)所得上清液酶解，得到富含半胱氨酸的酵母抽提物。(3) Enzymatic hydrolysis of the supernatant obtained in step (2) to obtain a cysteine-rich yeast extract.
本发明有益效果包括：The beneficial effects of the present invention include:
(1)本发明所用原料均为无毒无害的可食用原料，所制备的产品及副产品均可用于食品工业；(1) the raw materials used in the present invention are non-toxic and harmless edible raw materials, and the prepared products and by-products can be used in the food industry;
(2)本发明所提供的制备方法提取率高，产物酵母抽提物的半胱氨酸含量高，最高可达到2％。(2) The preparation method provided by the present invention has a high extraction rate, and the cysteine content of the product yeast extract is high, up to 2%.
具体实施方式Detailed ways
半胱氨酸存在于酵母抽提物中，在食品加工中可以用作提供肉味的前体物质，但现有技术所得的酵母抽提物中半胱氨酸含量较低。本发明人经过锐意研究后，提供了一种富含半胱氨酸的酵母抽提物及其制备方法。Cysteine exists in yeast extract and can be used as a precursor substance to provide meat taste in food processing, but the content of cysteine in yeast extract obtained in the prior art is low. After earnest research, the present inventor provides a cysteine-rich yeast extract and a preparation method thereof.
一方面，本发明提供了一种富含半胱氨酸的酵母抽提物，所述酵母抽提物通过包含下述步骤的制备方法制备得到：In one aspect, the present invention provides a cysteine-rich yeast extract prepared by a preparation method comprising the following steps:
(1)将原始菌株发酵得到酵母乳；(1) the original strain is fermented to obtain yeast milk;
(2)将步骤(1)所得酵母乳热提取、离心得到上清液；(2) the yeast milk obtained in step (1) is thermally extracted and centrifuged to obtain a supernatant;
(3)将步骤(2)所得上清液酶解，得到富含半胱氨酸的酵母抽提物；(3) enzymolysis of the supernatant obtained in step (2) to obtain a cysteine-rich yeast extract;
其中，步骤(1)中所述原始菌株为富含还原型谷胱甘肽的酵母菌株(saccharomyces cerevisiae)，保藏编号为：CCTCC M 205130，保藏地址为：于2005年10月25日保藏于中国典型培养物保藏中心(武汉)。Wherein, the original strain described in the step (1) is a yeast strain (saccharomyces cerevisiae) rich in reduced glutathione, the deposit number is: CCTCC M 205130, and the deposit address is: on October 25, 2005, it was preserved in China Type Culture Collection (Wuhan).
在本发明的一个优选实施方案中，以酵母抽提物干物质的质量计，酵母抽提物中半胱氨酸含量≥1％，优选为所述酵母抽提物中半胱氨酸含量≥2％。In a preferred embodiment of the present invention, the cysteine content in the yeast extract is greater than or equal to 1% based on the mass of the dry matter of the yeast extract, preferably the cysteine content in the yeast extract is greater than or equal to 1%. 2%.
在本发明的一个优选实施方案中，步骤(1)中所述发酵过程以补料流加方式添加营养液；优选的，发酵过程包含普通培养过程和饥饿培养过程，按重量百份比计，饥饿培养过程营养液添加流量为普通培养过程添加流量的10-40％，进一步优选的，所述营养液包括如下重量百分比，酵母膏1％、蛋白胨2％和葡萄糖2％。In a preferred embodiment of the present invention, the fermentation process described in step (1) adds nutrient solution in a fed-feed way; The addition flow of the nutrient solution in the starvation culture process is 10-40% of that in the ordinary culture process. Further preferably, the nutrient solution includes the following weight percentages: 1% of yeast extract, 2% of peptone and 2% of glucose.
在本发明的一个优选实施方案中，所述普通培养的温度为25-35℃，优选普通培养的时间为5-20h；进一步优选的，所述饥饿培养的温度为25-35℃，优选饥饿培养的时间为5-10h。In a preferred embodiment of the present invention, the temperature of the ordinary culture is 25-35°C, and preferably the time of the ordinary culture is 5-20h; further preferably, the temperature of the starvation culture is 25-35°C, preferably starvation The incubation time is 5-10h.
在本发明的一个优选实施方案中，步骤(2)热提取所用酵母乳的质量浓度为8-15％；优选热提取的温度为80-110℃，进一步优选的95℃；优选提取时pH值为5.0-8.0；优选热提取时间为60-120min，进一步优选为90min；在热抽提后将热提取产物进行离心处理去除杂质，得到上清液，其中离心速率为5000rpm，离心时间为20分钟。In a preferred embodiment of the present invention, the mass concentration of the yeast milk used in the thermal extraction in step (2) is 8-15%; the temperature of the thermal extraction is preferably 80-110°C, more preferably 95°C; the pH value during extraction is preferably is 5.0-8.0; the preferred thermal extraction time is 60-120min, more preferably 90min; after the thermal extraction, the thermally extracted product is subjected to centrifugal treatment to remove impurities to obtain a supernatant, wherein the centrifugal speed is 5000rpm, and the centrifugal time is 20 minutes .
在本发明的一个优选实施方案中，步骤(3)酶解时，依次采用内切蛋白酶和外切蛋白酶进行酶解，优选所述内切蛋白酶选自碱性蛋白酶、木瓜蛋白酶、胰蛋白酶和中性蛋白酶中的一种或两种以上；进一步优选所述外切蛋白酶选自风味蛋白酶、羧肽酶和氨肽酶中的一种或两种以上。In a preferred embodiment of the present invention, during the enzymatic hydrolysis in step (3), endoprotease and exoprotease are used for enzymolysis in sequence, preferably the endoprotease is selected from the group consisting of alkaline protease, papain, trypsin and intermediate protease. one or two or more of the protease; further preferably, the exoprotease is selected from one or more of flavor protease, carboxypeptidase and aminopeptidase.
在本发明的一个优选实施方案中，以上清液干物质的质量计，所述内切蛋白酶的添加量为0.1-1.5％，优选为1％。In a preferred embodiment of the present invention, the added amount of the endoprotease is 0.1-1.5%, preferably 1%, based on the mass of the supernatant dry matter.
在本发明的一个优选实施方案中，内切蛋白酶酶解时，酶解温度为40-80℃，优选的，酶解pH值为4.0-7.5，酶解时间为10-15h。In a preferred embodiment of the present invention, during the enzymatic hydrolysis by endoprotease, the enzymatic hydrolysis temperature is 40-80° C., preferably, the enzymatic hydrolysis pH value is 4.0-7.5, and the enzymatic hydrolysis time is 10-15 h.
在本发明的一个优选实施方案中，以上清液干物质的质量计，所述外切蛋 白酶的添加量为0.1-1.5％，优选为1％。In a preferred embodiment of the present invention, the amount of the exoprotease added is 0.1-1.5%, preferably 1%, based on the mass of the supernatant dry matter.
在本发明的一个优选实施方案中，外切蛋白酶酶解时，酶解温度为40-80℃；优选的，酶解pH值为4.0-7.5，酶解时间为5-10h。In a preferred embodiment of the present invention, during the enzymatic hydrolysis by the exoprotease, the enzymatic hydrolysis temperature is 40-80° C.; preferably, the enzymatic hydrolysis pH value is 4.0-7.5, and the enzymatic hydrolysis time is 5-10 h.
在本发明的一个优选实施方案中，步骤(3)酶解前还包括：向步骤(2)得到的上清液中加入维生素C；优选的，以上清液干物质的质量计，所述维生素C的添加量为0.5-2％；进一步优选为1％。In a preferred embodiment of the present invention, before step (3) enzymatic hydrolysis further comprises: adding vitamin C to the supernatant obtained in step (2); The addition amount of C is 0.5-2%; more preferably, it is 1%.
在本发明的一个优选实施方案中，步骤(3)后还包括下述步骤：将步骤(3)酶解后的酵母抽提物溶液升温至80℃灭酶2h，进行蒸发浓缩得到浓缩品，优选浓缩品固体含量在35-45％；将浓缩品进行干燥，得到富含半胱氨酸的酵母抽提物。优选地所述干燥方式为喷雾干燥、真空干燥或滚筒干燥中的一种,进一步优选为喷雾干燥，更进一步优选喷雾干燥进风温度控制在155-175℃，出风温度控制在90-115℃，雾化压力控制在100-160bar，收料温度控制在25-45℃。In a preferred embodiment of the present invention, after step (3), the following steps are further included: the yeast extract solution after enzymatic hydrolysis in step (3) is heated to 80° C. for 2 hours to inactivate enzymes, and evaporated and concentrated to obtain a concentrated product, Preferably, the solid content of the concentrate is 35-45%; the concentrate is dried to obtain a cysteine-rich yeast extract. Preferably, the drying method is one of spray drying, vacuum drying or drum drying, more preferably spray drying, and even more preferably, the inlet air temperature of spray drying is controlled at 155-175°C, and the outlet air temperature is controlled at 90-115°C , the atomization pressure is controlled at 100-160bar, and the receiving temperature is controlled at 25-45 ℃.
另一方面，本发明提供了一种富含半胱氨酸的酵母抽提物的制备方法，具体包括下述步骤：On the other hand, the invention provides a preparation method of a cysteine-rich yeast extract, which specifically comprises the following steps:
(1)将原始菌株发酵得到酵母乳；(1) the original strain is fermented to obtain yeast milk;
(2)将步骤(1)所得酵母乳热提取、离心得到上清液；(2) the yeast milk obtained in step (1) is thermally extracted and centrifuged to obtain a supernatant;
(3)将步骤(2)所得上清液酶解，得到富含半胱氨酸的酵母抽提物。(3) Enzymatic hydrolysis of the supernatant obtained in step (2) to obtain a cysteine-rich yeast extract.
下面将通过具体实施例进一步来说明本发明的富含半胱氨酸的酵母抽提物及其制备方法。The cysteine-rich yeast extract and the preparation method thereof of the present invention will be further described below through specific examples.
菌株保藏信息strain preservation information
本发明采用富含还原型谷胱甘肽的酵母菌株((saccharomyces cerevisiae)，保藏编号为：CCTCC M 205130，保藏地址为：于2005年10月25日保藏于中国典型培养物保藏中心(武汉)。该菌株在公开号为CN101575578A的专利公开文本中已有记载。The present invention adopts the yeast strain rich in reduced glutathione ((saccharomyces cerevisiae), the deposit number is: CCTCC M 205130, and the deposit address is: on October 25, 2005, it was deposited in the China Center for Type Culture Collection (Wuhan) The strain has been recorded in the patent publication with publication number CN101575578A.
本发明下述实施例中所用到各试剂和仪器来源如下：The sources of each reagent and instrument used in the following examples of the present invention are as follows:
表1 本发明实施例所用试剂和仪器Table 1 Reagents and instruments used in the examples of the present invention
试剂/仪器Reagents/Instruments 型号/纯度Model/Purity 生产和销售厂家Production and sales manufacturers
酵母膏Yeast extract BR500g生物试剂BR500g biological reagent 北京奥博星生物技术有限责任公司Beijing Aoboxing Biotechnology Co., Ltd.
蛋白胨Peptone 250g BR生化试剂250g BR biochemical reagent 北京奥博星生物技术有限责任公司Beijing Aoboxing Biotechnology Co., Ltd.
葡萄糖glucose 国药试剂D(+)-葡萄糖Sinopharm Reagent D(+)-glucose 国药集团化学试剂有限公司Sinopharm Group Chemical Reagent Co., Ltd.
碱性蛋白酶Alkaline protease 45万U/g450,000 U/g 夏盛实业集团Xia Sheng Industrial Group
木瓜蛋白酶papain 60万U/g600,000 U/g 南宁东恒华道生物科技有限责任公司Nanning Dongheng Huadao Biotechnology Co., Ltd.
胰蛋白酶trypsin 4000U/g4000U/g 南宁庞博生物工程有限公司Nanning Pangbo Biological Engineering Co., Ltd.
中性蛋白酶Neutral protease 5万U/g50,000 U/g 北京奥博星生物技术有限责任公司Beijing Aoboxing Biotechnology Co., Ltd.
风味蛋白酶flavor protease 1000U/g1000U/g 丹麦诺维信公司Novozymes, Denmark
羧肽酶carboxypeptidase 180U/mg180U/mg SolarbioSolarbio
氨肽酶aminopeptidase 3万U/g30,000 U/g 南宁东恒华道生物科技有限责任公司Nanning Dongheng Huadao Biotechnology Co., Ltd.
维生素CVitamin C 分析纯ARAnalytical pure AR 天津市鼎盛鑫化工有限公司Tianjin Dingshengxin Chemical Co., Ltd.
离心机centrifuge 3-15型Type 3-15 Sigma公司Sigma
喷雾干燥机spray dryer 1800L型1800L type 上海雅程仪器设备有限公司Shanghai Yacheng Instrument Equipment Co., Ltd.
液相色谱仪Liquid chromatography 1200型Model 1200 安捷伦公司Agilent
下面将通过实施例对本发明作进一步说明。The present invention will be further illustrated by the following examples.
下面实施例和对比例中酵母乳干物质含量和上清液干物质含量采用下述烘干称重法测定：In the following examples and comparative examples, the dry matter content of yeast milk and the dry matter content of the supernatant are measured by the following drying weighing method:
(1)将洗净的蒸发皿放入烘箱内，在105±2℃烘1h后，放入干燥器内，冷却称量，重复将蒸发皿烘干、称量，直至恒重；(1) Put the washed evaporating dish into an oven, dry it at 105±2°C for 1 h, put it into a desiccator, cool and weigh, repeat drying and weighing the evaporating dish until it has a constant weight;
(2)吸取10g样品放入已恒重的蒸发皿中，将蒸发皿放入烘箱内，在105±2℃烘1h后，放入干燥器内，冷却称量，重复将蒸发皿烘干、称量，直至恒重；(2) Take 10g of sample and put it into an evaporating dish with constant weight, put the evaporating dish in an oven, dry it at 105±2°C for 1 h, put it in a desiccator, cool and weigh, repeat drying the evaporating dish, Weigh until constant weight;
(3)按下式计算干物质含量：(3) Calculate the dry matter content as follows:
(W2-W1)/m*100％(W2-W1)/m*100%
其中：W1——蒸发皿重量，gAmong them: W1 - the weight of the evaporating dish, g
W2——蒸发皿和干物质的重量，gW2 - weight of evaporating dish and dry matter, g
m——样品的重量，gm - the weight of the sample, g
实施例1Example 1
(1)酵母发酵培养(1) Yeast fermentation culture
酵母菌种是富含还原型谷胱甘肽酵母菌株(saccharomyces cerevisiae)，保藏编号为CCTCC M 205130，向配料罐中加入酵母膏300g、蛋白胨600g、葡萄糖600和28500g纯化水配制成营养液备用，向发酵罐中加入100g营养液作为打底，接种酵母菌种进行普通培养，采用补料流加方式添加营养液，营养液流量为20mL/min，通风量为10L/min，培养温度25℃，培养时间20h；然后进行饥饿培养，采用补料流加方式添加营养液，流量为4mL/min，培养温度25℃，培养时间10h；最终得到酵母乳，采用烘干称重法测得酵母乳干物质含量为18％。The yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast paste, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by feeding feed flow. The culture time was 20h; then starvation culture was carried out, and the nutrient solution was added by the feeding method, the flow rate was 4mL/min, the culture temperature was 25°C, and the culture time was 10h; finally, yeast milk was obtained, and the dry yeast milk was measured by drying and weighing method. The substance content is 18%.
(2)热提取：(2) Thermal extraction:
首先将步骤(1)所得酵母乳与纯化水配制成8％质量浓度的酵母乳液；First, the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion of 8% mass concentration;
向提取罐中加入上述酵母乳液100g进行热提取，热提取的温度为80℃，热提取时溶液的pH值为5.0，热提取时间为60分钟，Add 100 g of the above yeast emulsion to the extraction tank for thermal extraction, the temperature of thermal extraction is 80°C, the pH value of the solution during thermal extraction is 5.0, and the thermal extraction time is 60 minutes,
并将所得热提取产物进行离心处理，其中，离心速率为5000rpm，离心时间为20分钟，得到上清液，采用烘干称重法测得上清液干物质含量为3％。The obtained thermally extracted product is centrifuged, wherein the centrifugation speed is 5000 rpm, and the centrifugation time is 20 minutes to obtain a supernatant, and the dry matter content of the supernatant is 3% by drying weighing method.
(3)酶解：向酶解反应罐中加入步骤(2)所得上清液40g，添加0.006g维生素C和0.0012g木瓜蛋白酶，在40℃、pH值4.0条件下酶解10h；之后添加0.0012g风味蛋白酶，在40℃、pH值4.0的条件下酶解5h。(3) Enzymatic hydrolysis: 40 g of the supernatant obtained in step (2) was added to the enzymatic hydrolysis reaction tank, 0.006 g of vitamin C and 0.0012 g of papain were added, and the enzymatic hydrolysis was carried out at 40°C and pH 4.0 for 10 hours; then 0.0012 g of g flavor protease, hydrolyzed at 40°C and pH 4.0 for 5h.
(4)将步骤(3)酶解后的酵母抽提物溶液升温至80℃灭酶2h，进行蒸发浓缩得到浓缩品，浓缩品中的固形物含量为35％。(4) The yeast extract solution after enzymolysis in step (3) is heated to 80° C. to kill the enzyme for 2 hours, and evaporated and concentrated to obtain a concentrated product, and the solid content of the concentrated product is 35%.
(5)将浓缩品进行喷雾干燥，得到富含半胱氨酸的酵母抽提物，其中喷雾干燥进风温度控制在155-175℃，出风温度控制在90-115℃，雾化压力控制在100-160bar，收料温度控制在25-45℃。(5) spray-drying the concentrated product to obtain a cysteine-rich yeast extract, wherein the spray-drying inlet air temperature is controlled at 155-175°C, the outlet air temperature is controlled at 90-115°C, and the atomization pressure is controlled At 100-160 bar, the receiving temperature is controlled at 25-45°C.
实施例2Example 2
(1)酵母发酵培养(1) Yeast fermentation culture
酵母菌种是富含还原型谷胱甘肽酵母菌株(saccharomyces cerevisiae)，保藏编号为CCTCC M 205130，向配料罐中加入酵母膏300g、蛋白胨600g、葡萄糖600和28500g纯化水配制成营养液备用，向发酵罐中加入100g营养液作为打底，接种酵母菌种进行普通培养，采用补料流加方式添加营养液，营养液流量为20mL/min，通风量为30L/min，培养温度35℃，培养时间5h；然后进行饥饿培养，采用补料流加方式添加营养液，流量为2mL/min，培养温度35℃，培养时间5h；最终得到酵母乳，采用烘干称重法测得酵母乳干物质含量为18％。The yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast paste, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by feeding feed flow. The culture time was 5h; then starvation culture was carried out, and nutrient solution was added by feeding feeding method, the flow rate was 2mL/min, the culture temperature was 35°C, and the culture time was 5h; finally, yeast milk was obtained, and the dryness of yeast milk was measured by drying and weighing method. The substance content is 18%.
(2)热提取：(2) Thermal extraction:
首先将步骤(1)所得酵母乳与纯化水配制成15％质量浓度的酵母乳液；First, the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion with a mass concentration of 15%;
向提取罐中加入上述酵母乳液100g进行热提取，热提取的温度为110℃，热提取时溶液的pH值为8.0，热提取时间为120分钟，Add 100 g of the above yeast emulsion to the extraction tank for thermal extraction, the temperature of thermal extraction is 110°C, the pH value of the solution during thermal extraction is 8.0, and the thermal extraction time is 120 minutes,
并将所得热提取产物进行离心处理，其中，离心速率为5000rpm，离心时间为20分钟，得到上清液，采用烘干称重法测得上清液干物质含量为4％。The obtained thermally extracted product is centrifuged, wherein the centrifugation speed is 5000 rpm, and the centrifugation time is 20 minutes to obtain a supernatant, and the dry matter content of the supernatant is 4% measured by drying weighing method.
(3)酶解：取步骤(2)所得上清液50g，添加0.04g维生素C和0.03g木瓜蛋白酶，在80℃、pH值7.5条件下酶解15h；之后添加0.03g风味蛋白酶，在80℃、pH值7.5的条件下酶解5h。(3) Enzymatic hydrolysis: take 50 g of the supernatant obtained in step (2), add 0.04 g of vitamin C and 0.03 g of papain, and perform enzymatic hydrolysis at 80°C and pH 7.5 for 15 hours; then add 0.03 g of flavor protease, and at 80 ℃, pH 7.5 under the conditions of enzymatic hydrolysis for 5h.
(4)将步骤(3)酶解后的酵母抽提物溶液升温至80℃灭酶2h，进行蒸发浓缩得到浓缩品，浓缩品中的固形物含量为38％。(4) The yeast extract solution after enzymolysis in step (3) is heated to 80° C. to kill the enzyme for 2 hours, and evaporated and concentrated to obtain a concentrated product, and the solid content in the concentrated product is 38%.
(5)将浓缩品进行喷雾干燥，得到富含半胱氨酸的酵母抽提物，其中喷雾干燥进风温度控制在155-175℃，出风温度控制在90-115℃，雾化压力控制在100-160bar，收料温度控制在25-45℃。(5) spray-drying the concentrated product to obtain a cysteine-rich yeast extract, wherein the spray-drying inlet air temperature is controlled at 155-175°C, the outlet air temperature is controlled at 90-115°C, and the atomization pressure is controlled At 100-160 bar, the receiving temperature is controlled at 25-45°C.
实施例3Example 3
(1)酵母发酵培养(1) Yeast fermentation culture
酵母菌种是富含还原型谷胱甘肽酵母菌株(saccharomyces cerevisiae)，保藏编号为CCTCC M 205130，向配料罐中加入酵母膏300g、蛋白胨600g、葡萄糖600和28500g纯化水配制成营养液备用，向发酵罐中加入100g营养液作 为打底，接种酵母菌种进行普通培养，采用补料流加方式添加营养液，营养液流量为20mL/min，通风量为20L/min，培养温度30℃，培养时间13h；然后进行饥饿培养，采用补料流加方式添加营养液，流量为5mL/min，培养温度30℃，培养时间8h；最终得到酵母乳，采用烘干称重法测得酵母乳干物质含量为18％。The yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast paste, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by means of feed-feeding. The culture time was 13h; then starvation culture was carried out, and nutrient solution was added by feeding feeding method, the flow rate was 5mL/min, the culture temperature was 30°C, and the culture time was 8h; finally, yeast milk was obtained, and the dried yeast milk was measured by drying and weighing method. The substance content is 18%.
(2)热提取：(2) Thermal extraction:
首先将步骤(1)所得酵母乳与纯化水配制成12％质量浓度的酵母乳液；First, the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion with a mass concentration of 12%;
向提取罐中加入上述酵母乳液100g进行热提取，热提取的温度为95℃，热提取时溶液的pH值为7.0，热提取时间为90分钟，Add 100 g of the above yeast emulsion to the extraction tank for thermal extraction, the temperature of thermal extraction is 95°C, the pH value of the solution during thermal extraction is 7.0, and the thermal extraction time is 90 minutes,
并将所得热提取产物进行离心处理，其中，离心速率为5000rpm，离心时间为20分钟，得到上清液，采用烘干称重法测得上清液干物质含量为5.1％。The obtained thermally extracted product is centrifuged, wherein the centrifugation speed is 5000 rpm, and the centrifugation time is 20 minutes to obtain a supernatant, and the dry matter content of the supernatant is 5.1% by drying and weighing.
(3)酶解：取步骤(2)所得上清液45g，添加0.022g维生素C和0.022g木瓜蛋白酶，在60℃、pH值6.0条件下酶解13h；之后添加0.022g风味蛋白酶，在60℃、pH值5.5的条件下酶解8h。(3) Enzymatic hydrolysis: take 45 g of the supernatant obtained in step (2), add 0.022 g of vitamin C and 0.022 g of papain, and enzymatically hydrolyze 13 hours at 60°C and pH 6.0; then add 0.022 g of flavor protease, and at 60 ℃, pH 5.5 under the conditions of enzymatic hydrolysis for 8h.
(4)将步骤(3)酶解后的酵母抽提物溶液升温至80℃灭酶2h，进行蒸发浓缩得到浓缩品，浓缩品中的固形物含量为40％。(4) The yeast extract solution after enzymolysis in step (3) is heated to 80° C. to kill the enzyme for 2 hours, and then evaporated and concentrated to obtain a concentrated product, and the solid content in the concentrated product is 40%.
(5)将浓缩品进行喷雾干燥，得到富含半胱氨酸的酵母抽提物，其中喷雾干燥进风温度控制在155-175℃，出风温度控制在90-115℃，雾化压力控制在100-160bar，收料温度控制在25-45℃。(5) spray-drying the concentrated product to obtain a cysteine-rich yeast extract, wherein the spray-drying inlet air temperature is controlled at 155-175°C, the outlet air temperature is controlled at 90-115°C, and the atomization pressure is controlled At 100-160 bar, the receiving temperature is controlled at 25-45°C.
实施例4Example 4
(1)酵母发酵培养(1) Yeast fermentation culture
酵母菌种是富含还原型谷胱甘肽酵母菌株(saccharomyces cerevisiae)，保藏编号为CCTCC M 205130，向配料罐中加入酵母膏300g、蛋白胨600g、葡萄糖600和28500g纯化水配制成营养液备用，向发酵罐中加入100g营养液作为打底，接种酵母菌种进行普通培养，采用补料流加方式添加营养液，营养液流量为20mL/min，通风量为20L/min，培养温度30℃，培养时间10h；然后进行饥饿培养，采用补料流加方式添加营养液，流量为6mL/min，培养温度30℃，培养时间5h；最终得到酵母乳，采用烘干称重法测得酵母乳干物质含量为18％。The yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast paste, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by means of feed-feeding. The culture time was 10h; then starvation culture was carried out, and the nutrient solution was added by the feeding method, the flow rate was 6mL/min, the culture temperature was 30°C, and the culture time was 5h; finally, yeast milk was obtained, and the dry yeast milk was measured by drying and weighing method. The substance content is 18%.
(2)热提取：(2) Thermal extraction:
首先将步骤(1)所得酵母乳与纯化水配制成12％质量浓度的酵母乳液；First, the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion with a mass concentration of 12%;
向提取罐中加入上述酵母乳液100g进行热提取，热提取的温度为95℃，热提取时溶液的pH值为7.0，热提取时间为90分钟，Add 100 g of the above yeast emulsion to the extraction tank for thermal extraction, the temperature of thermal extraction is 95°C, the pH value of the solution during thermal extraction is 7.0, and the thermal extraction time is 90 minutes,
并将所得热提取产物进行离心处理，其中，离心速率为5000rpm，离心时间为20分钟，得到上清液，采用烘干称重法测得上清液干物质含量为4％。The obtained thermally extracted product is centrifuged, wherein the centrifugation speed is 5000 rpm, and the centrifugation time is 20 minutes to obtain a supernatant, and the dry matter content of the supernatant is 4% measured by drying weighing method.
(3)酶解：取步骤(2)所得上清液40g，添加0.016gVC、0.008g碱性蛋白酶、0.008g胰蛋白酶和0.008g中性蛋白酶，在65℃、pH为6.5条件下酶解13h；之后添加0.008g风味蛋白酶和0.008g氨肽酶，在60℃、pH为5.5条件下酶解8h。(3) Enzymatic hydrolysis: take 40 g of the supernatant obtained in step (2), add 0.016 g of VC, 0.008 g of alkaline protease, 0.008 g of trypsin and 0.008 g of neutral protease, and enzymatically hydrolyze 13 hours at 65°C and pH 6.5 ; Then add 0.008g flavor protease and 0.008g aminopeptidase, and enzymolysis for 8h at 60°C and pH 5.5.
(4)将步骤(3)酶解后的酵母抽提物溶液升温至80℃灭酶2h，进行蒸发浓缩得到浓缩品，浓缩品中的固形物含量为42％。(4) The yeast extract solution after enzymolysis in step (3) is heated to 80° C. to kill the enzyme for 2 hours, and is evaporated and concentrated to obtain a concentrated product, and the solid content in the concentrated product is 42%.
(5)将浓缩品行喷雾干燥，得到富含半胱氨酸的酵母抽提物，其中喷雾干燥进风温度控制在155-175℃，出风温度控制在90-115℃，雾化压力控制在100-160bar，收料温度控制在25-45℃。(5) spray drying the concentrated product to obtain a cysteine-rich yeast extract, wherein the spray drying air inlet temperature is controlled at 155-175°C, the outlet air temperature is controlled at 90-115°C, and the atomization pressure is controlled at 100-160bar, the receiving temperature is controlled at 25-45℃.
实施例5Example 5
(1)酵母发酵培养(1) Yeast fermentation culture
酵母菌种是富含还原型谷胱甘肽酵母菌株(saccharomyces cerevisiae)，保藏编号为CCTCC M 205130，向配料罐中加入酵母膏300g、蛋白胨600g、葡萄糖600和28500g纯化水配制成营养液备用，向发酵罐中加入100g营养液作为打底，接种酵母菌种进行普通培养，采用补料流加方式添加营养液，营养液流量为20mL/min，通风量为20L/min，培养温度25℃，培养时间15h；然后进行饥饿培养，采用补料流加方式添加营养液，流量为7mL/min，培养温度25℃，培养时间9h；最终得到酵母乳，采用烘干称重法测得酵母乳干物质含量为18％。The yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast paste, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by feeding feed flow. The cultivation time was 15h; then starvation cultivation was carried out, and nutrient solution was added by feeding feeding method, the flow rate was 7mL/min, the cultivation temperature was 25°C, and the cultivation time was 9h; finally, yeast milk was obtained, and the dried yeast milk was measured by drying and weighing method. The substance content is 18%.
(2)热提取：(2) Thermal extraction:
首先将步骤(1)所得酵母乳与纯化水配制成10％质量浓度的酵母乳液；First, the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion with a mass concentration of 10%;
向提取罐中加入上述酵母乳液100g进行热提取，热提取的温度为90℃，热提取时溶液的pH值为6.0，热提取时间为80分钟，Add 100 g of the above yeast emulsion to the extraction tank for thermal extraction, the temperature of thermal extraction is 90°C, the pH value of the solution during thermal extraction is 6.0, and the thermal extraction time is 80 minutes,
并将所得热提取产物进行离心处理，其中，离心速率为5000rpm，离心时间为20分钟，得到上清液，采用烘干称重法测得上清液干物质含量为3.4％。The obtained thermally extracted product is centrifuged, wherein the centrifugation speed is 5000 rpm and the centrifugation time is 20 minutes to obtain a supernatant, and the dry matter content of the supernatant is 3.4% measured by drying weighing method.
(3)酶解：取步骤(2)所得上清液40g，添加0.02gVC、0.017g中性蛋白酶，在70℃、pH为5条件下酶解12h；之后添加0.017g羧肽酶，在50℃、pH为6条件下酶解7h。(3) Enzymatic hydrolysis: take 40 g of the supernatant obtained in step (2), add 0.02 g of VC and 0.017 g of neutral protease, and perform enzymatic hydrolysis for 12 h at 70° C. and pH 5; then add 0.017 g of carboxypeptidase, and at 50 ℃, pH 6 conditions for enzymatic hydrolysis for 7h.
(4)将步骤(3)酶解后的酵母抽提物溶液升温至80℃灭酶2h，进行蒸发浓缩得到浓缩品，浓缩品中的固形物含量为45％。(4) The yeast extract solution after enzymolysis in step (3) is heated to 80° C. to kill the enzyme for 2 hours, and then evaporated and concentrated to obtain a concentrated product, and the solid content of the concentrated product is 45%.
(5)将浓缩品进行喷雾干燥，得到富含半胱氨酸的酵母抽提物，其中喷雾干燥进风温度控制在155-175℃，出风温度控制在90-115℃，雾化压力控制在100-160bar，收料温度控制在25-45℃。(5) spray-drying the concentrated product to obtain a cysteine-rich yeast extract, wherein the spray-drying inlet air temperature is controlled at 155-175°C, the outlet air temperature is controlled at 90-115°C, and the atomization pressure is controlled At 100-160 bar, the receiving temperature is controlled at 25-45°C.
实施例6Example 6
(1)酵母发酵培养(1) Yeast fermentation culture
酵母菌种是富含还原型谷胱甘肽酵母菌株(saccharomyces cerevisiae)，保藏编号为CCTCC M 205130，向配料罐中加入酵母膏300g、蛋白胨600g、葡萄糖600和28500g纯化水配制成营养液备用，向发酵罐中加入100g营养液作为打底，接种酵母菌种进行普通培养，采用补料流加方式添加营养液，营养液流量为20mL/min，通风量为25L/min，培养温度30℃，培养时间8h；然后进行饥饿培养，采用补料流加方式添加营养液，流量为8mL/min，培养温度25℃，培养时间6h；最终得到酵母乳，采用烘干称重法测得酵母乳干物质含量为18％。The yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast paste, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by feeding feed flow. The culture time was 8h; then starvation culture was carried out, and the nutrient solution was added by feeding feeding method, the flow rate was 8mL/min, the culture temperature was 25°C, and the culture time was 6h; finally, yeast milk was obtained, and the dryness of yeast milk was measured by drying and weighing method. The substance content is 18%.
(2)热提取：(2) Thermal extraction:
首先将步骤(1)所得酵母乳与纯化水配制成10％质量浓度的酵母乳液；First, the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion with a mass concentration of 10%;
向提取罐中加入上述酵母乳液100g进行热提取，热提取的温度为105℃，热提取时溶液的pH值为5.0，热提取时间为70分钟，Add 100 g of the above yeast emulsion to the extraction tank for thermal extraction, the temperature of thermal extraction is 105°C, the pH value of the solution during thermal extraction is 5.0, and the thermal extraction time is 70 minutes,
并将所得热提取产物进行离心处理，其中，离心速率为5000rpm，离心时间为20分钟，得到上清液，采用烘干称重法测得上清液干物质含量为3.3％。The obtained thermally extracted product was subjected to centrifugation, wherein the centrifugation rate was 5000 rpm and the centrifugation time was 20 minutes to obtain a supernatant, and the dry matter content of the supernatant was 3.3% measured by drying weighing method.
(3)酶解：取步骤(2)所得上清液40g，添加0.017gVC、0.007g碱性蛋白酶，在55℃、pH为4.5条件下酶解11h；之后添加0.007g氨肽酶，在70℃、pH为5条件下酶解10h。(3) Enzymatic hydrolysis: take 40 g of the supernatant obtained in step (2), add 0.017 g of VC and 0.007 g of alkaline protease, and perform enzymatic hydrolysis at 55°C and pH 4.5 for 11 h; then add 0.007 g of aminopeptidase, and at 70 The enzymatic hydrolysis was carried out at ℃ and pH 5 for 10 h.
(4)将步骤(3)酶解后的酵母抽提物溶液升温至80℃灭酶2h，进行蒸发浓缩得到浓缩品，浓缩品中的固形物含量为42％。(4) The yeast extract solution after enzymolysis in step (3) is heated to 80° C. to kill the enzyme for 2 hours, and is evaporated and concentrated to obtain a concentrated product, and the solid content in the concentrated product is 42%.
(5)将浓缩品进行喷雾干燥，得到富含半胱氨酸的酵母抽提物，其中喷雾干燥进风温度控制在155-175℃，出风温度控制在90-115℃，雾化压力控制在100-160bar，收料温度控制在25-45℃。(5) spray-drying the concentrated product to obtain a cysteine-rich yeast extract, wherein the spray-drying inlet air temperature is controlled at 155-175°C, the outlet air temperature is controlled at 90-115°C, and the atomization pressure is controlled At 100-160 bar, the receiving temperature is controlled at 25-45°C.
对比例1Comparative Example 1
(1)酵母发酵培养(1) Yeast fermentation culture
酵母菌种是富含还原型谷胱甘肽酵母菌株(saccharomyces cerevisiae)，保藏编号为CCTCC M 205130，向配料罐中加入酵母膏300g、蛋白胨600g、葡萄糖600和28500g纯化水配制成营养液备用，向发酵罐中加入100g营养液作为打底，接种酵母菌种进行普通培养，采用补料流加方式添加营养液，营养液流量为20mL/min，通风量为20L/min，培养温度30℃，培养时间13h；然后进行饥饿培养，采用补料流加方式添加营养液，流量为5mL/min，培养温度30℃，培养时间8h；最终得到酵母乳，采用烘干称重法测得酵母乳干物质含量为18％。The yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast extract, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by means of feed-feeding. The culture time was 13h; then starvation culture was carried out, and nutrient solution was added by feeding feeding method, the flow rate was 5mL/min, the culture temperature was 30°C, and the culture time was 8h; finally, yeast milk was obtained, and the dried yeast milk was measured by drying and weighing method. The substance content is 18%.
(2)酶解：将步骤(1)所得酵母乳与纯化水配制成12％质量浓度的酵母乳液，向酶解反应罐中加入上述配制好的酵母乳液45g，添加0.054g维生素C和0.054g木瓜蛋白酶，在60℃、pH值6.0条件下酶解13h；之后添加0.054g风味蛋白酶，在60℃、pH值5.5的条件下酶解8h。(2) Enzymatic hydrolysis: the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion with a mass concentration of 12%, 45 g of the above-prepared yeast emulsion is added to the enzymatic hydrolysis reaction tank, and 0.054 g of vitamin C and 0.054 g of vitamin C are added. Papain was enzymatically hydrolyzed at 60°C and pH 6.0 for 13 hours; then, 0.054 g of flavor protease was added, and the enzyme was hydrolyzed at 60°C and pH 5.5 for 8 hours.
(3)将步骤(2)酶解后的产物进行喷雾干燥，得到富含半胱氨酸的酵母抽提物，其中喷雾干燥进风温度控制在155-175℃，出风温度控制在90-115℃，雾化压力控制在100-160bar，收料温度控制在25-45℃。(3) spray-drying the product after enzymolysis in step (2) to obtain a cysteine-rich yeast extract, wherein the spray-drying inlet air temperature is controlled at 155-175°C, and the outlet air temperature is controlled at 90- 115℃, the atomization pressure is controlled at 100-160bar, and the temperature of the material is controlled at 25-45℃.
对比例2Comparative Example 2
(1)酵母发酵培养(1) Yeast fermentation culture
酵母菌种是富含还原型谷胱甘肽酵母菌株(saccharomyces cerevisiae)，保藏编号为CCTCC M 205130，向配料罐中加入酵母膏300g、蛋白胨600g、葡萄糖600和28500g纯化水配制成营养液备用，向发酵罐中加入100g营养液作为打底，接种酵母菌种进行普通培养，采用补料流加方式添加营养液，营养液流量为20mL/min，通风量为20L/min，培养温度50℃，培养时间22h；然后进行饥饿培养，采用补料流加方式添加营养液，流量为10mL/min，培养温度50℃，培养时间3h；最终得到酵母乳，采用烘干称重法测得酵母乳干物质含量为20％。The yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast paste, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by feeding feed flow, the nutrient solution flow rate is 20mL/min, the ventilation rate is 20L/min, and the culture temperature is 50°C. The culture time was 22h; then starvation culture was carried out, and nutrient solution was added by feeding feeding method, the flow rate was 10mL/min, the culture temperature was 50°C, and the culture time was 3h; finally, yeast milk was obtained, and the dry yeast milk was measured by drying and weighing method. The substance content is 20%.
(2)热提取：(2) Thermal extraction:
首先将步骤(1)所得酵母乳与纯化水配制成12％质量浓度的酵母乳液；First, the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion with a mass concentration of 12%;
向提取罐中加入上述酵母乳液100g进行热提取，热提取的温度为95℃，热提取时溶液的pH值为7.0，热提取时间为90分钟，Add 100 g of the above yeast emulsion to the extraction tank for thermal extraction, the temperature of thermal extraction is 95°C, the pH value of the solution during thermal extraction is 7.0, and the thermal extraction time is 90 minutes,
并将所得热提取产物进行离心处理，其中，离心速率为5000rpm，离心时间为20分钟，得到上清液，采用烘干称重法测得上清液干物质含量为2％。The obtained thermally extracted product is subjected to centrifugation, wherein the centrifugation speed is 5000 rpm and the centrifugation time is 20 minutes to obtain a supernatant, and the dry matter content of the supernatant is 2% measured by drying weighing method.
(3)酶解：取步骤(2)所得上清液45g，添加0.009g维生素C和0.009g木瓜蛋白酶，在60℃、pH值6.0条件下酶解13h；之后添加0.009g风味蛋白酶，在60℃、pH值5.5的条件下酶解8h。(3) Enzymatic hydrolysis: take 45 g of the supernatant obtained in step (2), add 0.009 g of vitamin C and 0.009 g of papain, and perform enzymatic hydrolysis at 60°C and pH 6.0 for 13 hours; then add 0.009 g of flavor protease, and at 60 ℃, pH 5.5 under the conditions of enzymatic hydrolysis for 8h.
(4)将步骤(3)酶解后的产物进行喷雾干燥，得到富含半胱氨酸的酵母抽提物，其中喷雾干燥进风温度控制在155-175℃，出风温度控制在90-115℃，雾化压力控制在100-160bar，收料温度控制在25-45℃。(4) spray-drying the product after enzymolysis in step (3) to obtain a cysteine-rich yeast extract, wherein the spray-drying inlet air temperature is controlled at 155-175°C, and the outlet air temperature is controlled at 90- 115℃, the atomization pressure is controlled at 100-160bar, and the temperature of the material is controlled at 25-45℃.
对比例3Comparative Example 3
(1)酵母发酵培养(1) Yeast fermentation culture
酵母菌种是富含还原型谷胱甘肽酵母菌株(saccharomyces cerevisiae)，保藏编号为CCTCC M 205130，向配料罐中加入酵母膏300g、蛋白胨600g、葡萄糖600和28500g纯化水配制成营养液备用，向发酵罐中加入100g营养液作为打底，接种酵母菌种进行普通培养，采用补料流加方式添加营养液，营养液流量为20mL/min，通风量为20L/min，培养温度30℃，培养时间13h；然后进 行饥饿培养，采用补料流加方式添加营养液，流量为5mL/min，培养温度30℃，培养时间8h；最终得到酵母乳，采用烘干称重法测得酵母乳干物质含量为18％。The yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300g of yeast paste, 600g of peptone, 600g of glucose and 28,500g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by means of feed-feeding. The culture time was 13h; then starvation culture was carried out, and nutrient solution was added by feeding feeding method, the flow rate was 5mL/min, the culture temperature was 30°C, and the culture time was 8h; finally, yeast milk was obtained, and the dried yeast milk was measured by drying and weighing method. The substance content is 18%.
(2)热提取：(2) Thermal extraction:
向提取罐中加入上述酵母乳液100g进行热提取，热提取的温度为60℃，热提取时溶液的pH值为9，热提取时间为20分钟，Add 100 g of the above yeast emulsion to the extraction tank for thermal extraction, the temperature of thermal extraction is 60°C, the pH value of the solution during thermal extraction is 9, and the thermal extraction time is 20 minutes,
并将所得热提取产物进行离心处理，其中，离心速率为5000rpm，离心时间为20分钟，得到上清液，采用烘干称重法测得上清液干物质含量为2％。The obtained thermally extracted product is subjected to centrifugation, wherein the centrifugation speed is 5000 rpm and the centrifugation time is 20 minutes to obtain a supernatant, and the dry matter content of the supernatant is 2% measured by drying weighing method.
(3)酶解：取步骤(2)所得上清液45g，添加0.009g维生素C和0.009g木瓜蛋白酶，在60℃、pH值6.0条件下酶解13h；之后添加0.009g风味蛋白酶，在60℃、pH值5.5的条件下酶解8h。(3) Enzymatic hydrolysis: take 45 g of the supernatant obtained in step (2), add 0.009 g of vitamin C and 0.009 g of papain, and perform enzymatic hydrolysis at 60°C and pH 6.0 for 13 hours; then add 0.009 g of flavor protease, and at 60 ℃, pH 5.5 under the conditions of enzymatic hydrolysis for 8h.
(4)将步骤(3)酶解后的产物进行喷雾干燥，得到富含半胱氨酸的酵母抽提物，其中喷雾干燥进风温度控制在155-175℃，出风温度控制在90-115℃，雾化压力控制在100-160bar，收料温度控制在25-45℃。(4) spray-drying the product after enzymolysis in step (3) to obtain a cysteine-rich yeast extract, wherein the spray-drying inlet air temperature is controlled at 155-175°C, and the outlet air temperature is controlled at 90- 115°C, the atomization pressure is controlled at 100-160bar, and the temperature of the receiving material is controlled at 25-45°C.
对比例4Comparative Example 4
(1)酵母发酵培养(1) Yeast fermentation culture
酵母菌种是富含还原型谷胱甘肽酵母菌株(saccharomyces cerevisiae)，保藏编号为CCTCC M 205130，向配料罐中加入酵母膏300g、蛋白胨600g、葡萄糖600g和28500g纯化水配制成营养液备用，向发酵罐中加入100g营养液作为打底，接种酵母菌种进行普通培养，采用补料流加方式添加营养液，营养液流量为20mL/min，通风量为20L/min，培养温度30℃，培养时间13h；然后进行饥饿培养，采用补料流加方式添加营养液，流量为5mL/min，培养温度30℃，培养时间8h；最终得到酵母乳，采用烘干称重法测得酵母乳干物质含量为18％。The yeast strain is rich in reduced glutathione yeast strain (saccharomyces cerevisiae), the preservation number is CCTCC M 205130, and 300 g of yeast paste, 600 g of peptone, 600 g of glucose and 28500 g of purified water are added to the batching tank to prepare a nutrient solution for subsequent use, Add 100g of nutrient solution to the fermenter as a base, inoculate yeast strains for common culture, and add nutrient solution by means of feed-feeding. The culture time was 13h; then starvation culture was carried out, and nutrient solution was added by feeding feeding method, the flow rate was 5mL/min, the culture temperature was 30°C, and the culture time was 8h; finally, yeast milk was obtained, and the dried yeast milk was measured by drying and weighing method. The substance content is 18%.
(2)热提取：(2) Thermal extraction:
首先将步骤(1)所得酵母乳与纯化水配制成12％质量浓度的酵母乳液；First, the yeast milk obtained in step (1) and purified water are prepared into a yeast emulsion with a mass concentration of 12%;
向提取罐中加入上述酵母乳液100g进行热提取，热提取的温度为95℃，热提取时溶液的pH值为7.0，热提取时间为90分钟，Add 100 g of the above yeast emulsion to the extraction tank for thermal extraction, the temperature of thermal extraction is 95°C, the pH value of the solution during thermal extraction is 7.0, and the thermal extraction time is 90 minutes,
并将所得热提取产物进行离心处理，其中，离心速率为5000rpm，离心时间为20分钟，得到上清液，采用烘干称重法测得上清液干物质含量为2％。The obtained thermally extracted product is subjected to centrifugation, wherein the centrifugation speed is 5000 rpm and the centrifugation time is 20 minutes to obtain a supernatant, and the dry matter content of the supernatant is 2% measured by drying weighing method.
(3)酶解：取步骤(2)所得上清液45g，添加0.027g维生素C和0.018g木瓜蛋白酶，在30℃、pH值8.0条件下酶解8h；之后添加0.018g风味蛋白酶，在30℃、pH值8的条件下酶解12h。(3) Enzymatic hydrolysis: take 45 g of the supernatant obtained in step (2), add 0.027 g of vitamin C and 0.018 g of papain, and enzymatically hydrolyze 8 h at 30°C and pH 8.0; then add 0.018 g of flavor protease, and at 30 The enzymatic hydrolysis was carried out under the conditions of ℃ and pH 8 for 12 h.
(4)将步骤(3)酶解后的产物进行喷雾干燥，得到富含半胱氨酸的酵母抽提物，其中喷雾干燥进风温度控制在155-175℃，出风温度控制在90-115℃，雾化压力控制在100-160bar，收料温度控制在25-45℃。(4) spray-drying the product after enzymolysis in step (3) to obtain a cysteine-rich yeast extract, wherein the spray-drying inlet air temperature is controlled at 155-175°C, and the outlet air temperature is controlled at 90- 115℃, the atomization pressure is controlled at 100-160bar, and the temperature of the material is controlled at 25-45℃.
实施例产物检测：Example product detection:
为了验证本发明上述实施例1-6以及对比例1-4的产物性能，采取高效液相色谱法进行酵母抽提物中的半胱氨酸含量测试，检测方法如下：In order to verify the product properties of the above-mentioned embodiments 1-6 and comparative examples 1-4 of the present invention, the cysteine content in the yeast extract is tested by high performance liquid chromatography, and the detection method is as follows:
1.取1g样品溶于5ml水，向其中加入100微升，质量浓度为5％的苯酚，之后加入10ml，6mol/L的浓盐酸，放置两个小时后，取100微升定容至10ml。1. Take 1g of sample and dissolve it in 5ml of water, add 100μl of phenol with a mass concentration of 5%, then add 10ml of 6mol/L concentrated hydrochloric acid, leave it for two hours, take 100μl and dilute to 10ml .
2.取0.01克半胱氨酸标准品溶于10ml水后，再取200微升，400微升，600微升，800微升，1毫升，定容至1毫升。2. Dissolve 0.01 g of cysteine standard in 10 ml of water, then take 200 microliters, 400 microliters, 600 microliters, 800 microliters, 1 ml, and dilute to 1 ml.
3.色谱条件3. Chromatographic conditions
色谱柱：安捷伦SB-C18柱。Chromatographic column: Agilent SB-C18 column.
流动相：A相是0.1％磷酸，B相是乙醇，按80:20比例，梯度平衡3-4小时。Mobile phase: phase A is 0.1% phosphoric acid, phase B is ethanol, in a ratio of 80:20, and the gradient is equilibrated for 3-4 hours.
柱温箱温度：25℃。Column oven temperature: 25°C.
流速 0.6mL/min。The flow rate is 0.6mL/min.
检测波长：205nm。Detection wavelength: 205nm.
测定时间：7分钟。Measurement time: 7 minutes.
4.测定4. Determination
将标准系列和样品溶液分别进样，进样量为0.6uL，根据标准品的保留时间定性半胱氨酸的色谱峰并绘制标准曲线(标准品浓度和峰面积的关系曲线)，根据样品的峰面积，以外标法计算半胱氨酸的百分含量。The standard series and the sample solution were injected separately, and the injection volume was 0.6uL. The chromatographic peak of cysteine was qualitatively determined according to the retention time of the standard and the standard curve (the relationship curve between the concentration of the standard and the peak area) was drawn. Peak area, and the percentage of cysteine was calculated by the external standard method.
5.结果计算5. Result calculation
X＝(C*V)/(M*(100-X1))/10000*100％X=(C*V)/(M*(100-X1))/10000*100%
X——样品中半胱氨酸的含量，％；X——the content of cysteine in the sample, %;
M——样品的质量，gM - the mass of the sample, g
X1——样品中水分的含量,％；X1——Moisture content in the sample, %;
C——根据样品峰面积查标准曲线的浓度，ug/mLC——Check the concentration of the standard curve according to the peak area of the sample, ug/mL
V——样品定容体积，mLV——Sample volume, mL
检测结果如表2所示。The test results are shown in Table 2.
表2 酵母抽提物中半胱氨酸含量Table 2 Cysteine content in yeast extract
Figure PCTCN2021137074-appb-000001
Figure PCTCN2021137074-appb-000001
由表2可知，本发明富含半胱氨酸的酵母抽提物制备方法可制得半胱氨酸含量大于等于1％的的酵母抽提物。As can be seen from Table 2, the cysteine-rich yeast extract preparation method of the present invention can obtain yeast extracts with a cysteine content greater than or equal to 1%.
以上所述，仅是本发明实施的较佳实施例，并非对本发明做任何形式上的限制，凡在本发明的精神和原则之内所做的修改、等同替换和改进等，均需要包含在本发明的保护范围之内。The above are only preferred embodiments of the present invention, and are not intended to limit the present invention in any form. Any modifications, equivalent replacements and improvements made within the spirit and principles of the present invention shall be included in the within the protection scope of the present invention.

Claims (14)

  1. 一种富含半胱氨酸的酵母抽提物，其特征在于，所述酵母抽提物通过包含下述步骤制备得到：A cysteine-rich yeast extract, characterized in that, the yeast extract is prepared by comprising the following steps:
    (1)将原始菌株发酵得到酵母乳；(1) the original strain is fermented to obtain yeast milk;
    (2)将步骤(1)所得酵母乳热提取、离心得到上清液；(2) the yeast milk obtained in step (1) is thermally extracted and centrifuged to obtain a supernatant;
    (3)将步骤(2)所得上清液酶解，得到富含半胱氨酸的酵母抽提物；(3) enzymolysis of the supernatant obtained in step (2) to obtain a cysteine-rich yeast extract;
    其中，步骤(1)中所述原始菌株为富含还原型谷胱甘肽的酵母菌株(saccharomyces cerevisiae)，保藏编号为：CCTCC M 205130，保藏地址为：于2005年10月25日保藏于中国典型培养物保藏中心(武汉)。Wherein, the original strain described in the step (1) is a yeast strain (saccharomyces cerevisiae) rich in reduced glutathione, the deposit number is: CCTCC M 205130, and the deposit address is: on October 25, 2005, it was preserved in China Type Culture Collection (Wuhan).
  2. 根据权利要求1所述酵母抽提物，其特征在于，以酵母抽提物干物质的质量计，酵母抽提物中半胱氨酸含量≥1％，优选为所述半胱氨酸含量≥2％。The yeast extract according to claim 1, wherein the cysteine content in the yeast extract is greater than or equal to 1% based on the mass of the dry matter of the yeast extract, preferably the cysteine content is greater than or equal to 1%. 2%.
  3. 根据权利要求1或2所述酵母抽提物，其特征在于，步骤(1)中所述发酵过程以补料流加方式添加营养液；优选的，发酵过程包含普通培养过程和饥饿培养过程，按重量百份比计，饥饿培养过程营养液添加流量为普通培养过程添加流量的10-40％。The yeast extract according to claim 1 or 2, wherein the fermentation process described in the step (1) adds nutrient solution in a fed-feed mode; preferably, the fermentation process comprises a common culture process and a starvation culture process, In terms of weight percentage, the addition flow rate of the nutrient solution in the starvation culture process is 10-40% of the addition flow rate in the normal culture process.
  4. 根据权利要求3所述酵母抽提物，其特征在于，所述普通培养的温度为25-35℃，优选普通培养的时间为5-20h；进一步优选的，所述饥饿培养的温度为25-35℃，优选饥饿培养的时间为5-10h。The yeast extract according to claim 3, wherein the temperature of the ordinary culture is 25-35°C, preferably the time of the ordinary culture is 5-20h; further preferably, the temperature of the starvation culture is 25-20h 35°C, preferably the time of starvation culture is 5-10h.
  5. 根据权利要求1-4任一项所述酵母抽提物，其特征在于，步骤(2)热提取酵母乳的质量浓度为8-15％；优选热提取的温度为80-110℃，进一步优选为95℃；优选热提取时间为60-120min，进一步优选为90min；优选热提取pH值为5.0-8.0。The yeast extract according to any one of claims 1-4, characterized in that, in step (2), the mass concentration of the thermally extracted yeast milk is 8-15%; preferably, the temperature of thermal extraction is 80-110° C., more preferably The temperature is 95°C; the preferred thermal extraction time is 60-120 min, more preferably 90 min; the preferred thermal extraction pH value is 5.0-8.0.
  6. 根据权利要求1-5任一项所述酵母抽提物，其特征在于，步骤(3)酶解时，依次采用内切蛋白酶和外切蛋白酶进行酶解，优选所述内切蛋白酶选自碱性蛋白酶、木瓜蛋白酶、胰蛋白酶和中性蛋白酶中的一种或两种以上；进一步优选所述外切蛋白酶选自风味蛋白酶、羧肽酶和氨肽酶中的一种或两种以上。The yeast extract according to any one of claims 1-5, characterized in that, during the enzymatic hydrolysis in step (3), endoprotease and exoprotease are used for enzymolysis in sequence, and preferably the endoprotease is selected from alkali One or more of the protease, papain, trypsin and neutral protease; further preferably, the exoprotease is selected from one or more of the flavor protease, carboxypeptidase and aminopeptidase.
  7. 根据权利要求6所述酵母抽提物，其特征在于，以上清液干物质的质 量计，所述内切蛋白酶的添加量为0.1-1.5％，优选为1％。The yeast extract according to claim 6, characterized in that, based on the mass of the supernatant dry matter, the addition amount of the endoprotease is 0.1-1.5%, preferably 1%.
  8. 根据权利要求6或7所述酵母抽提物，其特征在于，内切蛋白酶酶解时，酶解温度为40-80℃，优选的，酶解pH值为4.0-7.5，酶解时间为10-15h。The yeast extract according to claim 6 or 7, characterized in that, during enzymatic hydrolysis by endoprotease, the enzymatic hydrolysis temperature is 40-80° C., preferably, the enzymatic hydrolysis pH value is 4.0-7.5, and the enzymatic hydrolysis time is 10 -15h.
  9. 根据权利要求6-8任一项所述酵母抽提物，其特征在于，以上清液干物质的质量计，所述外切蛋白酶的添加量为0.1-1.5％，优选为1％。The yeast extract according to any one of claims 6-8, characterized in that, based on the mass of the dry matter of the supernatant, the addition amount of the exoprotease is 0.1-1.5%, preferably 1%.
  10. 根据权利要求6-9任一项所述酵母抽提物，其特征在于，外切蛋白酶酶解时，酶解温度为40-80℃；优选的，酶解pH值为4.0-7.5，酶解时间为5-10h。The yeast extract according to any one of claims 6-9, characterized in that, during enzymatic hydrolysis by exoprotease, the enzymatic hydrolysis temperature is 40-80° C.; The time is 5-10h.
  11. 根据权利要求1-10中任一项所述酵母抽提物，其特征在于，步骤(3)酶解前还包括：向步骤(2)得到的上清液中加入维生素C；优选的，以上清液干物质的质量计，所述维生素C的添加量为0.5-2％；进一步优选为1％。The yeast extract according to any one of claims 1-10, characterized in that, before step (3) enzymatic hydrolysis, further comprising: adding vitamin C to the supernatant obtained in step (2); preferably, the above In terms of the mass of the dry matter of the clear liquid, the addition amount of the vitamin C is 0.5-2%; more preferably, it is 1%.
  12. 根据权利要求1-11中任一项所述酵母抽提物，其特征在于，步骤(3)后还包括下述步骤：将步骤(3)酶解后酶解液依次进行蒸发浓缩和干燥，得到富含半胱氨酸的酵母抽提物。The yeast extract according to any one of claims 1-11, characterized in that, after step (3), it further comprises the following steps: the enzymatic hydrolysis solution after the enzymatic hydrolysis in step (3) is successively evaporated, concentrated and dried, A cysteine-rich yeast extract is obtained.
  13. 根据权利要求12所述酵母抽提物，其特征在于，所述干燥为喷雾干燥、真空干燥或滚筒干燥中的一种,优选为喷雾干燥。The yeast extract according to claim 12, wherein the drying is one of spray drying, vacuum drying or drum drying, preferably spray drying.
  14. 权利要求1-13任一项所述酵母抽提物的制备方法，其特征在于，包括以下步骤：The preparation method of the yeast extract described in any one of claims 1-13, is characterized in that, comprises the following steps:
    (1)将原始菌株发酵得到酵母乳；(1) the original strain is fermented to obtain yeast milk;
    (2)将步骤(1)所得酵母乳热提取、离心得到上清液；(2) the yeast milk obtained in step (1) is thermally extracted and centrifuged to obtain a supernatant;
    (3)将步骤(2)所得上清液酶解，得到富含半胱氨酸的酵母抽提物；(3) enzymolysis of the supernatant obtained in step (2) to obtain a cysteine-rich yeast extract;
    其中，步骤(1)中所述原始菌株为富含还原型谷胱甘肽的酵母菌株((saccharomyces cerevisiae)，保藏编号为：CCTCC M 205130，保藏地址为：于2005年10月25日保藏于中国典型培养物保藏中心(武汉)。Wherein, the original strain described in the step (1) is a yeast strain ((saccharomyces cerevisiae) rich in reduced glutathione, the deposit number is: CCTCC M 205130, and the deposit address is: on October 25, 2005, it was preserved in China Type Culture Collection (Wuhan).
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