CN112998152B - Yeast hydrolysate and preparation method and application thereof - Google Patents
Yeast hydrolysate and preparation method and application thereof Download PDFInfo
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- CN112998152B CN112998152B CN201911307412.9A CN201911307412A CN112998152B CN 112998152 B CN112998152 B CN 112998152B CN 201911307412 A CN201911307412 A CN 201911307412A CN 112998152 B CN112998152 B CN 112998152B
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/153—Nucleic acids; Hydrolysis products or derivatives thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Animal Husbandry (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Birds (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Physiology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fodder In General (AREA)
Abstract
The invention relates to the field of biological fermentation, in particular to a yeast hydrolysate and a preparation method and application thereof. The invention provides a yeast hydrolysate which is characterized by comprising the following components in parts by weight: the total amount of the mannooligosaccharide and the beta-glucan is 30-50%, the nucleic acid is 4-8%, and the crude protein is 35-50%. The nucleic acid, protein, mannooligosaccharide and beta-glucan which are rich in the yeast hydrolysate can provide essential nutritional ingredients for the growth of the chicks, can improve the intestinal development and the growth speed of the chicks, can improve the immune function of the chicks and can protect the health of young birds.
Description
Technical Field
The invention relates to the field of biological fermentation, in particular to a yeast hydrolysate and a preparation method and application thereof.
Background
With the vigorous development of the farming industry, early nutrition of poultry is becoming more and more interesting because the growth and development of young stage is directly related to the subsequent growth of poultry. The weight gain of the poultry after the shell is removed for one week is improved, and the time of the poultry to market can be obviously shortened. Promoting the growth and development of the chicks, enhancing the immunity protection of the chicks and improving the health of the chicks, and becoming a focus of attention for poultry farming.
Currently, most of yeast products related to poultry feeds are yeast cultures, brewers' yeast and fermentation waste streams. The yeast culture is obtained by fermenting and culturing yeast fermentation metabolic products and an metamorphosis culture medium; saccharomyces cerevisiae is a by-product of brewer's yeast; the fermentation waste liquid belongs to the deep processing treatment of industrial waste liquid, the quality of the product is unstable, and the improvement effect on the growth of poultry is not obvious. The yeast hydrolysate is a stable, safe and high-quality single-cell protein raw material obtained by deep enzymolysis of pure yeast milk, and nucleic acid, protein, mannooligosaccharide and beta-glucan contained in the yeast hydrolysate are nutrients necessary for young poultry, so that the yeast hydrolysate has good effect of improving intestinal development, immune function and growth speed of poultry.
The yeast hydrolysate is rich in nutrients, and contains multiple essential amino acids, vitamins, minerals, nucleic acids, beta-glucan, and mannooligosaccharide. The beta-glucan can stimulate animal body to produce macrophage with critical action on immunity, and can eliminate pathogenic microbe with damaged and decayed cell nucleus invaded into body. The mannooligosaccharide can promote the growth of certain beneficial bacteria, inhibit intestinal pathogenic bacteria, improve the total amount of volatile fatty acid in intestinal tracts, and enhance the immunity of organisms by improving the T lymphocyte conversion rate, the macrophage phagocytosis index, the avian influenza antibody titer and the like.
Disclosure of Invention
The technical problems solved by the invention are as follows: at present, the types of yeast hydrolysates are various, and the quality of products is uneven. The invention aims to provide a special yeast hydrolysate for chicken feed, which can have good improving effects on intestinal development, immune function and growth speed of chicken.
The yeast hydrolysate is widely popularized and used in the production of chicken feeds, so that the shortage of protein resources for feeding can be relieved, and the application prospect is very broad.
Specifically, the invention provides the following technical scheme.
The invention provides a yeast hydrolysate which comprises the following components in percentage by mass: the total amount of the mannooligosaccharide and the beta-glucan is 30-50%, the nucleic acid is 4-8%, and the crude protein is 35-50%.
Preferably, the total amount of the mannooligosaccharide and the beta-glucan is 31.5-50%, the nucleic acid is 5-7% and the crude protein is 35-45% by mass percent.
Preferably, wherein the ratio of the content of the mannooligosaccharide to the content of the beta-glucan is 1:4-4:1, and preferably the ratio of the content of the mannooligosaccharide to the content of the beta-glucan is 1:1.2.
The invention also provides a preparation method of the yeast hydrolysate, which comprises the following steps:
(1) And (3) fermenting and culturing yeast: fermenting and culturing a yeast strain to obtain yeast milk;
(2) Autolysis of yeast: autolysis is carried out on the yeast milk obtained in the step (1);
(3) And (3) yeast enzymolysis: adding a complex enzyme preparation into the product obtained in the step (2), and carrying out enzymolysis to obtain yeast hydrolysate;
the complex enzyme preparation is one or more than two of papain, alkaline protease, neutral protease and flavourzyme.
Preferably, wherein the yeast strain is selected from Saccharomyces cerevisiae FX-2 (Saccharomyces cerevisiae FX-2) and/or Saccharomyces cerevisiae Z2.1 (Saccharomyces cerevisiae Hansen Z2.1), preferably Saccharomyces cerevisiae FX-2 (Saccharomyces cerevisiaeFX-2).
Preferably, the yeast fermentation culture in the step (1) adopts a culture medium containing a carbon source, a nitrogen source and a phosphorus source;
preferably, the carbon source accounts for 80-90% of the culture medium, the nitrogen source accounts for 10-15% of the culture medium, and the phosphorus source accounts for 1-5% of the culture medium;
more preferably, the carbon source is added in an amount of 85% by mass of the medium, the nitrogen source is added in an amount of 12% by mass of the medium, and the phosphorus source is added in an amount of 3% by mass of the medium.
Preferably, the carbon source is molasses and/or starch hydrolysis sugar, the nitrogen source is ammonia water and/or ammonium sulfate, and the phosphorus source is phosphoric acid;
preferably, the carbon source is molasses and the nitrogen source is ammonia water.
Preferably, the fermentation temperature in the step (1) is 25-35 ℃, preferably 29-32 ℃, the fermentation pH is 5.0-7.0, preferably 5.0-6.0, and the fermentation time is 15-18 hours.
Preferably, in the step (2), citric acid accounting for 1-5%o of the culture medium is added according to the mass percentage.
Preferably, the method further comprises a process of heat-shock the yeast milk obtained in the step (1) at 85-95 ℃ after the step (1) and before the step (2).
Preferably, in the step (3), 4-20%o of the complex enzyme preparation is added in percentage by mass;
preferably, the content of papain in the compound enzyme preparation is 1-5%o, the content of alkaline protease is 1-5%o, the content of neutral protease is 1-5%o, and the content of flavourzyme is 1-5%o.
Preferably, the enzymolysis time in the step (3) is 6-12 hours, preferably 10-12 hours.
Preferably, the method further comprises the step of drying the product obtained in the step (3) at 70-80 ℃ for 1-2 h.
The invention also provides a feed comprising the yeast hydrolysate.
Preferably, the feed, wherein the addition amount of yeast hydrolysate in the feed is 1-50kg/t.
Preferably, when the chicks are meat chicks, the addition amount of the yeast hydrolysate in the feed is 1-20kg/t; preferably 5kg/t;
when the chicks are egg chicks, the yeast hydrolysate is added to the feed in an amount of 5-50kg/t, preferably 5-10kg/t.
The invention also provides an application of the yeast hydrolysate in the field of feed, preferably in the field of chicken feed, and more preferably in the field of chicken feed.
The beneficial effects obtained by the invention are as follows:
the yeast hydrolysate can be used as a feed raw material, is rich in protein, amino acid, small peptide, polysaccharide, vitamin and other components, and can provide essential nutritional components for the growth of chickens. Animal experiments prove that the yeast hydrolysate has the advantages that the yeast hydrolysate is used as the feed for the chicks, can provide sufficient and balanced nutritional ingredients for the chicks, can improve intestinal development and the growth speed of the chicks, can improve the immune function of the chicks, and can protect the health of young birds.
Strain preservation information
The strain Saccharomyces cerevisiae FX-2 (Saccharomyces cerevisiae FX-2) used in the invention is preserved in China Center for Type Culture Collection (CCTCC) at 8-1-2016, and the preservation number is CCTCC NO: M2016418, and the preservation address is: chinese, wuhan, university of Wuhan, postal code: 430072; telephone: (027) 68754052, which strain is described in the patent publication No. CN 201611141122.8.
The saccharomyces cerevisiae strain Z2.1 (Saccharomyces cerevisiae Hansen Z2.1) used in the invention is preserved in China center for type culture collection (China center for type culture collection) in 10 and 25 days 2005, and the strain has a preservation number of CCTCC NO: M205127 and a preservation address of: chinese, wuhan, university of Wuhan, post code: 430072, telephone: (027) 68752319, this strain is described in the patent publication No. CN 201710522840.8.
Detailed Description
As described above, the invention provides a yeast hydrolysate, which comprises the following components in percentage by mass: the total amount of the mannooligosaccharide and the beta-glucan is 30-50%, the nucleic acid is 4-8%, and the crude protein is 35-50%.
Preferably, the total amount of the mannooligosaccharide and the beta-glucan is 31.5-50%, the nucleic acid is 5-7% and the crude protein is 35-45% by mass percent.
Preferably, the ratio of the content of the mannooligosaccharide to the content of the beta-glucan is 1:4-4:1, and preferably the ratio of the content of the mannooligosaccharide to the content of the beta-glucan is 1:1.2.
The invention also provides a preparation method of the yeast hydrolysate, which comprises the following steps:
(1) And (3) fermenting and culturing yeast: fermenting and culturing a yeast strain to obtain yeast milk;
(2) Autolysis of yeast: autolysis is carried out on the yeast milk obtained in the step (1);
(3) And (3) yeast enzymolysis: adding a complex enzyme preparation into the product obtained in the step (2), and carrying out enzymolysis to obtain yeast hydrolysate;
the complex enzyme preparation is one or more than two of papain, alkaline protease, neutral protease and flavourzyme.
Preferably, wherein the yeast strain is selected from Saccharomyces cerevisiae FX-2 (Saccharomyces cerevisiae FX-2) and/or Saccharomyces cerevisiae Z2.1 (Saccharomyces cerevisiae Hansen Z2.1), preferably Saccharomyces cerevisiae FX-2 (Saccharomyces cerevisiae FX-2).
Preferably, the yeast fermentation culture in the step (1) adopts a culture medium containing a carbon source, a nitrogen source and a phosphorus source;
preferably, the carbon source accounts for 80-90% of the culture medium, the nitrogen source accounts for 10-15% of the culture medium, and the phosphorus source accounts for 1-5% of the culture medium;
more preferably, the carbon source is added in an amount of 85% by mass of the medium, the nitrogen source is added in an amount of 12% by mass of the medium, and the phosphorus source is added in an amount of 3% by mass of the medium.
Preferably, the carbon source is molasses and/or starch hydrolysis sugar, the nitrogen source is ammonia water and/or ammonium sulfate, and the phosphorus source is phosphoric acid;
preferably, the carbon source is molasses and the nitrogen source is ammonia water.
Preferably, the fermentation temperature in the step (1) is 25-35 ℃, preferably 29-32 ℃, the fermentation pH is 5.0-7.0, preferably 5.0-6.0, and the fermentation time is 15-18 hours.
Preferably, in the step (2), citric acid accounting for 1-5%o of the culture medium is added according to the mass percentage.
Preferably, the method further comprises a process of heat-shock the yeast milk obtained in the step (1) at 85-95 ℃ after the step (1) and before the step (2).
Preferably, in the step (3), 4-20%o of the complex enzyme preparation is added in percentage by mass;
preferably, the content of papain in the compound enzyme preparation is 1-5%o, the content of alkaline protease is 1-5%o, the content of neutral protease is 1-5%o, and the content of flavourzyme is 1-5%o.
Preferably, the enzymolysis time in the step (3) is 6-12 hours, preferably 10-12 hours.
Preferably, the method further comprises the step of drying the product obtained in the step (3) at 70-80 ℃ for 1-2 h.
The invention also provides a feed comprising the yeast hydrolysate.
Preferably, the feed, wherein the addition amount of yeast hydrolysate in the feed is 1-50kg/t.
Preferably, when the chicks are meat chicks, the addition amount of the yeast hydrolysate in the feed is 1-20kg/t; preferably 5kg/t; when the chicks are egg chicks, the yeast hydrolysate is added to the feed in an amount of 5-50kg/t, preferably 5-10kg/t.
The invention also provides an application of the yeast hydrolysate in the field of feed, preferably in the field of chicken feed, and more preferably in the field of chicken feed.
The raw materials and equipment manufacturers used in this example, as well as the equipment and analysis methods used in the analysis of the products, are described below, wherein the chemicals are not identified as chemically pure grades of conventional reagents. The information on the raw materials used in the examples and the experimental equipment are shown in table 1.
Table 1 of information on raw materials and equipment used in examples
| Raw material information | Purity/model | Manufacturing factories |
| Molasses | Chifeng blue sky sugar industry Co.Ltd | |
| Starch hydrolyzing sugar | Food grade | HUANGLONG FOOD INDUSTRY Co.,Ltd. |
| Citric acid | Analytical grade | Yingxuan Co., ltd |
| Ammonia water | Analytical grade | Fang national Ann chemical Co., ltd |
| Ammonium sulfate | Analytical grade | Jiangsu Koronto Co Ltd |
| Phosphoric acid | Analytical grade | Tianjin Dingshengxin chemical industry Co.Ltd |
| Papain | 60 ten thousand U/g | Wohan Borui Source Biotechnology Co.Ltd |
| Neutral protease | 30 ten thousand U/g | NINGXIA SUNSON INDUSTRY GROUP Co.,Ltd. |
| Alkaline protease | 50 ten thousand U/g | NANNING PANGBO BIOLOGICAL ENGINEERING Co.,Ltd. |
| Flavoured protease | Indonesia Germany | |
| Ultra-high temperature instantaneous sterilizer | RSCG01-1 | Wenzhou city long macro light industry machinery Co.Ltd |
The microbial cells of Saccharomyces cerevisiae (Saccharomyces cerevisiae FX-2) used in the examples and comparative examples of the present invention were prepared by performing an expansion culture by the following method:
1) Activating strains: the colony is picked, placed in YPD liquid culture medium, placed in a shaking table to activate strain.
2) And (3) strain purification: the bacterial liquid after the activation in the last step is diluted and plated in a gradient way to obtain single bacterial colonies.
3) And (3) strain expansion culture: single colony in the last step plate is picked and inoculated to 200mLYPD liquid culture medium, and cultured for 18 hours at the temperature of 30 ℃ and the rotating speed of 220 rpm; secondary seed culture, namely inoculating the primary cultured seeds into 1L of secondary culture medium according to 10% of the volume of the secondary culture medium, and culturing for 18 hours at the temperature of 28 ℃ and the rotating speed of 200 rpm; the pH was controlled at 5.5.
4) Seed enrichment: centrifuging at 5000rpm, washing with deionized water with low calcium ion and magnesium ion content for 3 times, and concentrating to obtain Saccharomyces cerevisiae seed until the wet weight of yeast is 220g/L.
The cells of Saccharomyces cerevisiae (Saccharomyces cerevisiae Hansen Z2.1) used in the examples and comparative examples of the present invention were prepared by performing an expansion culture by the following method:
1) Activating strains: the colony is picked, placed in YPD liquid culture medium, placed in a shaking table to activate strain.
2) And (3) strain purification: the bacterial liquid after the activation in the last step is diluted and plated in a gradient way to obtain single bacterial colonies.
3) And (3) strain expansion culture: single colony in the last step plate is picked and inoculated to 200mLYPD liquid culture medium, and cultured for 18 hours at the temperature of 30 ℃ and the rotating speed of 220 rpm; secondary seed culture, namely inoculating the primary cultured seeds into 1L of secondary culture medium according to 10% of the volume of the secondary culture medium, and culturing for 18 hours at the temperature of 28 ℃ and the rotating speed of 200 rpm; the pH was controlled at 5.5.
4) Seed enrichment: centrifuging at 5000rpm, washing with deionized water with low calcium ion and magnesium ion content for 3 times, and concentrating to obtain Saccharomyces cerevisiae seed until the wet weight of yeast is 220g/L.
In examples and comparative examples, the determination of the protein content in yeast hydrolysates is referred to GB/T6432-94; determination of nucleic acid content reference spectrophotometry for determination of total phosphorus in feed (GB/T6437) determination, determination of mannooligosaccharides and beta-glucan was performed by liquid chromatography.
EXAMPLE one preparation of Yeast hydrolysates
(1) Yeast fermentation culture
Preparing a culture solution, wherein the culture solution contains a carbon source, a nitrogen source and a phosphorus source, the carbon source is molasses and starch hydrolysis sugar, the mass of the molasses is 4000g, and the concentration of the molasses is 35% (mass); 2000g of starch hydrolyzing sugar, the concentration of which is 40% by mass. The nitrogen source is ammonia water and ammonium sulfate, wherein the mass of the ammonia water is 400g, and the concentration of the ammonia water is 17% (mass); the mass of ammonium sulfate was 300g, the phosphorus source was phosphoric acid, the mass thereof was 750g, and the concentration thereof was 8% by mass. Sterilizing at 120deg.C for 8min, feeding into a tank, inoculating 50g of strain of Saccharomyces cerevisiae FX-2, controlling fermentation temperature at 32deg.C, fermenting for 15 hr, and fermenting at pH of 6.0 to obtain pure culture liquid fermented yeast milk.
(2) Autolysis of yeast
Performing heat shock on yeast milk at 90 ℃ for 40 seconds, adding 2.5%o citric acid into the yeast milk for adjustment, heating to 50 ℃, and keeping the temperature at 50 ℃ for autolysis for 8 hours;
(3) And (3) yeast enzymolysis:
the pH of the solution after autolysis treatment is adjusted to 5.5, the temperature is 60 ℃, and a compound enzyme preparation containing 2.5 permillage papain, 2.5 permillage alkaline protease, 2.5 permillage neutral protease and 2.5 permillage flavourzyme is added for enzymolysis reaction for 8 hours according to the dry matter of yeast milk.
After the enzymolysis reaction is finished, heating to 75 ℃, preserving heat for 1.5 hours, inactivating enzymes, concentrating and drying to obtain a yeast hydrolysate product A.
And measuring the protein content, the nucleic acid content and the content of the mannooligosaccharide and the beta-glucan of the yeast hydrolysate product A prepared by the method. Wherein the protein content is 42%, the nucleic acid content is 6%, the content of the obtained mannooligosaccharide and beta-glucan is 40%, the content of the mannooligosaccharide is 18.5%, and the content of the beta-glucan is 21.5%.
EXAMPLE two preparation of Yeast hydrolysates
(1) Yeast fermentation culture
Preparing a culture solution, wherein the culture solution contains a carbon source, a nitrogen source and a phosphorus source, the carbon source is molasses and starch hydrolysis sugar, the mass of the molasses is 3000g, and the concentration of the molasses is 45% (mass); 3000g of starch hydrolyzing sugar, the concentration of which is 30% by mass. The nitrogen source is ammonia water and ammonium sulfate, wherein the mass of the ammonia water is 450g, and the concentration of the ammonia water is 15 percent (mass); the mass of ammonium sulfate was 200g, the phosphorus source was phosphoric acid, the mass thereof was 700g, and the concentration thereof was 10% by mass. Sterilizing at 120deg.C for 8min, feeding into a tank, inoculating 50g of strain of Saccharomyces cerevisiae FX-2, controlling the fermentation temperature at 29 deg.C, fermenting for 18 hr, and fermenting at pH of 7.0 to obtain pure culture liquid fermented yeast milk.
(2) Autolysis of yeast
Performing heat shock on yeast milk at the temperature of 85 ℃ for 40 seconds, adding 1%o citric acid into the yeast milk for adjustment, heating to 45 ℃, and keeping the temperature at 45 ℃ for autolysis for 12 hours;
(3) And (3) yeast enzymolysis:
the pH value of the solution after autolysis treatment is regulated to 6.0, the temperature is 55 ℃, and a compound enzyme preparation containing 1 permillage papain, 1 permillage alkaline protease, 1 permillage neutral protease and 1 permillage flavourzyme is added for enzymolysis reaction for 12 hours according to the dry matter of yeast milk.
After the enzymolysis reaction is finished, heating to 80 ℃, preserving heat for 2 hours, inactivating enzymes, concentrating and drying to obtain a yeast hydrolysate product B.
And (3) measuring the protein content, the nucleic acid content and the content of the mannooligosaccharide and the beta-glucan of the yeast hydrolysate product B. Wherein the protein content is 35%, the nucleic acid content is 5%, the content of the obtained mannooligosaccharide and beta-glucan is 50%, the content of the mannooligosaccharide is 22.5%, and the content of the beta-glucan is 27.5%.
EXAMPLE three preparation of Yeast hydrolysates
(1) Yeast fermentation culture
Preparing a culture solution, wherein the culture solution contains a carbon source, a nitrogen source and a phosphorus source, the carbon source is molasses and starch hydrolysis sugar, the mass of the molasses is 3500g, and the concentration of the molasses is 40% (mass); 2500g of starch hydrolyzing sugar, the concentration of which is 35% by mass. The nitrogen source is ammonia water and ammonium sulfate, wherein the mass of the ammonia water is 430g, and the concentration of the ammonia water is 16% (mass); the mass of ammonium sulfate was 250g, the phosphorus source was phosphoric acid, the mass thereof was 730g, and the concentration thereof was 9% by mass. Sterilizing at 120deg.C for 8min, feeding into a tank, inoculating 50g of strain of Saccharomyces cerevisiae FX-2, controlling fermentation temperature at 32deg.C, fermenting for 12 hr, and fermenting at pH of 5.0 to obtain pure culture liquid fermented yeast milk.
(2) Autolysis of yeast
Performing heat shock on yeast milk at 95 ℃ for 40 seconds, adding 5%o citric acid into the yeast milk for adjustment, heating to 55 ℃, and keeping the temperature at 55 ℃ for autolysis for 6 hours;
(3) And (3) yeast enzymolysis:
the pH value of the solution after autolysis treatment is regulated to 4.0, the temperature is 65 ℃, and a compound enzyme preparation containing 5 permillage papain, 5 permillage alkaline protease, 5 permillage neutral protease and 5 permillage flavourzyme is added for enzymolysis reaction for 6 hours according to the dry matter of yeast milk.
After the enzymolysis reaction is finished, heating to 70 ℃, preserving heat for 1h, inactivating enzyme, concentrating and drying to obtain a yeast hydrolysate product C.
And measuring the protein content, the nucleic acid content and the content of the mannooligosaccharide and the beta-glucan of the yeast hydrolysate product C. Wherein the protein content is 45%, the nucleic acid content is 7%, the content of the obtained mannooligosaccharide and beta-glucan is 31.5%, the content of the mannooligosaccharide is 14.9%, and the content of the beta-glucan is 16.6%.
Comparative example preparation of Yeast hydrolysate
(1) Yeast fermentation culture
Preparing a culture solution, wherein the culture solution contains a carbon source, a nitrogen source and a phosphorus source, the carbon source is molasses and starch hydrolysis sugar, the mass of the molasses is 6000g, and the concentration of the molasses is 40% (mass); the nitrogen source is ammonia water and ammonium sulfate, wherein the mass of the ammonia water is 400g, and the concentration of the ammonia water is 17% (mass); the mass of ammonium sulfate was 300g, the phosphorus source was phosphoric acid, the mass thereof was 750g, and the concentration thereof was 10% by mass. Sterilizing at 120deg.C for 8min, feeding into a tank, inoculating 50g of Saccharomyces cerevisiae Z2.1, controlling fermentation temperature to 30deg.C, fermenting for 15 hr, and fermenting at pH of 5.6 to obtain pure culture liquid fermented yeast milk.
(2) Autolysis of yeast
Performing heat shock on yeast milk at 90 ℃ for 40 seconds, adding 2.5%o citric acid into the yeast milk for adjustment, heating to 50 ℃, and keeping the temperature at 50 ℃ for autolysis for 8 hours;
(3) And (3) yeast enzymolysis:
the pH value of the solution after autolysis treatment is adjusted to 5.0, the temperature is 60 ℃, and a compound enzyme preparation containing 2.5 permillage papain, 2.5 permillage alkaline protease, 2.5 permillage neutral protease and 2.5 permillage flavourzyme is added for enzymolysis reaction for 8 hours according to the dry matter of yeast milk.
After the enzymolysis reaction is finished, heating to 75 ℃, preserving heat for 1.5 hours, inactivating enzymes, concentrating and drying to obtain a yeast hydrolysate product D.
And measuring the protein content, the nucleic acid content and the content of the mannooligosaccharide and the beta-glucan of the yeast hydrolysate product D. Wherein the protein content is 51%, the nucleic acid content is 10%, the content of the obtained mannooligosaccharide and beta-glucan is 27.3%, the content of the mannooligosaccharide is 12.8%, and the content of the beta-glucan is 14.5%.
Comparative example preparation of a Yeast hydrolysate
(1) Yeast fermentation culture
Preparing a culture solution, wherein the culture solution contains a carbon source, a nitrogen source and a phosphorus source, the carbon source is molasses and starch hydrolysis sugar, the mass of the molasses is 4000g, and the concentration of the molasses is 35% (mass); 2000g of starch hydrolyzing sugar, the concentration of which is 40% by mass. The nitrogen source is ammonia water and ammonium sulfate, wherein the mass of the ammonia water is 400g, and the concentration of the ammonia water is 17% (mass); the mass of ammonium sulfate was 300g of phosphoric acid as a phosphorus source, the mass thereof was 750g, and the concentration thereof was 8% by mass. Sterilizing at 120deg.C for 8min, feeding into a tank, inoculating 50g of strain of Saccharomyces cerevisiae FX-2, fermenting at 30deg.C for 8 hr, and fermenting at pH of 6.0 to obtain pure culture liquid fermented yeast milk.
(2) Autolysis of yeast
Performing heat shock on yeast milk at 90 ℃ for 40 seconds, adding 2.5%o citric acid into the yeast milk for adjustment, heating to 50 ℃, and keeping the temperature at 50 ℃ for autolysis for 8 hours;
(3) And (3) yeast enzymolysis:
the pH value of the solution after autolysis treatment is adjusted to 5.0, the temperature is 60 ℃, and a compound enzyme preparation containing 2.5 permillage papain, 2.5 permillage alkaline protease, 2.5 permillage neutral protease and 2.5 permillage flavourzyme is added for enzymolysis reaction for 8 hours according to the dry matter of yeast milk.
After the enzymolysis reaction is finished, heating to 75 ℃, preserving heat for 1.5 hours, inactivating enzymes, concentrating and drying to obtain a yeast hydrolysate product E.
The determination of protein content, nucleic acid content, and mannooligosaccharide and beta-glucan content was performed on yeast hydrolysate product E. Wherein the protein content is 48%, the nucleic acid content is 8%, the content of the obtained mannooligosaccharide and beta-glucan is 25%, the content of the mannooligosaccharide is 11.5%, and the content of the beta-glucan is 13.5%.
The main component contents of the yeast hydrolysate products A to E prepared in the examples and comparative examples are summarized in Table 2 below.
TABLE 2 content of principal Components of Yeast hydrolysates in examples and comparative examples
Application example of Yeast hydrolysate to meat chicks
1. Influence of Yeast hydrolysate on the production Performance of meat chicks (death rate; feed to weight ratio; average daily weight gain)
300 chickens (bred in male and female mixed culture) of ROSS 308 meat chicks (purchased from Hubei Dangyang city) are selected and randomly divided into 5 treatment groups, and each treatment group comprises 60 chickens with the test period of 42d.
Each treatment group was fed with a different test diet, and was fed with a base diet, a base diet +1kg/t of the yeast hydrolysate a obtained in example one, a base diet +5kg/t of the yeast hydrolysate D obtained in comparative example one, and a base diet +5kg/t of the yeast hydrolysate E obtained in comparative example two, respectively. The basic ration formula and the nutrition level are shown in table 3, the experimental broiler is free to eat and drink water, the temperature in the range is regulated and controlled by adopting a heat preservation lamp, and the raising management of the broiler and the environmental control in the range are carried out according to the raising management standard of the broiler.
Table 3 basal diet formulation and nutrient levels
| Raw materials | 1-21d | 22-42d | Project | 1-21d | 22-42d |
| Corn | 54.20 | 60.00 | ME(Kcal/kg) | 2964.35 | 3131.27 |
| Soybean meal | 33.00 | 24.50 | Crude protein | 20.99 | 17.98 |
| Secondary powder | 2.00 | 2.50 | Lysine | 1.34 | 1.03 |
| Cottonseed meal | 2.50 | 2.50 | Methionine | 0.52 | 0.41 |
| Rapeseed meal | 1.00 | 2.00 | Methionine+cystine | 0.85 | 0.71 |
| Soybean oil | 3.61 | 5.41 | Calcium | 0.89 | 0.76 |
| Dibasic calcium phosphate | 1.70 | 1.40 | Available phosphorus | 0.45 | 0.39 |
| Stone powder | 1.00 | 0.90 | |||
| Salt | 0.30 | 0.30 | |||
| 99% methionine | 0.22 | 0.15 | |||
| 98.5% lysine | 0.32 | 0.19 | |||
| Choline chloride | 0.10 | 0.10 | |||
| Premix compound | 0.05 | 0.05 |
The measurement indexes are as follows:
rate of death: ratio of dead or eliminated broiler chickens.
Ratio of weight to weight (F/G): the weight ratio and the whole-period weight ratio of each stage are calculated according to the weight gain, death and elimination of the surviving broiler chickens at a certain stage.
Average daily gain: initial body weight was recorded before entry at 8 am at 21, 42 days old: 00, starting to weigh the whole group of test broilers (empty stomach, stop feeding for 12 hours), and calculating the average daily gain of each chicken in each stage.
The results of the effect of the yeast hydrolysate on the death rate and the feed weight ratio of the meat chicks are shown in Table 4 below, and the results of the effect of the yeast hydrolysate on the average weight of the meat chicks are shown in Table 5 below.
TABLE 4 influence of Yeast hydrolysates on the death rate and feed/weight ratio of meat chicks
The same column data shoulder indicates that the difference is not significant (P > 0.05) with the same letter or no letter, and the different lower case letters indicate that the difference is significant (P < 0.05). The table below is the same.
TABLE 5 Effect of Yeast hydrolysates on average body weight of meat chicks
The results show that: the yeast hydrolysate A prepared in the embodiment I is added into daily ration by 1 and 5kg/t, the average weight of 21 days of the ROSS 308 chicks is respectively increased by 1.25 percent and 2.77 percent, the average weight of 42 days of the chicks is respectively increased by 2.74 percent and 3.49 percent, the feed-to-weight ratio of 1 to 21 days of the chicks is respectively reduced by 0.06 percent and the feed-to-weight ratio of 0.10,1 to 42 days of the chicks is respectively reduced by 0.09 percent and the death rate of 0.13,1 to 42 days of the chicks is respectively reduced by 6.58 percent and 4.49 percent.
Adding 5kg/t of yeast hydrolysate D obtained in the comparative example I into daily ration, wherein the average weight of 21 days old of ROSS 308 meat chicks is reduced by 0.32%, and the average weight of 42 days old is reduced by 0.40%; the feed weight ratio of 1-21 days old and 1-42 days old is respectively reduced by 0.03 and 0.01, and the death rate is respectively reduced by-0.31 and 2.84 percent.
Adding 5kg/t yeast hydrolysate E obtained in comparative example II into ration, and increasing average weight of 21-day-old and 42-day-old ROSS 308 broiler chicks by 0.32% and 0.18%, respectively; the feed weight ratio of 1-21 days old and 1-42 days old is reduced by 0.02 and-0.01 respectively, and the death rate is reduced by-0.63 and 2.09 percent respectively.
From the above experimental results, it can be seen that the meat chicks, after eating the yeast hydrolysate prepared in the examples of the present invention, have a decreased dead-wash rate, a decreased feed-to-weight ratio (F/G), and an increased average daily gain, as compared to meat chicks that only eat the basic diet and the yeast hydrolysate prepared in the comparative examples.
2. Influence of Yeast hydrolysate on intestinal morphology of chicks (fluff height)
The following is the measurement of intestinal morphology of the broiler chicken in the experiment, mainly measuring the height (μm) of the nap, and the method is as follows:
selecting 21-day-old chicks, weighing, slaughtering, taking out the intestinal tract, separating the duodenum (U-shaped intestinal loop), the jejunum (the distal end of the duodenum to the diverticulum of the yolk sac) and the ileum (the diverticulum of the yolk sac to the joint of the ileum), peeling off the mesentery and connective tissues, cutting off the mesentery and the connective tissues, flushing with physiological saline, sucking the liquid to dryness by filter paper, cutting off the tissue of the duodenum (the position 5cm away from the pylorus), the middle section of the jejunum and the middle section of the ileum respectively, placing into 8% formalin fixing solution for preservation, flushing the chyme in the intestines by ice physiological saline, fixing for 48 hours by 40g/L paraformaldehyde phosphate buffer (0.1 mol/L, pH 7.4), embedding by conventional paraffin, continuously slicing (6 mu m thick), taking 1 piece every 10 pieces, and dyeing by conventional and glycogen and polysaccharide periodic acid-Schiff (PAS).
After HE staining, observing the mucous membrane structure of the small intestine by using a Leica-DFC450.C microscope, taking non-adjacent 5 sections of small intestine, taking a picture of a straight position, analyzing the picture by using an Image-Pro Plus 5.0 Image analysis system, and respectively taking 5 longest villus heights and the deepest villus for measurement. The effect of yeast hydrolysate on the intestinal morphology of broiler chickens was determined as follows in table 6.
TABLE 6 Effect of Yeast hydrolysates on intestinal morphology of broiler chickens
The results show that: 1kg/t yeast hydrolysate A is added into the daily ration, the intestinal villus height of the duodenum of the ROSS 308 meat chick is respectively increased by 6.96 percent and 13.48 percent, the intestinal villus height of the jejunum is respectively increased by 19.94 percent and 26.93 percent, and the intestinal villus height of the duodenum is respectively increased by 8.78 percent and 12.66 percent.
The yeast hydrolysate D obtained in the first 5kg/t ratio and the yeast hydrolysate E obtained in the second comparative example are added into the daily ration, the intestinal villus height of the duodenum of the ROSS 308 meat chick is respectively increased by 2.68 percent and 4.10 percent, the intestinal villus height of the jejunum is respectively increased by 8.04 percent and 4.88 percent, the intestinal villus height of the duodenum is respectively increased by 6.79 percent and 2.91 percent, and the difference is not obvious.
From the above experimental results, it can be seen that the meat chicks have a higher percentage of intestinal villus height improvement after eating the yeast hydrolysate prepared in the examples of the present invention than the meat chicks eating the yeast hydrolysate prepared in the comparative examples.
3. Influence of Yeast hydrolysate on the infectious bronchitis Positive Rate of meat chicks and the antibody titer against avian influenza
The determination method of the infectious bronchitis positive rate of the broiler chickens and the antibody titer of the avian influenza is as follows:
the chickens are inoculated with newcastle disease vaccine at 1 day of age, and are inoculated with newcastle disease-infectious bronchitis bivalent vaccine and newcastle disease-avian influenza bivalent vaccine at 8 days of age. Then, after 30 days old and 40 days old, 10 feathers are randomly extracted from each repetition, vein blood is collected under the wing, blood is separated and prepared into serum by centrifugation at 3000r/min for 10min, the serum is preserved at-20 ℃, hemagglutination and hemagglutination inhibition tests are carried out, and the positive rate/% > of infectious bronchitis and avian influenza H are measured 5 Antibody titers/log 2 。
The results of the detection of the effect of yeast hydrolysate on the positive rate of infectious bronchitis in broiler chickens and the antibody titer against avian influenza are shown in Table 7 below.
TABLE 7 Effect of Yeast hydrolysate on the positive Rate of infectious bronchitis in broilers, avian influenza antibody titre
The results show that: 1kg/t yeast hydrolysate A is added into daily ration, the infectious bronchitis positive rate of 30 days old of meat chicks is respectively reduced by 33.33 percent and 26.67 percent, the infectious bronchitis positive rate of 40 days old of meat chicks is respectively reduced by 22.50 percent and 18.17 percent, and the avian influenza H of 30 days old of meat chicks 5 The antibody titer is respectively increased by 0.36 log and 0.69 log 2 The titer of the avian influenza H5 antibody of the meat chicks at 40 days of age is respectively improved by 0.57 log and 1.05 log 2 。
The yeast hydrolysate D obtained in the first comparative example of 5kg/t and the yeast hydrolysate E obtained in the second comparative example of 5kg/t are added into daily ration, the positive rate of 30-day-old infectious bronchitis of meat chicks is respectively reduced by 31.62 percent and 24.54 percent, the positive rate of 40-day-old infectious bronchitis of meat chicks is respectively reduced by 20.32 percent and 21.58 percent, and the positive rate of 30-day-old avian influenza H of meat chicks is respectively reduced 5 The antibody titer is respectively increased by 0.62 log and 0.52 log 2 The titer of the avian influenza H5 antibody of the meat chicks at 40 days old is respectively improved by 0.78 log and 0.83 log 2 。
Application example II application of yeast hydrolysate to egg chicks
1. Effect of Yeast hydrolysate on growth and development of egg chicks
The following is an experiment of the influence of the yeast hydrolysate on the production performance of the egg chicks:
360 commercial egg-substituted chicks (purchased from Hubei Dang yang city chicken farm) were selected and randomly divided into 6 treatment groups of 50 egg chicks each for the test period 42d.
Each treatment group was fed with different test diets, and was fed with a base diet, a base diet +5kg/t yeast hydrolysate B prepared in example two, a base diet +10kg/t yeast hydrolysate B prepared in example two, a base diet +20kg/t yeast hydrolysate B prepared in example two, a base diet +5kg/t yeast hydrolysate D prepared in comparative example one, and a base diet +5kg/t yeast hydrolysate E prepared in comparative example two, respectively. The basic ration formula and the nutrition level are shown in table 8, the test egg chicks eat and drink water freely, the temperature in the circle is regulated and controlled by adopting a heat preservation lamp for heating, and the raising management of the laying hens and the environmental control in the house are carried out according to the raising management standard of the laying hens.
Table 8 basic ration formula and nutrient level
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Measuring the index:
average daily gain: initial body weight was recorded before entry at 8 am at 21, 42 days old: 00, the whole group of test egg chicks is weighed (empty stomach, stopping feeding for 12 hours), the average daily gain of each chicken in each stage is calculated, and the detection result is shown in Table 9.
TABLE 9 Effect of Yeast hydrolysates on the growth and development of egg chicks
The results of the growth rate showed that: the average weight of 21 days of the egg chicks is increased by 4.04%, 4.61% and 1.89% respectively, and the average weight of 42 days of the egg chicks is increased by 6.90%, 6.06% and 5.49% respectively by adding 5, 10 and 20kg/t yeast hydrolysate to the daily diet.
The average weight of 21 days old of the egg chicks is reduced by 0.64 percent instead, and the average weight of 42 days old is increased by 1.06 percent by adding 5kg/t of the yeast hydrolysate D obtained in the comparative example I to the daily diet; the average body weight of 21-day and 42-day old chicks was increased by 1.81% and 4.41% respectively by adding 5kg/t of yeast hydrolysate E obtained in comparative example II to the diet.
2. Effect of Yeast hydrolysate on the development of the tibia of an egg chick
The method for measuring the length of the tibia is as follows: at 21 days of age, 10 egg chicks were randomly picked for each treatment and the "tibia length" was measured with vernier calipers: the measurement results are shown in Table 10 below, based on the length (mm) from the upper joint end of the hairless leg to the second toe (little phalanx).
TABLE 10 Effect of Yeast hydrolysates on the tibial development of egg chicks
The results of tibial development indicate that: the average tibia length of 21 days old of the egg chicks is respectively increased by 5.38%, 5.96% and 2.69% by adding 5, 10 and 20kg/t yeast hydrolysate into daily ration.
The average tibia length of 21 days old of the egg chicks is reduced by 1.54% instead by adding the yeast hydrolysate D obtained in the ratio of 5kg/t into the daily ration; the average tibia length of 21 days old of the egg chicks was increased by 0.77% by adding 5kg/t of the yeast hydrolysate E obtained in comparative example II to the diet.
3. Effect of Yeast hydrolysate on antibody titres of newcastle vaccine for egg chicks
Determination of newcastle disease antibody titer of chicks:
the egg chicks were vaccinated with newcastle disease vaccine at 1 day old and with newcastle disease-infectious bronchitis bivalent vaccine and newcastle disease-avian influenza bivalent vaccine at 8 days old. Thereafter, 21 days old, 42 days old, each repetition followedExtracting 10 feather by a machine, collecting blood from subwinged veins, centrifuging at 3000r/min for 10min, separating to obtain serum, preserving at-20deg.C, performing hemagglutination and hemagglutination inhibition test, and determining newcastle disease antibody titer/log 2 The results of the detection of the effect of yeast hydrolysate on newcastle disease antibody titer of chicks are shown in table 11 below.
TABLE 11 Effect of Yeast hydrolysate on Newcastle disease antibody titres in egg-laying chicks
The results of blood immunization showed that: adding yeast hydrolysate 5, 10, 20kg/t into daily ration, and increasing newcastle disease antibody titer of 21 days old of egg chick by 0.44, 0.55, 0.89 log 2 The newcastle disease antibody titer of 42 days old of the egg-laying chicks is respectively improved by 0.33 log, 0.65 log and 0.79 log 2 。
The newcastle disease antibody titer of 21 days old of the egg chicks is reduced by 0.21 log instead by adding the yeast hydrolysate D obtained in the ratio of 5kg/t into the ration 2 While the newcastle disease antibody titer of 42 days old of the egg-laying chicks is improved by 0.08 log 2 The method comprises the steps of carrying out a first treatment on the surface of the The addition of 5kg/t yeast hydrolysate E obtained in comparative example II to diet improves newcastle disease antibody titer of 21-day-old and 42-day-old egg chicks by 0.15 log and 0.3 log, respectively 2 。
In summary, aiming at the problems existing in the current chick breeding, the feeding yeast hydrolysate provided by the invention not only can provide protein, nucleic acid, vitamins and other nutritional ingredients for the chicks, but also can improve intestinal development, increase the chick growth speed, improve the chick immune function and protect the health of young birds.
The above description is not intended to limit the invention in any way, but is intended to cover all modifications, equivalents, and alternatives falling within the spirit and principles of the invention.
Claims (32)
1. A yeast hydrolysate, characterized by comprising the following components in percentage by mass: the total amount of the mannooligosaccharide and the beta-glucan is 31.5-50%, the nucleic acid is 5-7%, and the crude protein is 35-45%; the ratio of the content of the mannooligosaccharide to the content of the beta-glucan is 1:4-4:1;
wherein the yeast hydrolysate is prepared by a preparation method comprising the steps of:
(1) And (3) fermenting and culturing yeast: fermenting and culturing a yeast strain to obtain yeast milk;
(2) Autolysis of yeast: autolysis is carried out on the yeast milk obtained in the step (1);
(3) And (3) yeast enzymolysis: adding a complex enzyme preparation into the product obtained in the step (2), and carrying out enzymolysis to obtain yeast hydrolysate;
the complex enzyme preparation is papain, alkaline protease, neutral protease and flavourzyme;
wherein the fermentation temperature in the step (1) is 25-35 ℃, the fermentation pH is 5.0-7.0, and the fermentation time is 15-18 hours;
the yeast strain is Saccharomyces cerevisiae FX-2 #Saccharomyces cerevisiae FX-2), accession number CCTCC NO: m2016418.
2. The yeast hydrolysate of claim 1, wherein the ratio of the mannooligosaccharide content to the beta glucan content is 1:1.2.
3. The method for producing a yeast hydrolysate according to claim 1 or 2, comprising the steps of:
(1) And (3) fermenting and culturing yeast: fermenting and culturing a yeast strain to obtain yeast milk;
(2) Autolysis of yeast: autolysis is carried out on the yeast milk obtained in the step (1);
(3) And (3) yeast enzymolysis: adding a complex enzyme preparation into the product obtained in the step (2), and carrying out enzymolysis to obtain yeast hydrolysate;
the complex enzyme preparation is papain, alkaline protease, neutral protease and flavourzyme;
wherein the fermentation temperature in the step (1) is 25-35 ℃, the fermentation pH is 5.0-7.0, and the fermentation time is 15-18 hours;
the yeast strain is Saccharomyces cerevisiae FX-2 #Saccharomyces cerevisiae FX-2), accession number CCTCC NO: m2016418.
4. The method according to claim 3, wherein the yeast in the step (1) is fermented in a medium containing a carbon source, a nitrogen source and a phosphorus source.
5. The preparation method according to claim 4, wherein the carbon source is added in an amount of 80% -90% of the culture medium, the nitrogen source is added in an amount of 10% -15% of the culture medium, and the phosphorus source is added in an amount of 1% -5% of the culture medium.
6. The production method according to claim 5, wherein the carbon source is added in an amount of 85% to the medium, the nitrogen source is added in an amount of 12% to the medium, and the phosphorus source is added in an amount of 3% to the medium.
7. The process according to claim 4, wherein the carbon source is molasses and/or starch-hydrolyzing sugar, the nitrogen source is ammonia water and/or ammonium sulfate, and the phosphorus source is phosphoric acid.
8. The production method according to claim 7, wherein the carbon source is molasses and the nitrogen source is ammonia water.
9. The preparation method according to any one of claims 3 to 8, wherein in the step (2), 1 to 5%o of citric acid is further added in percentage by mass.
10. The preparation method according to any one of claims 3 to 8, further comprising a process of heat-beating the yeast milk obtained in step (1) at 85 to 95 ℃ after step (1) and before step (2).
11. The preparation method according to claim 9, wherein the yeast milk obtained in the step (1) is subjected to heat shock at 85-95 ℃ after the step (1) and before the step (2).
12. The preparation method according to any one of claims 3 to 8, wherein in the step (3), 4 to 20% by mass of the complex enzyme preparation is added.
13. The preparation method of claim 12, wherein the compound enzyme preparation contains papain 1-5%o, alkaline protease 1-5%o, neutral protease 1-5%o, and flavourzyme 1-5%o.
14. The preparation method according to claim 9, wherein in the step (3), 4-20% by mass of the complex enzyme preparation is added.
15. The preparation method according to claim 10, wherein in the step (3), 4-20% by mass of the complex enzyme preparation is added.
16. The preparation method according to any one of claims 3 to 8, wherein the time for the enzymolysis in the step (3) is 6 to 12 hours.
17. The preparation method of claim 16, wherein the enzymolysis time in the step (3) is 10-12 hours.
18. The preparation method according to claim 9, wherein the time of the enzymolysis in the step (3) is 6-12 hours.
19. The preparation method of claim 10, wherein the enzymolysis time in the step (3) is 6-12 hours.
20. The preparation method of claim 12, wherein the enzymolysis time in the step (3) is 6-12 hours.
21. The process according to any one of claims 3 to 8, further comprising drying the product obtained in step (3) at 70 to 80 ℃ for 1 to 2 hours.
22. The preparation method according to claim 9, further comprising a process of drying the product obtained in step (3) at 70-80 ℃ for 1-2 hours.
23. The preparation method according to claim 10, further comprising a process of drying the product obtained in step (3) at 70-80 ℃ for 1-2 hours.
24. The process according to claim 12, further comprising drying the product obtained in step (3) at 70 to 80℃for 1 to 2 hours.
25. The process according to claim 16, further comprising drying the product obtained in step (3) at 70 to 80℃for 1 to 2 hours.
26. A feed comprising the yeast hydrolysate of claim 1 or 2.
27. The feed of claim 26, wherein the yeast hydrolysate is added to the feed in an amount of 1-50kg/t.
28. The feed according to claim 26 or 27, wherein the yeast hydrolysate is added to the feed in an amount of 1-20kg/t when fed to meat chickens; when the chicks are fed with the feed, the addition amount of the yeast hydrolysate in the feed is 5-50kg/t.
29. The feed of claim 28, wherein the yeast hydrolysate is added to the feed in an amount of 5kg/t when fed to broiler chicks; when the chicks are fed with the feed, the addition amount of the yeast hydrolysate in the feed is 5-10kg/t.
30. Use of the yeast hydrolysate according to claim 1 or 2 or the yeast hydrolysate prepared by the preparation method according to any one of claims 3-25 in the field of feed.
31. The use according to claim 30, wherein the use is in the field of chicken feed.
32. The use according to claim 30, wherein the use is in the field of chicken feed.
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