CN110669757A - Production method of zinc-rich yeast with high zinc methionine content - Google Patents

Production method of zinc-rich yeast with high zinc methionine content Download PDF

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CN110669757A
CN110669757A CN201910960717.3A CN201910960717A CN110669757A CN 110669757 A CN110669757 A CN 110669757A CN 201910960717 A CN201910960717 A CN 201910960717A CN 110669757 A CN110669757 A CN 110669757A
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zinc
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rich yeast
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朱振华
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HUZHOU GUANGBO BIOTECHNOLOGY Co Ltd
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HUZHOU GUANGBO BIOTECHNOLOGY Co Ltd
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    • 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
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    • 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

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Abstract

The invention discloses a method for producing zinc-rich yeast with high zinc methionine content, which comprises the steps of inoculating a saccharomyces cerevisiae strain into a zinc-containing glucose culture medium to culture and screen out zinc yeast strains, then adopting a culture medium containing chlorella antibacterial peptide powder and a plurality of trace elements to culture the strains, and adding zinc sulfate solution in a flowing manner until fermentation is finished, wherein the zinc-rich yeast finally obtained has the zinc methionine content which can be increased to more than 75 percent.

Description

Production method of zinc-rich yeast with high zinc methionine content
Technical Field
The invention relates to the technical field of yeast, in particular to a production method of zinc-rich yeast with high zinc methionine content.
Background
Zinc is a necessary trace element for animal body and is called "vital element". When the human body lacks zinc, growth retardation can occur; dysplasia in the juvenile period, dysgeusia, slow wound healing, and the like; pregnant women lack zinc and even have fetal deformity. The Chinese is wide in source, the composition of diet is mainly cereal, zinc deficiency is caused by insufficient intake of zinc in food or too much intake of fiber in diet to influence the absorption of zinc, and the zinc deficiency condition of children is common. The daily zinc intake of adults recommended in China is 10-15mg, and according to the analysis of the Chinese preventive medicine academy of sciences on the zinc content in hair of 19 provincial and urban children such as Beijing, about 60 percent of the children are lower than the normal value. The zinc deficiency is serious in the pathological conditions of zinc deficiency, such as poor growth and development of children, dwarfism, dementia and the like. The zinc content in natural food is generally low, and the zinc content in the natural food is not enough to meet the normal requirement of human body.
At present, inorganic zinc such as high-dosage zinc oxide, zinc sulfate and the like is mostly adopted as a zinc nutrition additive in China, but the inorganic zinc has low biological value and large addition amount and is easy to cause serious environmental pollution; the application of the amino acid complex zinc salt (such as zinc methionine, zinc gluconate and the like) with good stability and higher biological value is limited due to the complex production process and higher price. The zinc-rich yeast can not only meet the requirement of animal organisms on zinc, but also has rich nutrient components in the yeast and can play a synergistic role.
The Chinese invention patent CN107119038A discloses a method for producing zinc-rich yeast products, which can produce zinc-rich yeast with zinc content of 1000-5000mg per kilogram of yeast, but the zinc content of methionine in the zinc-rich yeast is less than 30%, which is not beneficial to animal absorption, so that the method for producing zinc-rich yeast with high zinc content of methionine needs to be researched.
Disclosure of Invention
Based on the technical problems in the background art, the invention provides a production method of zinc-rich yeast with high zinc methionine content.
The technical scheme of the invention is as follows:
a production method of zinc-rich yeast with high methionine zinc content comprises the following steps:
A. preparing yeast strains with strong organic zinc conversion capacity: inoculating Saccharomyces cerevisiae strain to zinc-containing glucose culture medium A, culturing, and screening out strain with strong zinc resistance as mutagenesis strain; inoculating yeast strains with strong organic zinc conversion capability into a mutagenesis culture medium for mutagenesis culture for 20-45 h; separating and screening the mutagenized strains, inoculating the strains to a zinc-containing glucose culture medium B for culture, and screening out strains with strong organic zinc transformation capacity, namely the high-transformation zinc-rich yeast strains;
B. and (3) strain culture: putting the culture medium into a seeding tank, sterilizing, and inoculating a high-transformation zinc-rich yeast strain into the culture medium in the seeding tank for culture; the temperature of the seeding tank is 33 +/-3 ℃, the initial pH4.0-4.8, and the culture time is 16 +/-2 hours;
C. and (3) amplification culture: transferring the strain to a fermentation tank for enlarged culture; the temperature of the fermentation tank is 30 +/-2 ℃, the pH value is 5.2-5.8 (the pH value is adjusted by sodium hydroxide), when the wet weight of the thalli reaches 2.1-2.3g/10ml, zinc sulfate solution is fed-batch until the fermentation is finished, and the culture time is 36 +/-2 hours;
D. inactivation: inactivating at 75-77 deg.C for 25-35 min;
E. spray drying: the inlet temperature is 205 ℃ and 230 ℃, the outlet temperature is 85-95 ℃, and the mixture is sieved, mixed and packaged.
Preferably, in the step a, the mutagenesis medium contains a mutagen, and the mutagen in the mutagenesis medium is N-methyl-N-nitro-N-nitrosoguanidine.
Preferably, in the step B, the components of the culture medium are a carbon source, a nitrogen source, inorganic salts, chlorella antibacterial peptide powder and a small amount of trace elements.
Preferably, the nitrogen source is an organic nitrogen source containing amino nitrogen, and more preferably, the organic nitrogen source containing amino nitrogen is a plant protein or an amino acid.
Further preferably, the components of the culture medium consist of the following components in parts by weight: 500 parts of fermentable reducing sugar, 50-70 parts of organic nitrogen source containing amino nitrogen, 6-10 parts of ammonium dihydrogen phosphate, 5-8 parts of magnesium sulfate, 1-3 parts of chlorella antibacterial peptide powder and 0.1-0.3 part of trace elements.
Preferably, the trace elements include: ca2+、Fe2+、Mo2+And Mn2+。
Further preferably, the organic nitrogen source is supplemented every 4 to 8 hours after the zinc sulfate solution is initially fed, and ammonia water is used for controlling the pH value in the process.
The invention has the advantages that: the zinc-rich yeast produced by the method has good stability and high biological value, and the zinc-rich yeast is obtained by inoculating a saccharomyces cerevisiae strain into a zinc-containing glucose culture medium to culture and screen out the zinc-containing yeast strain, then performing strain culture by adopting a culture medium containing chlorella antibacterial peptide powder and various trace elements, and adding zinc sulfate solution in a flowing manner until fermentation is finished.
Detailed Description
Example 1:
a production method of zinc-rich yeast with high methionine zinc content comprises the following steps:
A. preparing yeast strains with strong organic zinc conversion capacity: inoculating Saccharomyces cerevisiae strain to zinc-containing glucose culture medium A, culturing, and screening out strain with strong zinc resistance as mutagenesis strain; inoculating yeast strains with strong organic zinc conversion capability into a mutagenesis culture medium for mutagenesis culture for 35 h; separating and screening the mutagenized strains, inoculating the strains to a zinc-containing glucose culture medium B for culture, and screening out strains with strong organic zinc transformation capacity, namely the high-transformation zinc-rich yeast strains; the zinc content of the glucose culture medium B is higher than that of the glucose culture medium A;
B. and (3) strain culture: putting the culture medium into a seeding tank, sterilizing, and inoculating a high-transformation zinc-rich yeast strain into the culture medium in the seeding tank for culture; the temperature of the seeding tank is 33 +/-3 ℃, the initial pH4.0-4.8, and the culture time is 16 +/-2 hours;
C. and (3) amplification culture: transferring the strain to a fermentation tank for enlarged culture; the temperature of the fermentation tank is 30 +/-2 ℃, the pH value is 5.2-5.8 (the pH value is adjusted by sodium hydroxide), when the wet weight of the thalli reaches 2.15g/10ml, zinc sulfate solution is fed-batch until the fermentation is finished, and the culture time is 36 +/-2 hours;
D. inactivation: inactivating at 75-77 deg.C for 30 min;
E. spray drying: the inlet temperature is 225 ℃, the outlet temperature is 90 ℃, and the mixture is sieved, mixed and packaged.
In the step A, the mutagenesis medium contains a mutagen, and the mutagen in the mutagenesis medium is N-methyl-N-nitro-N-nitrosoguanidine.
In the step B, the components of the culture medium consist of the following components in parts by weight: 500 parts of fermentable reducing sugar, 65 parts of vegetable protein, 8 parts of ammonium dihydrogen phosphate, 6 parts of magnesium sulfate, 1.5 parts of chlorella antibacterial peptide powder and 0.2 part of trace elements.
The chlorella antibacterial peptide powder is freeze-dried powder of chlorella antibacterial peptide concentrated solution.
The trace elements include: ca2+、Fe2+、Mo2+And Mn2+。
And after the zinc sulfate solution is fed in, the organic nitrogen source is replenished once every 6 hours, and the pH value is controlled by ammonia water in the process.
Example 2:
a production method of zinc-rich yeast with high methionine zinc content comprises the following steps:
A. preparing yeast strains with strong organic zinc conversion capacity: inoculating Saccharomyces cerevisiae strain to zinc-containing glucose culture medium A, culturing, and screening out strain with strong zinc resistance as mutagenesis strain; inoculating yeast strains with strong organic zinc conversion capability into a mutagenesis culture medium for mutagenesis culture for 45 h; separating and screening the mutagenized strains, inoculating the strains to a zinc-containing glucose culture medium B for culture, and screening out strains with strong organic zinc transformation capacity, namely the high-transformation zinc-rich yeast strains; the zinc content of the glucose culture medium B is higher than that of the glucose culture medium A;
B. and (3) strain culture: putting the culture medium into a seeding tank, sterilizing, and inoculating a high-transformation zinc-rich yeast strain into the culture medium in the seeding tank for culture; the temperature of the seeding tank is 33 +/-3 ℃, the initial pH4.0-4.8, and the culture time is 16 +/-2 hours;
C. and (3) amplification culture: transferring the strain to a fermentation tank for enlarged culture; the temperature of the fermentation tank is 30 +/-2 ℃, the pH value is 5.2-5.8 (the pH value is adjusted by sodium hydroxide), when the wet weight of the thalli reaches 2.3g/10ml, zinc sulfate solution is fed-batch until the fermentation is finished, and the culture time is 36 +/-2 hours;
D. inactivation: inactivating at 75-77 deg.C for 25 min;
E. spray drying: the inlet temperature is 230 ℃, the outlet temperature is 85 ℃, and the mixture is sieved, mixed and packaged.
In the step B, the components of the culture medium consist of the following components in parts by weight: 500 parts of fermentable reducing sugar, 70 parts of vegetable protein, 6 parts of ammonium dihydrogen phosphate, 8 parts of magnesium sulfate, 1 part of chlorella antibacterial peptide powder and 0.3 part of trace elements.
The chlorella antibacterial peptide powder is freeze-dried powder of chlorella antibacterial peptide concentrated solution.
The trace elements include: ca2+、Fe2+、Mo2+And Mn2+。
And after the zinc sulfate solution is fed in, the organic nitrogen source is replenished every 4 hours, and the pH value is controlled by ammonia water in the process.
Example 3:
a production method of zinc-rich yeast with high methionine zinc content comprises the following steps:
A. preparing yeast strains with strong organic zinc conversion capacity: inoculating Saccharomyces cerevisiae strain to zinc-containing glucose culture medium A, culturing, and screening out strain with strong zinc resistance as mutagenesis strain; inoculating yeast strains with strong organic zinc conversion capability into a mutagenesis culture medium for mutagenesis culture for 20 h; separating and screening the mutagenized strains, inoculating the strains to a zinc-containing glucose culture medium B for culture, and screening out strains with strong organic zinc transformation capacity, namely the high-transformation zinc-rich yeast strains; the zinc content of the glucose culture medium B is higher than that of the glucose culture medium A;
B. and (3) strain culture: putting the culture medium into a seeding tank, sterilizing, and inoculating a high-transformation zinc-rich yeast strain into the culture medium in the seeding tank for culture; the temperature of the seeding tank is 33 +/-3 ℃, the initial pH4.0-4.8, and the culture time is 16 +/-2 hours;
C. and (3) amplification culture: transferring the strain to a fermentation tank for enlarged culture; the temperature of the fermentation tank is 30 +/-2 ℃, the pH value is 5.2-5.8 (the pH value is adjusted by sodium hydroxide), when the wet weight of the thalli reaches 2.1g/10ml, zinc sulfate solution is fed-batch until the fermentation is finished, and the culture time is 36 +/-2 hours;
D. inactivation: inactivating at 75-77 deg.C for 35 min;
E. spray drying: the inlet temperature is 205 ℃, the outlet temperature is 95 ℃, and the mixture is sieved, mixed and packaged.
In the step A, the mutagenesis medium contains a mutagen, and the mutagen in the mutagenesis medium is N-methyl-N-nitro-N-nitrosoguanidine.
In the step B, the components of the culture medium consist of the following components in parts by weight: 500 parts of fermentable reducing sugar, 50 parts of amino acid, 10 parts of ammonium dihydrogen phosphate, 5 parts of magnesium sulfate, 3 parts of chlorella antibacterial peptide powder and 0.1 part of trace elements.
The chlorella antibacterial peptide powder is freeze-dried powder of chlorella antibacterial peptide concentrated solution.
The trace elements include: ca2+、Fe2+、Mo2+And Mn2+。
And after the zinc sulfate solution is fed in, the organic nitrogen source is replenished once every 8 hours, and the pH value is controlled by ammonia water in the process.
Comparative example 1
The chlorella antibacterial peptide powder in the example 1 is removed, and the rest proportion and the preparation method are unchanged.
Comparative example 2
The chlorella antibacterial peptide powder in the example 1 is replaced by the chickpea antibacterial peptide powder, and the rest proportion and the preparation method are unchanged.
Comparative example 3
Ca contained in trace elements in example 12+And Mo2+Removing the components, and keeping the rest proportion and the preparation method unchanged.
The following test results were obtained by testing the zinc-rich yeasts produced in examples 1 to 3 and comparative examples 1 to 3, and the specific results are shown in Table 1.
Table 1: the results of testing the zinc-rich yeasts produced in examples 1 to 3 and comparative examples 1 to 3;
from the above test data, it can be known that the zinc methionine content obtained by the method for producing the zinc-rich yeast with high zinc methionine content of the invention accounts for more than 75% of the total zinc content in the zinc-rich yeast, and the inorganic zinc content is less than 1.5%.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.

Claims (8)

1. A production method of zinc-rich yeast with high methionine zinc content is characterized by comprising the following steps:
A. preparing yeast strains with strong organic zinc conversion capacity: inoculating Saccharomyces cerevisiae strain to zinc-containing glucose culture medium A, culturing, and screening out strain with strong zinc resistance as mutagenesis strain; inoculating yeast strains with strong zinc resistance into a mutagenesis culture medium for mutagenesis culture for 20-45 h; separating and screening the mutagenized strains, inoculating the strains to a zinc-containing glucose culture medium B for culture, and screening out strains with strong organic zinc transformation capacity, namely the high-transformation zinc-rich yeast strains;
B. and (3) strain culture: putting the culture medium into a seeding tank, sterilizing, and inoculating a high-transformation zinc-rich yeast strain into the culture medium in the seeding tank for culture; the temperature of the seeding tank is 33 +/-3 ℃, the initial pH4.0-4.8, and the culture time is 16 +/-2 hours;
C. and (3) amplification culture: transferring the strain to a fermentation tank for enlarged culture; the temperature of the fermentation tank is 30 plus or minus 2 ℃, the pH value is 5.2 to 5.8, when the wet weight of the thalli reaches 2.1 to 2.3g/10ml, zinc sulfate solution starts to be fed-batch until the fermentation is finished, and the culture time is 36 plus or minus 2 hours;
D. inactivation: inactivating at 75-77 deg.C for 25-35 min;
E. spray drying: the inlet temperature is 205 ℃ and 230 ℃, the outlet temperature is 85-95 ℃, and the mixture is sieved, mixed and packaged.
2. The method for producing a zinc-rich yeast with high content of zinc methionine as claimed in claim 1, wherein in the step A, the mutagenizing medium contains a mutagenizing agent, and the mutagenizing agent in the mutagenizing medium is N-methyl-N-nitro-N-nitrosoguanidine.
3. The method for producing a zinc-rich yeast with high content of zinc methionine according to claim 1, wherein in the step B, the culture medium comprises carbon source, nitrogen source, inorganic salt, chlorella antibacterial peptide powder and a small amount of trace elements.
4. The method for producing a zinc-rich yeast having a high content of zinc methionine according to claim 3, wherein the nitrogen source is an organic nitrogen source containing amino nitrogen.
5. The method for producing a zinc-rich yeast with high zinc methionine content according to claim 4, wherein the organic nitrogen source containing amino nitrogen is a plant protein or an amino acid.
6. The method for producing zinc-rich yeast with high content of zinc methionine as claimed in claim 4, wherein the culture medium comprises the following components in parts by weight: 500 parts of fermentable reducing sugar, 50-70 parts of organic nitrogen source containing amino nitrogen, 6-10 parts of ammonium dihydrogen phosphate, 5-8 parts of magnesium sulfate, 1-3 parts of chlorella antibacterial peptide powder and 0.1-0.3 part of trace elements.
7. The method for producing a zinc-rich yeast with high content of zinc methionine as claimed in claim 3 or 4, wherein the trace elements comprise: ca2+、Fe2+、Mo2+And Mn2+。
8. The method for producing a zinc-rich yeast with high zinc methionine content as claimed in claim 1, wherein the organic nitrogen source is supplemented every 4-8 hours after the start of the zinc sulfate solution feeding, and ammonia water is used to control the pH value during the process.
CN201910960717.3A 2019-10-10 2019-10-10 Production method of zinc-rich yeast with high zinc methionine content Withdrawn CN110669757A (en)

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