CN107090483A - Application of the brewer's yeast in glutathione biosynthesis - Google Patents
Application of the brewer's yeast in glutathione biosynthesis Download PDFInfo
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- CN107090483A CN107090483A CN201710271872.5A CN201710271872A CN107090483A CN 107090483 A CN107090483 A CN 107090483A CN 201710271872 A CN201710271872 A CN 201710271872A CN 107090483 A CN107090483 A CN 107090483A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
- C07K5/0215—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing natural amino acids, forming a peptide bond via their side chain functional group, e.g. epsilon-Lys, gamma-Glu
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
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Abstract
The invention discloses application of the brewer's yeast in glutathione biosynthesis, the brewer's yeast was preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on December 26th, 2016, deposit number is CGMCC No.13500, and preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.The present invention extends the application of brewer's yeast, with extensive industrial application value.
Description
Technical field
The present invention relates to application of the brewer's yeast in glutathione biosynthesis, it is in particular to deposit number
Application of the CGMCC No .13500 brewer's yeast in glutathione biosynthesis, belongs to biosynthesis technology field.
Background technology
Glutathione (glutathione, r-glutamyl cysteingl+glycine, GSH) is a kind of containing γ-acyl
The tripeptides of amine key and sulfydryl, is made up of glutamic acid, cysteine and glycine.It is present in each cell of almost body.Paddy
The sweet peptide of Guang can assist in keeping the function of normal immune system, and with antioxidation and integrate detoxication, cysteine
On sulfydryl be its active group（Therefore often it is abbreviated as G-SH）, easily and some drugses（Such as paracetamol）, toxin（Such as free radical,
Iodoacetic acid, mustard gas, the heavy metal such as lead, mercury, arsenic）Deng combination, and with integration detoxication.Glutathione has wide spectrum removing toxic substances
Effect, cannot be only used for medicine, more can as functional food base-material, in the work(such as anti-aging, strengthen immunity, antitumor
Can property food extensive use.
Glutathione（Glutathione, GSH）It is to be combined by glutamic acid, cysteine and glycine, contains sulfydryl
Tripeptides, with antioxidation and integration detoxication.Sulfydryl on cysteine is glutathione active group（Therefore gluathione
Peptide is often abbreviated as G-SH）, easily and some drugses（Such as paracetamol）, toxin（Such as free radical, iodoacetic acid, mustard gas, lead, mercury, arsenic
Deng heavy metal）Deng combination, and with integration detoxication.Therefore glutathione（Glutathione especially in liver cell）It can participate in
Biotransformation, so that the poisonous substance being harmful in body is converted into harmless material, is excreted external.Glutathione can also be helped
Help the function of keeping normal immune system.
Glutathione has reduced form（G-SH）And oxidized form（G-S-S-G）Two kinds of forms, in physiological conditions with reduced form
Glutathione accounts for the overwhelming majority.Change between glutathione reductase catalysis amphitypy.The coenzyme of the enzyme is pentose phosphate shunt generation
The NADPH of offer is provided.
GSH production method mainly has chemical synthesis, solvent extraction, enzyme transforming process and fermentation method.Wherein, fermentation method
It is to utilize cheap carbohydrate, proteinaceous materials, the side of GSH synthesis is carried out by the metabolic pathway of material in microbial cell
Method.The method has the advantages that strain is easy to culture, raw material sources are convenient inexpensively, cost is low, transformation efficiency is high, throughput rate is fast,
Current production GSH main method is turned into.Foreign countries just have been achieved with GSH industrialized production early in 1980s, I
State does not obtain substantive breakthroughs yet due to the industrialized production for the current GSH that starts late.
An Accharomyces cerevisiae, the wine brewing are disclosed in the earlier application CN201710041517.9 of present inventor
Yeast was preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number on December 26th, 2016
For CGMCC No .13500, preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.Follow-up in application inventor is ground
In studying carefully, have now surprisingly been found that the saccharomyces cerevisiae can be also used for the biosynthesis of glutathione, and then complete the present invention.
The content of the invention
The technical problem to be solved in the present invention is to provide a kind of glutathione biological synthesis method.In order to realize above-mentioned mesh
, the present invention uses following technical proposals：
One aspect of the present invention is related to application of the brewer's yeast in glutathione biosynthesis, and the brewer's yeast was in 2016 12
It is preserved within 26th China Committee for Culture Collection of Microorganisms's common micro-organisms center the moon, deposit number is CGMCC No
.13500, preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.
Used brewer's yeast is recorded in earlier application CN201710041517.9 in the present invention, the earlier application
Full text be incorporated into description of the invention and exist as part of the specification, applicant retain be based on the earlier application
Change the right of the present invention.
Application of the brewer's yeast in glutathione biosynthesis, the brewer's yeast was preserved on December 26th, 2016
China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No .13500, preservation address
For Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.
In a preferred embodiment of the present invention, intracellular glutathione content is 95-100mg/ afterwards by fermentation
g。
In a preferred embodiment of the present invention, the culture medium for biosynthesis includes fermentation medium, mixing
Vitamin solution and precusor amino acids solution, it is preferred that described precusor amino acids solution includes sodium glutamate, the Guang ammonia of L- half
Acid hydrochloride and glycine.
The present invention extends the application of brewer's yeast, with extensive industrial application value.
Microorganism Deposit Information
Saccharomyces cerevisiae (Saccharomyces cerevisiae), the saccharomyces cerevisiae was preserved in China on December 26th, 2016
Microbiological Culture Collection administration committee common micro-organisms center, deposit number is CGMCC No .13500, and preservation address is north
The institute 3 of Jing Shi Chaoyang Districts North Star West Road 1.
Embodiment
The technology contents of the present invention are described in detail with reference to specific embodiment, for the ease of entering with prior art
Row compares, and method of the present invention with reference to disclosed in Chinese patent application CN105505801A determines beer ferment involved in the present invention
Female ability in biosynthesis of glutathione.
Embodiment 1：
(1) strain
Brewer's yeast was preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on December 26th, 2016,
Deposit number is CGMCC No .13500.
(2) culture medium
Slant medium (g/L)：Glucose 20, peptone 20, yeast extract 10, the .5 of pH 4,20,121 DEG C of sterilizings of agar
20min；
Seed culture medium (g/L)：Glucose 20, peptone 20, yeast extract 10, NaCl 5, the .0 of .5 of pH 4~6,121 DEG C go out
Bacterium 20min；
Fermentation medium (g/L)：Glucose 50, yeast extract 10, urea 4, (NH4)2SO44, (NH4 )2HPO4 10, KH2PO4
2 .5, Na2HPO40 .5, MgSO4﹒ 7H2O 1, CaCl21, pH 4 .5~6 .0,121 DEG C of sterilizing 20min；
Mixed vitamin solution (g/L)：Vitamin B1 1, vitamin B2 1, vitamin B3 1, adenine phosphate 1, vitamin
B6 3, calcium pantothenate 1；
Precusor amino acids solution (g/L)：Sodium glutamate 40, L-cysteine hydrochloride 40, glycine 10.
(3) actication of culture
Slant medium sterilizing is standby, and strain transfer is activated in test tube slant, 28~32 DEG C of 36~48h of culture.
(4) prepared by seed liquor
Seed culture medium packing (100ml/500ml shaking flasks) sterilizing is standby, takes the strain transfer after activation in seed culture medium
In.28~32 DEG C, 200~220rpm shaking table cultures, 18~20h.
(5) inoculation fermentation culture medium ferments
Fermentation medium packing (100ml/500ml shaking flasks) sterilizing is standby, takes 5mL mixed vitamins solution and 10ml seed liquors
(inoculum concentration 8~10%) is added in fermentation medium, 28~32 DEG C, 200~220rpm shaking table cultures.In shaking table culture 12h,
30% glucose solution 15mL is added during 24h respectively, 15mL precusor amino acids solution, shaking table training are added in shaking table culture 24h
Support and terminate fermentation after 36~42h.
(6) breaking yeast cellule membrane release glutathione
The bacterium solution of certain volume is taken in centrifuge tube, 5000~8000rpm centrifuges 5~10min and collects thalline, brine
After somatic cells 2~3 times, with distillation aqueous suspension, -80 DEG C of refrigerators are put into after shaking up and are freezed rapidly, by the thalline of frost in boiling water
Melt rapidly under the conditions of bath, 5000~8000rpm centrifuges 5~10min after multigelation 2 times, takes supernatant as test sample.
(7) measure of dry cell weight
The bacterium solution of certain volume is taken in centrifuge tube, 5000~8000rpm centrifuges 5~10min and collects thalline, brine
After somatic cells 2~3 times, obtained wet thallus is dried under the conditions of 105 DEG C to constant weight.
(8) measure of intracellular glutathione content
Using iodometric determination supernatant Glutathione peptide content, intracellular gluathione is calculated in conjunction with sporoderm-broken rate and dry cell weight
The content of peptide.
(9) result
Fermentation time：36-42 hours；Dry cell weight：10-15g/L；Intracellular glutathione content：95-100mg/g.
Described above is the preferred embodiments of the present invention, it is noted that come for those skilled in the art
Say, on the premise of principle of the present invention is not departed from, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.For the technical staff of general domain, on the premise of without departing substantially from true spirit
To any obvious change that it is done, it will all constitute to infringement of patent right of the present invention, corresponding law duty will be undertaken
Appoint.
Claims (4)
1. application of the brewer's yeast in glutathione biosynthesis, the brewer's yeast is on December 26th, 2016 is preserved in
State's Microbiological Culture Collection administration committee common micro-organisms center, deposit number is CGMCC No .13500, and preservation address is
Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.
2. the application described in claim 1, intracellular glutathione content is 95-100mg/g afterwards by fermentation.
3. application according to claim 1, it is molten that the culture medium for biosynthesis includes fermentation medium, mixed vitamin
Liquid and precusor amino acids solution, it is preferred that described precusor amino acids solution includes sodium glutamate, L-cysteine hydrochloride
And glycine.
4. using the glutathione of synthesis described in claim 1-3 any one.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710271872.5A CN107090483B (en) | 2017-04-24 | 2017-04-24 | Application of saccharomyces cerevisiae in glutathione biosynthesis |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710271872.5A CN107090483B (en) | 2017-04-24 | 2017-04-24 | Application of saccharomyces cerevisiae in glutathione biosynthesis |
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| Publication Number | Publication Date |
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| CN107090483A true CN107090483A (en) | 2017-08-25 |
| CN107090483B CN107090483B (en) | 2021-03-05 |
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| CN201710271872.5A Active CN107090483B (en) | 2017-04-24 | 2017-04-24 | Application of saccharomyces cerevisiae in glutathione biosynthesis |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6434279A (en) * | 1987-07-29 | 1989-02-03 | Ajinomoto Kk | Production of yeast with high glutathione content |
| CN101054560A (en) * | 2007-04-03 | 2007-10-17 | 中国药科大学 | Biosynthesis method for glutathione by enzyme engineering technique |
| CN102994405A (en) * | 2012-08-14 | 2013-03-27 | 湖北工业大学 | Saccharomyces cerevisiae and application thereof |
| CN105505801A (en) * | 2015-12-25 | 2016-04-20 | 湖北泱盛生物科技有限公司 | Saccharomyces cerevisiae for high yield of glutathione and application of saccharomyces cerevisiae |
-
2017
- 2017-04-24 CN CN201710271872.5A patent/CN107090483B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6434279A (en) * | 1987-07-29 | 1989-02-03 | Ajinomoto Kk | Production of yeast with high glutathione content |
| CN101054560A (en) * | 2007-04-03 | 2007-10-17 | 中国药科大学 | Biosynthesis method for glutathione by enzyme engineering technique |
| CN102994405A (en) * | 2012-08-14 | 2013-03-27 | 湖北工业大学 | Saccharomyces cerevisiae and application thereof |
| CN105505801A (en) * | 2015-12-25 | 2016-04-20 | 湖北泱盛生物科技有限公司 | Saccharomyces cerevisiae for high yield of glutathione and application of saccharomyces cerevisiae |
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| CN107090483B (en) | 2021-03-05 |
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