CN114292763A - Saccharomyces cerevisiae capable of highly producing gamma-aminobutyric acid and soluble beta-glucan and application thereof - Google Patents
Saccharomyces cerevisiae capable of highly producing gamma-aminobutyric acid and soluble beta-glucan and application thereof Download PDFInfo
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Abstract
The invention relates to the technical field of microbial fermentation, in particular to saccharomyces cerevisiae for high yield of gamma-aminobutyric acid and soluble beta-glucan and application thereof. The invention obtains the saccharomyces cerevisiae with high yield of gamma-aminobutyric acid and soluble beta-glucan by screening artificially in the nature and combining the modes of ultraviolet mutagenesis, ARTP mutagenesis and ultraviolet-ARPT combined mutagenesis. The saccharomyces cerevisiae of the invention has been preserved in China center for type culture Collection at 11/12/2021, with the preservation address: the preservation number of Wuhan university in Wuhan City of China is CCTCC NO: M20211406. The saccharomyces cerevisiae of the invention improves the yield of GABA in the fermentation liquor to 7.4g/L and the yield of soluble beta-glucan to 3.5g/L, thus realizing the high-efficiency production of gamma-aminobutyric acid and soluble beta-glucan.
Description
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to saccharomyces cerevisiae for high yield of gamma-aminobutyric acid and soluble beta-glucan and application thereof.
Background
Gamma-aminobutyric acid (GABA), also known as amino-complex acid, is a natural amino acid widely found in animals, plants and microorganisms, obtained by catalytic conversion of Glutamic acid by glutamate decarboxylase (GAD). GABA is a natural anti-aging factor, and has effects of relieving muscle and reducing wrinkle. Currently, GABA is mainly obtained by three ways, namely chemical synthesis, plant extraction and microbial fermentation. The chemical synthesis of the GAB raw material has high cost, relates to an organic solvent, has poor separation effect, great safety threat and larger limitation on the use range, and can not be used in cosmetics, foods and medicines; the plant extraction yield is low and the separation is difficult; in contrast, the microbial fermentation synthesis of GABA is a safer and economically practical method.
Beta-glucan (β -glucan) is a polysaccharide formed by connecting glucose monomers through β -glycosidic bonds and is widely found in fungi, bacteria and plants. Fungi are one of the important sources of beta-glucans, which are composed primarily of beta-1, 3 glycosidic linkages, with a small number of beta-1, 6 glycosidic linkages also being present. One part of the fungus beta-glucan is from the cell wall of the fungus, and the glucan in the cell wall of the fungus is beta-1, 3-1, 6-glucan, wherein 65-90% of the glucan is beta-1, 3-glucan.
Saccharomyces cerevisiae is a safe bacterium widely used in cosmetics, and is favored by people due to the fact that the Saccharomyces cerevisiae contains rich skin care factors. These skin care factors include yeast beta glucan. The yeast beta-glucan is a cell wall structure polysaccharide which takes beta-1, 3 glycosidic bond as a main bond and beta-1, 6 glycosidic bond as an auxiliary bond, wherein about 85 percent of the polysaccharide is insoluble in water. The cell wall of the saccharomyces cerevisiae accounts for about 20% of the dry weight of the cell, the thickness of the cell wall is about 100-200 nm, the cell wall is divided into three layers from outside to inside, and the components of the cell wall are mannose, protein and beta-glucan respectively. Beta-glucans in the cell wall of saccharomyces cerevisiae can be divided into two categories: beta-1, 3 glucan as a main component forming a network structure; beta-1, 6 glucan as a branch to other components.
The soluble yeast beta glucan has good effects of moisturizing, resisting oxidation and repairing cells damaged by ultraviolet irradiation, and is favored in the cosmetic industry. At present, the preparation of soluble yeast beta-glucan mainly takes yeast cell walls as raw materials and adopts acid extraction, alkali extraction, acid-alkali extraction method and enzyme method to prepare the soluble yeast beta-glucan. The methods have low production efficiency, low product purity, large loss of glucan activity and environmental pollution. Therefore, the autonomous secretion of soluble yeast beta-glucan by yeast is the key to solving these problems.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides saccharomyces cerevisiae for high yield of gamma-aminobutyric acid and soluble beta-glucan and application thereof.
The invention provides saccharomyces cerevisiae for high yield of gamma-aminobutyric acid and soluble beta-glucan aiming at the problems that the yield of gamma-aminobutyric acid and soluble beta-glucan prepared by a traditional method in the prior art is low, the pollution is large, the steps are complicated and the market demand is difficult to meet. The invention obtains a saccharomyces cerevisiae capable of highly producing gamma-aminobutyric acid and soluble beta-glucan by using a mode of ultraviolet mutagenesis, ARTP mutagenesis and ultraviolet-ARTP combined mutagenesis.
In order to achieve the purpose, the invention adopts the technical scheme that: providing a saccharomyces cerevisiae strain with high yield of gamma-aminobutyric acid and soluble beta-glucan, wherein the saccharomyces cerevisiae has been preserved in China center for type culture Collection at 11, 12 months in 2021 at the preservation address: the preservation number of Wuhan university in Wuhan City of China is CCTCC NO: M20211406.
The invention also provides a microbial agent, which comprises the saccharomyces cerevisiae.
The invention also provides application of the saccharomyces cerevisiae in preparation of gamma-aminobutyric acid or soluble beta-glucan.
The invention also provides application of the saccharomyces cerevisiae in preparing cosmetics containing gamma-aminobutyric acid or soluble beta-glucan.
The invention also provides application of the saccharomyces cerevisiae in preparing health care products containing gamma-aminobutyric acid or soluble beta-glucan.
The invention also provides a method for simultaneously producing gamma-aminobutyric acid and soluble beta-glucan, wherein the method is to use the saccharomyces cerevisiae for fermentation production.
As a preferred embodiment of the process according to the invention, the fermentation is carried out using a medium comprising: 5-10 g/L yeast extract powder, 15-30 g/L peptone, 10-20 g/L malt extract powder, 10-30 g/L glucose, 10-30 g/L sucrose, 20-40 g/L sodium chloride, 4-10 g/L-glutamic acid and 0.2-0.8 g/L pyridoxal phosphate.
As a preferred embodiment of the method of the present invention, the fermentation conditions are: the fermentation temperature is 25-32 ℃, the pH value in the fermentation process is controlled to be 4.5-6.5, the rotating speed of a fermentation tank is 200-800 rpm, the amount of dissolved oxygen in the fermentation liquid is 20-80%, and the fermentation time is 48-72 h.
The invention has the beneficial effects that:
(1) the invention provides a saccharomyces cerevisiae capable of simultaneously producing gamma-aminobutyric acid and soluble beta-glucan in high yield. The invention adopts the ultraviolet and ARTP mutagenesis method, combines the optimization of fermentation conditions, and utilizes microbial cells to realize the high-efficiency production of the gamma-aminobutyric acid and the soluble beta-glucan.
(2) According to the saccharomyces cerevisiae strain, the yield of GABA in fermentation liquor is improved to 7.4g/L and the yield of soluble beta-glucan is improved to 3.5g/L through culture medium optimization and fermentation condition optimization on a fermentation tank.
(3) After the saccharomyces cerevisiae is cultured under the optimized condition, the antioxidant property of fermentation liquor is improved, namely screened strains are inoculated into an optimized culture medium for fermentation, and after fermentation, the clearance rate of the fermentation liquor with the concentration of 30% on DPPH free radicals is improved to 99.5% from 34.3% of the original strain.
Drawings
FIG. 1 is a graph showing the yield assay corresponding to the primary screening of high GABA-producing and soluble β -glucan strains.
FIG. 2 is a graph showing the yield assay for screening strains with high GABA yield and soluble β -glucan by UV mutagenesis.
FIG. 3 is a graph showing the yield measurement corresponding to the screening of strains highly producing GABA and soluble ss-glucan by ARTP mutagenesis.
FIG. 4 is a graph showing the yield measurement corresponding to screening of a strain highly producing GABA and soluble ss-glucan by ultraviolet-ARTP mutagenesis.
Detailed Description
The technical scheme of the invention is further illustrated by the following specific examples. The examples are given solely for the purpose of illustration and are not intended to limit the scope of the invention. The experimental methods used in the following examples are all conventional methods unless otherwise specified; the materials and reagents used, etc., were all commercially available materials and reagents unless otherwise specified.
1. GABA screening medium (g/L): 10.0 parts of yeast extract powder, 20.0 parts of peptone, 20.0 parts of agar, 20.0 parts of glucose, 0.1 part of bromocresol green, 2 parts of glyoxylic acid and 2 parts of succinic acid.
2. Detecting GABA by a high performance liquid chromatography: using the purchased GABA standard sample as a standard sample, different concentrations of 0.5, 1, 2, 3, 4 and 5g/L were prepared to prepare a standard curve. The GABA yield was calculated from the peak area by gradient elution using a column C18 (5.0X 250mm), mobile phase A (0.5% aqueous sodium acetate) and mobile phase B (0.5% aqueous sodium acetate-methanol), and measuring the absorption peak at a wavelength of ultraviolet 338 nm.
3. Detection of soluble β -glucan by the sulphuric acid phenol method: respectively and precisely absorbing 0.1ml, 0.2 ml, 0.3 ml, 0.4 ml and 0.5ml of glucose standard solution, placing the glucose standard solution in 10ml test tubes with plugs, adding water to 0.5ml in each tube, respectively adding 3ml of sodium tetraborate-sulfuric acid solution in ice water bath, uniformly mixing by using a vortex mixer, heating for 5min in boiling water bath, taking out, immediately cooling to room temperature, adding 0.1ml of 0.1% carbazole solution, shaking up, boiling for 5min, cooling to room temperature, and measuring the absorbance at 530 nm. And (3) performing linear regression by taking the concentration as an abscissa and the absorbance value as an ordinate to prepare a standard curve. The experimental samples were tested in the same manner and the soluble β -glucan yield was calculated by substituting the absorbance values into a standard equation.
Example 1: screening of high-yield GABA (Gamma amino acid butyric acid) saccharomyces cerevisiae strains
20g of soil at a wet place of about 5cm in the orchard is taken, the soil is placed in a 500mL triangular flask, 200mL of sterile phosphate buffer (pH5.0) is added, sodium glutamate with the final concentration of 20g/L is added, a magnetic stirrer is used for stirring uniformly, and then the mixture is placed at 30 ℃ for incubation for 12 hours. Then taking appropriate amount of supernatant to respectively dilute 10-4、10-5And 10-6After doubling, 50. mu.l of the plate was coated with GABA screening plate. After the single colony grows out, a round wet smooth dark green single colony is picked. And then, carrying out streak purification on the selected single colony, screening to obtain 50 strains of GABA-producing strains, and storing for later use.
Example 2: screening of high-yield GABA and soluble beta-glucan strains
50 GABA-producing strains selected in example 1 were inoculated into 250mL triangular flasks each containing 50mL YPD, and cultured on a shaker at 220rpm at 30 ℃ for 48 hours. After fermentation is finished, taking fermentation liquor, centrifuging for 10min at 5000g, collecting supernatant, measuring gamma-aminobutyric acid by using a high performance liquid phase, and detecting the content of soluble beta-glucan by using a phenol sulfate method. As can be seen from the results, 6 strains among the analyzed strains were able to synthesize GABA and soluble ss-glucan at the same time, and the yields thereof are shown in FIG. 1. Wherein the strain S6 has the highest GABA yield reaching 0.98g/L, and the yield of the strain for simultaneously synthesizing the soluble beta-glucan is 0.18 g/L. The GABA yield of the strain S5 is 0.74g/L, the soluble beta-glucan yield is 0.38g/L, and the strain S5 is the strain with the highest soluble beta-glucan yield among 6 strains.
Example 3: identification of high-yield GABA and soluble beta-glucan saccharomyces cerevisiae strains
And identifying the 6 strains which are obtained by screening and simultaneously have high yield of GABA and soluble beta-glucan, using a saccharomyces cerevisiae S288C genome as a template, amplifying an 18S rDNA sequence (shown as SEQ ID NO: 1) of the strain to be detected by using an 18S rDNA universal primer, and sequencing. Compared with an NCBI website, the result shows that the similarity of the strain and the 18S rDNA sequence of the saccharomyces cerevisiae strain is as high as 99.9 percent, and the strain belongs to the saccharomyces cerevisiae through phylogenetic tree clustering analysis.
Example 4: saccharomyces cerevisiae strain for screening high-yield GABA and soluble beta-glucan by ultraviolet mutagenesis
And (3) taking 6 strains of yeast capable of producing GABA and soluble yeast beta-glucan obtained by primary screening, respectively inoculating the strains of yeast into YPD culture media, and culturing the strains of yeast in a shaking table at 30 ℃ and 220rpm for 8-12 h to enter a logarithmic phase. Selecting yeast cultured to logarithmic growth phase for mutagenesis treatment. In a dark room, the ultraviolet lamp is turned on, and the equipment is preheated for 20 min. 10-15 mL of yeast suspension with different dilutions is placed in a sterile empty culture dish. Wearing ultraviolet-proof sterile gloves, opening the cover of the culture dish at a distance of 25cm under a 30W ultraviolet lamp, slightly shaking the bacterial suspension in the dish, and respectively irradiating for different times. 1mL of treated bacteria liquid is taken out of the bacteria liquid and placed on ice to be cooled for 1-2 h, DNA repair in the bacteria body can be inhibited under the low-temperature condition, and mutation efficiency is increased. 200. mu.L of the cooled and placed bacterium solution was applied to YPD medium plates and cultured at 30 ℃ in the dark for 3 days. 500 normal single colonies grow randomly, are inoculated into a 96-hole deep-hole plate filled with 2mL YPD culture medium, are cultured for 72 hours at 30 ℃, are centrifuged, and the supernatant is taken out to detect the yield of GABA and soluble beta-glucan by a high performance liquid chromatography method and a phenol sulfate method respectively. The yield of both products was not improved or even decreased in most strains as measured. Among them, 13 strains showed good results after mutation, and both products showed different improvements, and the results are shown in FIG. 2. Wherein the highest yield of GABA is 2.6g/L (strain uvM13), and the highest yield of soluble beta-glucan is 0.45g/L (strain uvM 9).
Example 5: saccharomyces cerevisiae strain for screening high-yield GABA and soluble beta-glucan by ARTP mutagenesis
And (3) taking 6 strains of yeast capable of producing GABA and soluble yeast beta-glucan obtained by primary screening, respectively inoculating the strains of yeast into YPD culture media, and culturing the strains of yeast in a shaking table at 30 ℃ and 220rpm for 8-12 h to enter a logarithmic phase. Sucking 1mL of bacterial liquid cultured to logarithmic phase in an ultraclean workbench, centrifuging at 6000g for 10min, discarding supernatant, washing for 3 times with sterile physiological saline, sucking 10 mu L of bacterial liquid, placing the bacterial liquid on a slide, sequentially placing the slide in corresponding grooves with sterile tweezers, fixing a centrifugal tube filled with 1mL of YPD liquid culture medium below, and carrying out mutagenesis test. Helium with the purity of 99.999 percent is selected as working gas, the flow rate is 10sL/min, the power supply power is set to be 120W, the operation temperature is 20 ℃, and the processing time is 0, 20, 40, 60, 80, 100, 120 and 140 seconds respectively. After mutagenesis, the centrifuge tube with the slide glass is placed on a vortex machine to be violently shaken for 1min, mutagenized bacteria are eluted, 200 mu L of mutagenized bacteria are coated on a flat plate, and the flat plate is cultured for 72 hours in an incubator at 30 ℃ in the dark. 500 single colonies growing on the plates were randomly picked, inoculated into a 96-well deep-well plate containing 2mL of YPD medium, cultured at 30 ℃ for 72h, centrifuged to take the supernatant for detecting the yield of GABA and soluble beta-glucan, wherein 15 strains have good results after mutation, and the results are shown in FIG. 3. Wherein the highest yield of GABA is 2.9g/L (strain aM13), and the highest yield of soluble beta-glucan is 0.59g/L (strain aM 13).
Example 6: saccharomyces cerevisiae strain for screening high-yield GABA and soluble beta-glucan by ultraviolet-ARTP combined mutagenesis
In a dark room, the ultraviolet mutagenic instrument is started, the equipment is preheated, and 10mL OD is taken600Pouring the yeast suspension of 0.2 into a sterile plate, taking ultraviolet-proof sterile gloves, opening the cover of the culture plate under a 30W ultraviolet lamp at a distance of 25cm, and shaking the plate clockwise and smoothly to enable the yeast suspension in the plate to be uniformly irradiated by ultraviolet rays. After ultraviolet mutagenesis treatment for 1 minute, 10 mu L of irradiated strain suspension is taken and placed on an ARTP mutagenesis instrument slide, the slide is sequentially placed into corresponding grooves by using sterile tweezers, a centrifugal tube filled with 1mL of YPD liquid culture medium is fixed below, and then the mutagenesis test can be started, wherein the mutagenesis time is 100 seconds. After mutagenesis, the centrifuge tube with the slide was placed on a vortex shaker and shaken vigorously for 1 minute to elute the mutagenized bacteria, 200. mu.L of which was spread on YPD solid plates and incubated in an incubator at 30 ℃ for 72 hours in the dark. After single colonies grow out, 500 single colonies are picked and inoculated into a 96-hole deep-hole plate filled with 2mL YPD medium, and fermentation is carried out for 72 hours at 30 ℃ and 220 rpm. After the fermentation is finished, taking out the fermentationCentrifuging the solution at 5000g for 10min, collecting supernatant, and measuring GABA and soluble beta-glucan content. Among them, 10 strains showed good results after mutation, and the results are shown in FIG. 4. Wherein the GABA yield of the optimal strain uvaM10 is 4.9g/L, and the highest yield of the soluble beta-glucan is 1.4 g/L.
The optimal strain uvaM10 of the invention is deposited in China center for type culture Collection on 12/11/2021, address: the preservation number of Wuhan university in Wuhan City of China is CCTCC NO: M20211406.
Example 7: genetic stability analysis of high-yield GABA and soluble beta-glucan saccharomyces cerevisiae
The optimal strain uvaM10 obtained by the mutation is inoculated in a YPD liquid culture medium and cultured for 24h, and then passage is carried out for 20 times in sequence according to the inoculum concentration of 2%, and each passage is cultured for 24 h. The final passage strain is inoculated in YPD culture medium according to the inoculation amount of 10 percent, and cultured at 30 ℃ for 72h to detect the yield of GABA and soluble beta-glucan. The results were consistent with those in example 6, and no yield reduction occurred. The mutant strain is stable in hereditary character and can stably produce GABA and soluble beta-glucan.
Example 8: shake flask culture medium optimization of high-yield GABA and soluble beta-glucan saccharomyces cerevisiae
The above optimal strain uvaM10 was inoculated in the following medium at an inoculum size of 10%: 15g/L of sucrose, 20g/L of glucose, 20g/L of malt extract powder, 20g/L of peptone, 10g/L of yeast extract powder, 10g/L of sodium chloride, 10g/L of L-glutamic acid and 0.2g/L of pyridoxal phosphate. After shaking culture for 72h, centrifuging for 10min at 5000g, taking fermentation supernatant, and measuring GABA and soluble beta-glucan content. The yield of the microzyme GABA in the fermentation supernatant is improved to 5.8g/L, and the yield of the soluble beta-glucan is improved to 2.1 g/L.
Example 9: amplifying and culturing high-yield GABA and soluble beta-glucan saccharomyces cerevisiae in a 10L fermentation tank
The fermentation medium is (g/L): 60 parts of cane sugar, glucose and malt extract powder respectively, 20 parts of yeast extract powder, 10 parts of peptone, 10 parts of sodium chloride, 30 parts of L-glutamic acid and 2 parts of pyridoxal phosphate. The fermentation conditions of the 10L fermentation tank are as follows: the fermentation temperature is 30 ℃, the fermentation pH is 5.0, the initial rotation speed of the fermentation is 200rpm, and the fermentation time is 96 h.
A single colony of uvaM10 was picked and inoculated into a flask containing 200ml YPD medium, and cultured on a shaker at 30 ℃ and 220rpm for 24 hours to obtain a seed solution. Inoculating the cultured seed liquid into a 10L fermentation tank according to the inoculation amount of 10%, adjusting the pH value by ammonia water in the fermentation process, and coupling the dissolved oxygen and the rotating speed to keep the dissolved oxygen at 20-50% when the dissolved oxygen is reduced to below 20%. And after fermenting for 96 hours, centrifuging and taking fermentation liquor to determine the content of GABA and soluble beta-glucan. The yield of the microzyme GABA in the fermentation supernatant is increased to 7.4g/L from 5.8g/L of the shake flask fermentation, and the content of the soluble beta-glucan is increased to 3.5g/L from 2.1g/L of the shake flask fermentation.
Example 10: mutant strain uvaM10 fermentation liquor antioxidant activity analysis
DPPH 1mg was weighed out and dissolved in 20mL of ethanol, and the solution was fully dissolved by shaking. Taking a proper amount of DPPH solution, measuring the light absorption value A at 517nm, adjusting the light absorption value A to be 0.8-1.0 by using ethanol, and then storing in dark for later use. Taking supernatant of fermentation liquor cultured by a shake flask, diluting the supernatant to 30% concentration by using purified water, taking 2mL of the supernatant, putting the supernatant into a centrifuge tube, adding 2mL of the prepared DPPH solution, quickly and uniformly mixing, reacting for 30min in a dark condition, and then measuring the light absorption value As of a corresponding sample at 517 nm; the blank was replaced with water and the absorbance was A0. DPPH radical clearance (%) ((a0-As)/a0) × 100. The clearance rate of the uvaM10 mutant strain fermentation broth supernatant to free radicals is calculated to reach 99.5%.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
SEQUENCE LISTING
<110> whole county (Shanghai) Biotech limited
<120> saccharomyces cerevisiae for high yield of gamma-aminobutyric acid and soluble beta-glucan and application thereof
<130> 2021.12.16
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 822
<212> DNA
<213> Saccharomyces cerevisiae
<400> 1
caagtttgct gggggccccc tacctgattt gaggtcaaac tttaagaaca ttgttcgcct 60
agacgctctc ttcttatcga taacgttcca atacgctcag tataaaaaaa gattagccgc 120
agttggtaaa acctaaaacg accgtacttg cattatacct caagcacgca gagaaacctc 180
tctttggaaa aaaaacatcc aatgaaaagg ccagcaattt caagttaact ccaaagagta 240
tcactcacta ccaaacagaa tgtttgagaa ggaaatgacg ctcaaacagg catgccccct 300
ggaataccaa ggggcgcaat gtgcgttcaa agattcgatg attcacggaa ttctgcaatt 360
cacattacgt atcgcatttc gctgcgttct tcatcgatgc gagaaccaag agatccgttg 420
ttgaaagttt ttaatatttt aaaatttcca gttacgaaaa ttcttgtttt tgacaaaaat 480
ttaatgaata gataaaattg tttgtgtttg ttacctctgg gccccgattg ctcgaatgcc 540
caaagaaaaa gttgcaaaga tatgaaaact ccacagtgtg ttgtattgaa acggttttaa 600
ttgtcctata acaaaagcac agaaatctct caccgtttgg aatagcaaga aagaaactta 660
caagcctagc aagaccgcgc acttaagcgc aggcccggct ggactctcca tctcttgtct 720
tcttgcccag taaaagctct catgctcttg ccaaaacaaa aaaatccatt ttcaaaatta 780
ttaaatttct ttaatgatcc ttccgcaggt tcaccctacg ga 822
Claims (8)
1. The saccharomyces cerevisiae with high yield of gamma-aminobutyric acid and soluble beta-glucan is preserved in the China center for type culture Collection at 11 month and 12 days 2021 at the preservation address: the preservation number of Wuhan university in Wuhan City of China is CCTCC NO: M20211406.
2. A microbial inoculant comprising the saccharomyces cerevisiae of claim 1.
3. Use of the saccharomyces cerevisiae according to claim 1 for the preparation of gamma-aminobutyric acid or soluble beta-glucan.
4. Use of the saccharomyces cerevisiae yeast according to claim 1 for the preparation of cosmetics containing gamma-aminobutyric acid or soluble beta-glucan.
5. The use of the saccharomyces cerevisiae as claimed in claim 1 for the preparation of a health product containing gamma-aminobutyric acid or soluble beta-glucan.
6. A method for simultaneously producing gamma-aminobutyric acid and soluble beta-glucan, wherein the method is a fermentation production by using the Saccharomyces cerevisiae according to claim 1.
7. The method of claim 6, wherein the fermentation medium comprises: 5-10 g/L yeast extract powder, 15-30 g/L peptone, 10-20 g/L malt extract powder, 10-30 g/L glucose, 10-30 g/L sucrose, 20-40 g/L sodium chloride, 4-10 g/L-glutamic acid and 0.2-0.8 g/L pyridoxal phosphate.
8. The method according to claim 6, wherein the fermentation conditions are: the fermentation temperature is 25-32 ℃, the pH value in the fermentation process is controlled to be 4.5-6.5, the rotating speed of a fermentation tank is 200-800 rpm, the amount of dissolved oxygen in the fermentation liquid is 20-80%, and the fermentation time is 48-72 h.
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| CN116144516A (en) * | 2023-03-10 | 2023-05-23 | 江南大学 | Saccharomyces cerevisiae for producing succinic acid and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030228567A1 (en) * | 2001-10-26 | 2003-12-11 | Imandokht Famili | Compostions and methods for modeling Saccharomyces cerevisiae metabolism |
| JP2005015348A (en) * | 2003-06-24 | 2005-01-20 | Asahi Denka Kogyo Kk | Skin cosmetic |
| CN104263665A (en) * | 2014-09-23 | 2015-01-07 | 南京工业大学 | Lignin-tolerant saccharomyces cerevisiae and application of lignin-tolerant saccharomyces cerevisiae in production of bioethanol |
| CN110373338A (en) * | 2019-08-15 | 2019-10-25 | 东莞东阳光药物研发有限公司 | Saccharomyces cerevisiae and application thereof |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030228567A1 (en) * | 2001-10-26 | 2003-12-11 | Imandokht Famili | Compostions and methods for modeling Saccharomyces cerevisiae metabolism |
| JP2005015348A (en) * | 2003-06-24 | 2005-01-20 | Asahi Denka Kogyo Kk | Skin cosmetic |
| CN104263665A (en) * | 2014-09-23 | 2015-01-07 | 南京工业大学 | Lignin-tolerant saccharomyces cerevisiae and application of lignin-tolerant saccharomyces cerevisiae in production of bioethanol |
| CN110373338A (en) * | 2019-08-15 | 2019-10-25 | 东莞东阳光药物研发有限公司 | Saccharomyces cerevisiae and application thereof |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116144516A (en) * | 2023-03-10 | 2023-05-23 | 江南大学 | Saccharomyces cerevisiae for producing succinic acid and application thereof |
| CN116144516B (en) * | 2023-03-10 | 2023-08-08 | 江南大学 | Saccharomyces cerevisiae for producing succinic acid and application thereof |
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