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Stress-resistant protective agent based on alginate oligosaccharides as well as preparation method and application of stress-resistant protective agent

A technology of sodium alginate oligosaccharides and stress-resistant protective agents, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, and preservation of microorganisms. It can solve the problems of Saccharomyces cerevisiae fermentation and environmental protection under adversity, and shorten the fermentation period. time, increase fermentation intensity, and increase cell viability

Active Publication Date: 2016-05-04
DALIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In recent years, the biological activity of sodium alginate oligosaccharides has been widely concerned by scholars at home and abroad. Studies have shown that sodium alginate oligosaccharides have a variety of biological activities, such as promoting plant growth and inducing stress resistance, antioxidant activity, and antibacterial activity. activity, inducing the production of animal cytokines, etc., but its application to Saccharomyces cerevisiae fermentation and adversity environmental protection has not been reported

Method used

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  • Stress-resistant protective agent based on alginate oligosaccharides as well as preparation method and application of stress-resistant protective agent
  • Stress-resistant protective agent based on alginate oligosaccharides as well as preparation method and application of stress-resistant protective agent
  • Stress-resistant protective agent based on alginate oligosaccharides as well as preparation method and application of stress-resistant protective agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Preparation of stress-resistant protective agent

[0031] (1) Sodium alginate is dissolved in buffer as a reaction substrate, the concentration of sodium alginate is 1.5% (w / v), and hydrochloric acid with a concentration of 0.3% (v / v) is added thereto, and the degradation temperature is 90°C , the degradation time is 24 hours;

[0032] (2) After the degradation is complete, centrifuge to retain the supernatant and neutralize it to neutrality. Use a dialysis bag with a molecular weight cut-off of 100D to remove the salt in the supernatant, and then freeze-dry the desalinated supernatant to obtain sodium alginate Oligosaccharide powder;

[0033] The reducing sugar content and the total sugar content in the obtained sodium alginate oligosaccharide powder were measured by conventional methods, and the average relative molecular mass was calculated to be about 2497, and the average degree of polymerization of the sodium alginate oligosaccharide was about 6.

[0034] (3) So...

Embodiment 2

[0036] Application of Stress Resistant Protective Agent in Saccharomyces cerevisiae Fermentation

[0037] Saccharomyces cerevisiae Sc4126 in the logarithmic growth phase was inoculated at 1% (v / v) in high-sugar fermentation medium (formulation of high-sugar fermentation medium: 265g / L glucose, 8g / L peptone, 6g / L yeast powder) , and then add anti-stress protective agent with a concentration of 10ug / mL and 10mg / mL to this medium, stir evenly, ferment at 30°C and 150rpm for 48h, measure residual sugar and ethanol every 6h; group is the negative control group.

[0038] Test results such as Figure 1-2 As shown, the end time of fermentation in the group added with the anti-stress protection agent was shortened by 2-6 hours compared with the negative control group, and the ethanol yield at the end point was the same in each group, all around 0.46. The formula for calculating ethanol yield is as follows:

[0039] Ethanol yield = endpoint ethanol mass / initial sugar concentration. ...

Embodiment 3

[0041] Application of stress-resistance protective agent under the stress condition of Saccharomyces cerevisiae

[0042] Take the overnight culture of Saccharomyces cerevisiae Sc4126 in the late logarithmic growth phase to the early stationary phase, centrifuge at 3000r / min for 5min, discard the supernatant, and wash the precipitate with 0.1mol / L trisodium citrate buffer solution of pH5.0 Twice, finally add trisodium citrate buffer to redissolve the precipitate, measure its OD600 value at any time until its OD600 value reaches about 2.0, then add the concentration prepared in Example 1 at 1ug / mL, 10ug / mL or 10mg / mL Stress-resistant protective agent, cultured at 30°C for 2 hours, then centrifuged to remove the stress-resistant protective agent, resuspended Saccharomyces cerevisiae Sc4126 cells with trisodium citrate buffer; exposed Saccharomyces cerevisiae Sc4126 cells to different stress-resistant conditions , stress resistance conditions include:

[0043] Treat the cells wit...

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Abstract

The invention relates to a stress-resistant protective agent for saccharomyces cerevisiae, in particular to a stress-resistant protective agent based on alginate oligosaccharides as well as a preparation method and an application of the stress-resistant protective agent. The stress-resistant protective agent mainly comprises the alginate oligosaccharides and a solvent, wherein the mass concentration of the alginate oligosaccharides is 1 mu g / mL-10mg / mL; the alginate oligosaccharides comprise mannuronate, guluronic acid or a hybrid of mannuronate and guluronic acid, the average degree of polymerization is 5-8, and the average molecular weight ranges from 2,000 to 3,000. The stress-resistant protective agent is added to a culture medium of saccharomyces cerevisiae, the application field of marine-derived polysaccharides and oligosaccharides is expanded, the high-glucose fermentation time is shortened, the fermentation strength is improved, and the survival rate and the cell activity of saccharomyces cerevisiae in multiple stress environments are increased.

Description

technical field [0001] The invention relates to the field of stress tolerance of industrial microorganisms and the field of marine polysaccharide / oligosaccharide bioutilization, in particular to a stress-resistant protective agent for Saccharomyces cerevisiae. Background technique [0002] Saccharomyces cerevisiae has been widely used in the fields of bioenergy and food and medicine (wine making, pasta food, biomedicine, etc.) because of its excellent wine production performance and good biological safety. Especially in recent years, the production and use of fuel ethanol as a renewable clean energy has attracted widespread attention. Among them, the production of fuel ethanol using non-grain cellulose instead of grain can effectively solve or alleviate the food crisis and alleviate the contradiction of oil resource shortage. , At the same time, it is also conducive to the improvement of the environment and the reduction of air pollution. [0003] However, in the fermentati...

Claims

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Application Information

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IPC IPC(8): C12N1/18C12N1/04C12P7/06C08B37/04C12R1/865
CPCC08B37/0084C12N1/04C12N1/18C12P7/06Y02E50/10
Inventor 吴海歌李倩姚子昂崔红利秦丽媛高晨慧吴晨
Owner DALIAN UNIV
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