Method for producing yeast extracellular trehalose by two step fermentation method

A technology of trehalose and fermentation method is applied in the field of producing yeast extracellular trehalose, which can solve the problems of up to 1.5 to 1.8 dollars, and achieve the effects of improving food quality, reducing production cost and high conversion rate.

Inactive Publication Date: 2003-11-26
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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AI Technical Summary

Problems solved by technology

If it is used as a food protective agent, its effective protective effect should be added about 15%, and the cost of the protective agent per kilogram of food will be as high as 1.5 to 1.8 US dollars

Method used

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  • Method for producing yeast extracellular trehalose by two step fermentation method
  • Method for producing yeast extracellular trehalose by two step fermentation method

Examples

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Embodiment 1

[0039] Example 1: Trehalose synthesis test in a 500mL Erlenmeyer flask, as shown in Figure 1:

[0040] 1. Stressed yeast cell culture: In a 250mL Erlenmeyer flask with 50mL of compound medium, inoculate 1-2 rings of the preserved strains on the slant, and culture at a constant temperature of 30°C for 24-36h, as the primary seed. Then put 100mL nitrogen-limited medium into a 500mL Erlenmeyer flask, add the first-grade seed solution with 5% inoculum, and culture at 30°C with shaking at 150r / min for 16-24h. In the later stage of cultivation, raise the temperature to 36-38°C to start the expression of yeast trehalose synthase gene, then add 2.5% NaCl to the medium to stimulate the formation of yeast trehalose synthase and increase the accumulation of trehalose in yeast . After culturing, centrifuge and wash twice with sterile water before use.

[0041] 2. Synthesis of trehalose: Weigh 2g of the yeast sludge in the stress state, add 10mL of 4% toluene, and shake at 36-38°C for 1 ...

Embodiment 2

[0045] Example 2: Synthesis of trehalose in a 5L self-controlled fermenter as shown in Figure 2:

[0046] 1. Stressed yeast cell fed-batch culture: put the yeast slurry into a sterilized 5L self-controlled fermenter according to the inoculation amount of 2%, the culture temperature is 28 ° C ~ 30 ° C, and 10% Na 2 CO 3 Adjust the pH, add molasses and nutrient salts, so that the fermentable sugar in the medium is kept at 0.1-1% throughout the process, and the final liquid volume is about 3L; the pH is controlled at 3.8-4.5 at the beginning of the culture, and gradually increases after 10 hours After cultivating for a period of time, increase the pH of the fermentation broth to 5.0-7.0; add 2.5% NaCl in the later stage of cultivation, and increase the temperature of the fermentation broth to 36-38°C until the end of the cultivation. Yeast sludge was obtained by centrifugation, washed twice with sterile water, and preserved. At this time, yeast cells have accumulated more stres...

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Abstract

Of the two-step fermentation process of producing yeast extracellular trehalose, the first step is the culture of stress yeast cell to induce yeast cell to produce more stress enzyme-trehalose synthetase series via nitrogen hungry, heat shock, osmotic menace and other technological process; and the second step is the synthesis of trehalose as the stress metabolism product to shock treat the stress yeast cell physical or chemically, to eliminate the limitation of yeast cell membrane in the osmolarity to trehalose and to synthesize great amount of extracellular trehalose with the trahalose synthesizing enzyme system in yeast cell. The process has high yield, simple production process, less investment and low production cost. The product trehalose has excellent non-specific protection to biological macro molecules, and may be used widely in molecular biology, biomedicine product, food, cosmetics and other fields.

Description

Technical field: [0001] The invention relates to a method for producing trehalose, in particular to a method for producing yeast extracellular trehalose by using glucose as a substrate and adopting a two-step fermentation method. Background technique: [0002] It is well known that yeast produces stress metabolite trehalose under extreme conditions such as desiccation, high temperature shock, low temperature freezing and hyperosmotic stress, and its presence can keep yeast cells in a dehydrated state for several years without inactivation. The significance of trehalose research is that it has a good non-specific protective effect on biological macromolecules, making it widely used in molecular biology, pharmaceutical biological products, food and cosmetics and other fields. In molecular biology, with the development of biotechnology, various tool enzymes, probes, antibodies, organelles and biological reagents are more and more widely used. The application of trehalose can ke...

Claims

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Application Information

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IPC IPC(8): C12P19/12
Inventor 肖冬光赵华王兰李于郭波
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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