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Multi-tolerance saccharomyces cerevisiae strain and construction method and application thereof

A Saccharomyces cerevisiae strain and tolerance technology, applied in the field of bioengineering, can solve the problems of stress resistance and single tolerance, achieve the effects of improved tolerance, good industrial application potential, and improved ethanol resistance

Active Publication Date: 2021-11-30
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on the stress resistance of Saccharomyces cerevisiae is mainly focused on the improvement of single tolerance. The main methods of breeding are mutagenesis, domestication, genome rearrangement and genetic engineering, etc. How to improve the multiple tolerance of Saccharomyces cerevisiae at the same time very limited research

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  • Multi-tolerance saccharomyces cerevisiae strain and construction method and application thereof
  • Multi-tolerance saccharomyces cerevisiae strain and construction method and application thereof
  • Multi-tolerance saccharomyces cerevisiae strain and construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0114] Ethanol glucose co-fermentation:

[0115] Using the industrial Saccharomyces cerevisiae strain SEB14 as the host and highly expressing the gene ENA5, an excellent multi-tolerant strain SEB17 was obtained. After co-fermentation with 9% (v / v) initial ethanol and 141.95g / L glucose for 96h, it produced 32.39±1.02g / L ethanol, which was 13.09% (28.64±1.66g / L) higher than that of strain SEB14. Meanwhile, after 24 hours of fermentation, the fermentation rate of SEB17 was significantly higher than that of SEB14. The results indicated that the highly expressed gene ENA5 could significantly improve the ethanol tolerance of SEB14, as shown in Table 11 and figure 1 shown.

[0116] Table 11 96h results comparison of bacterial strain fermentation under the mixed condition of 9% (v / v) ethanol and 141g / L glucose

[0117]

[0118] Note: The same letter in the same column means no significant difference, different letters mean significant difference, P<0.05(t-test). 9% (v / v) ethano...

Embodiment 2

[0120] High temperature fermentation:

[0121] This example explores the high temperature tolerance of strain SEB17. Under the fermentation conditions of 44°C and 100.65g / L glucose, the ethanol concentration of strain SEB17 was significantly (P figure 2 ). After 72 hours of fermentation, the ethanol concentration and glucose concentration of SEB17 were 43.52±1.17g / L and 7.21±2.03g / L, respectively. Compared with SEB14, it increased by 14.17% and decreased by 59.74% (Table 12). This result indicated that high expression of ENA5 could significantly improve the high temperature tolerance of strain SEB14.

[0122] Table 12 Comparison of strains fermented for 72 hours at 44°C

[0123]

[0124] Note: The same letter in the same column means no significant difference, different letters mean significant difference, P<0.05(t-test). SEB17:

[0125] The strain SEB14 highly expressed the gene ENA5.

Embodiment 3

[0127] High temperature ethanol co-fermentation:

[0128] In the late stage of SSF fermentation, yeast cells must tolerate both high temperature and gradually increasing ethanol concentration. In this example, it was found that SEB17 also exhibited better fermentation performance than the starting strain SEB14 under the co-fermentation conditions of initial addition of 3% ethanol and high temperature of 43°C. At the end of the fermentation, its ethanol production concentration (23.43 ± 0.95g / L) increased by 20.36% compared with the starting strain SEB14 (18.66 ± 1.62g / L), indicating that the strain SEB17 can well tolerate the dual stress of high temperature and ethanol, The highly expressed gene ENA5 is crucial for improving the tolerance of strain SEB14 to the dual stress of high temperature and ethanol ( image 3 , Table 13).

[0129] Table 13 Comparison of results of 72h fermentation of strains with initial addition of 3% ethanol and 43°C

[0130]

[0131] Note: The s...

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Abstract

The invention discloses a multi-tolerance saccharomyces cerevisiae strain and a construction method and application thereof. The saccharomyces cerevisiae strain is preserved in the China general microbiological culture Collection center, the preservation name is SEB17, and the preservation number is CGMCC NO. 22587. The construction method comprises the steps that a strain SEB14 is adopted as an original strain, a promoter of a functional gene ENA5 of the strain SEB14 is replaced with a promoter of TEF1, and the strain SEB17 is constructed. The strain SEB17 has the multi-tolerance, has the advantage of being resistant to various environmental stresses such as high ethanol, high temperature, high sugar and high salt, and can be suitable for material fermentation under various environmental stresses.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a multi-tolerant Saccharomyces cerevisiae strain and its construction method and application. Background technique [0002] Facing the increasingly scarce energy and the increasingly serious environmental pollution, it is urgent to develop green and renewable new energy sources. The resource utilization of organic wastes such as straw and waste molasses with large reserves and production volume to produce clean energy fuel ethanol will alleviate the energy and environmental crises at the same time. If ultra-high concentration (VHG) fermentation and high-temperature fermentation can be realized in the industrial production of fuel ethanol, production costs can be significantly reduced, waste water discharge can be reduced, and environmental pollution load can be reduced. Therefore, breeding multi-tolerant industrial Saccharomyces cerevisiae strains that can tolerate multip...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/113C12N15/55C12P7/10C12P7/08C12R1/865
CPCC12N9/14C12P7/10C12P7/08Y02E50/10
Inventor 汤岳琴王莉谢采芸苟敏孙照勇夏子渊
Owner SICHUAN UNIV
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