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Ethanol-tolerant, high-temperature-tolerant and high-permeability-tolerant multi-tolerance fermentation strain and construction method

A technology for fermenting strains and construction methods, applied in the direction of microorganism-based methods, biochemical equipment and methods, fungi, etc., which can solve the problems of easily changing performance, not suitable for industrial production conditions, unsatisfactory fermentation ability of multi-tolerant strains, etc. question

Active Publication Date: 2021-01-08
CHINA PETROLEUM & CHEM CORP +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, most of the researches use laboratory yeast whose properties are easy to change as the original strain, which is not suitable for industrial p

Method used

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  • Ethanol-tolerant, high-temperature-tolerant and high-permeability-tolerant multi-tolerance fermentation strain and construction method
  • Ethanol-tolerant, high-temperature-tolerant and high-permeability-tolerant multi-tolerance fermentation strain and construction method
  • Ethanol-tolerant, high-temperature-tolerant and high-permeability-tolerant multi-tolerance fermentation strain and construction method

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Experimental program
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Effect test

Embodiment 1

[0037] refer to figure 1 , the present embodiment provides a method for constructing a multi-tolerant fermentation strain resistant to ethanol, high temperature and hypertonicity, comprising the steps of:

[0038] Step 1, the construction of the starting strain

[0039]The spores produced by SEB1 and IR-2 were fused by protoplasts to obtain flocculant fusogen RHZ-1, of which SEB1 was preserved in the General Microbiology Center of China Microbiological Culture Collection Management Committee, and the preservation number was CGMCC NO.11321; the IR-2 strain was isolated Fermented food from Indonesia, with ethanol production capacity and flocculation, the source is described in the following documents: Hiroshi K, Yoshio S, Toshio M, Harumi K, Yorikazu S.1985.Continuous ethanolfermentation with cell recycling using flocculatingyeast.J.Ferment .Technol.63:159-165; the translated name of this document is: use flocculating yeast to carry out continuous ethanol fermentation with cell...

Embodiment 2

[0053] This embodiment verifies the fermentation performance of the multi-tolerant bacterial strain SEB14 under high temperature conditions, and the specific verification process and results are as follows:

[0054] Under the fermentation conditions of 42°C and 43°C (10% YPD), both strains RHZ-1 and SEB14 could consume glucose within 24h and 48h, indicating that the starting strain RHZ-1 itself has better high temperature tolerance. When the fermentation temperature increased to 44 °C, the ethanol concentration of SEB14 was significantly higher than that of RHZ-1 ( image 3 ). After 72 hours of fermentation, the ethanol concentration and glucose concentration of SEB14 were 40.98g / L and 10.71g / L, respectively. Compared with RHZ-1, it increased by 33.36% and decreased by 66.72% (Table 2). This result shows that SEB14 has stronger high temperature resistance.

[0055] Table 2 Comparison of strains fermented for 72 hours at 44°C

[0056]

[0057] Note: The same letter in th...

Embodiment 3

[0059] This example verifies the fermentation performance of the multi-tolerant strain SEB14 at high temperature and contains ethanol. The specific verification process and results are as follows:

[0060] In the late stage of SSF fermentation, the yeast should not only tolerate high temperature, but also gradually increase the concentration of ethanol at the same time. In this study, we found that SEB14 also exhibited better fermentation performance than the parental strain RHZ-1 under the co-fermentation conditions of initial addition of 3% ethanol and high temperature at 43°C, indicating that the strain SEB14 can well withstand the dual effects of high temperature and ethanol. Coercion ( Figure 4 ,table 3).

[0061] Table 3 Comparison of results of 72h fermentation of strains with initial addition of 3% ethanol and 43°C

[0062]

[0063] Note: The same letter in the same column means no significant difference, different letters mean significant difference, P<0.05(t-te...

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Abstract

The invention provides an ethanol-tolerant, high-temperature-tolerant and high-permeability-tolerant multi-tolerance fermentation strain and a construction method. The saccharomyces cerevisiae strainis SEB14, is classified and named as saccharomyces cerevisiae, and has a preservation number of CGMCC No. 19587. An industrial saccharomyces cerevisiae strain RHZ-1 is used as a starting strain, ARTPmutation and multiple rounds of genome rearrangement are carried out on the industrial saccharomyces cerevisiae strain RHZ-1, and mutant strains are screened through TTC chromogenic reaction, Duchennecatheters, triangular flask fermentation and the like, so that the excellent multi-tolerance strain SEB14 is finally obtained. The SEB14 has better high temperature tolerance, ethanol tolerance, highsugar tolerance and high permeability tolerance, and has better industrial application potential.

Description

technical field [0001] The invention relates to the technical field of microbial recombination, in particular to a multi-tolerant fermentation strain with ethanol resistance, high temperature resistance and hypertonic resistance and a construction method. Background technique [0002] The annual production of bioethanol has exceeded 50 billion liters, and it has become an eco-friendly renewable energy. Improving the production and economic benefits of bioethanol can promote its further application and development. In the current industrial fermentation, very high concentration (VHG) ethanol fermentation (the sugar content of the medium is greater than 250g / L in order to obtain more than 15% ethanol) and simultaneous saccharification and fermentation (SSF) are considered to be effective, energy-saving, and Widely used industrial ethanol production process. However, in industrial production, many environmental stresses, including ethanol, high temperature, and osmotic pressur...

Claims

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Application Information

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IPC IPC(8): C12N1/16C12N15/04C12N15/01C12R1/865
CPCC12N1/16C12N15/04C12N15/01C12R2001/865C12N1/185Y02E50/10
Inventor 汤岳琴陈栋王莉缪晡宋清冉
Owner CHINA PETROLEUM & CHEM CORP
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