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Method for fixing carbon dioxide in situ in ethanol fermentation process

An ethanol fermentation and carbon dioxide technology, applied in the biological field, can solve the problem that carbon dioxide fixation efficiency needs to be further improved, and achieve the effects of improving carbon source utilization rate, speeding up fermentation rate, and improving ethanol yield

Pending Publication Date: 2022-05-27
DALIAN UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Formate dehydrogenase (FDH) can reduce carbon dioxide to soluble formic acid, which has been used in the research of in vitro electrochemical method to fix carbon dioxide, but in vitro carbon dioxide fixation requires electrolysis of water to provide additional energy, and the efficiency of carbon dioxide fixation remains to be seen. to further improve

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  • Method for fixing carbon dioxide in situ in ethanol fermentation process
  • Method for fixing carbon dioxide in situ in ethanol fermentation process
  • Method for fixing carbon dioxide in situ in ethanol fermentation process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Construction of strain S. cerevisiae-FDH

[0025] The Fdh gene fragment was obtained by PCR amplification with the genome of Kluyveromyces marxianus as the template. The PCR amplification kit was HSDNA Polymerase (Code No.: R010A) purchased from Bao Bioengineering Dalian Co., Ltd. The PCR reaction was carried out according to the kit instructions . The PCR amplification conditions were denaturation at 99°C for 5 min, 98°C for 15s, 60°C for 15s, 72°C for 60s for 30 cycles, and extension at 72°C for 7 min. The obtained Fdh gene fragment was double-digested with restriction endonucleases Not I and Sal I, and then received On pRS424, the expression vector pRS424-FDH was obtained, and then transformed into Saccharomyces cerevisiae S288C to obtain the recombinant strain S.cerevisiae-FDH overexpressing formate dehydrogenase

Embodiment 2

[0026] Example 2: Fermentation effect and formic acid yield under 20g / L glucose

[0027] 1) Preparation of activation medium: activation medium: YNB 6.7 g / L, glucose 10 g / L, amino acid supplement solution without Trp.

[0028]2) Activation of strains: S. cerevisiae-FDH and the control strain S. cerevisiae S288C stored in the refrigerator were inoculated into the activation medium, placed in a shaker at 30° C. and cultured at 150 rpm for 48 hours.

[0029] 3) Preparation of seed medium: Seed medium: YNB 6.7 g / L, glucose 20 g / L, amino acid supplemented solution without Trp.

[0030] 4) In step 2), the activated bacterial cells were transferred to the seed medium prepared in step 3), placed in a shaker at 30° C. and cultured at 150 rpm for 48 hours.

[0031] 5) Preparation of fermentation medium: YNB 6.7 g / L, glucose 20 g / L, amino acid supplement solution without Trp.

[0032] 6) After the strains are activated and the seeds are expanded and cultured, the cells are collected by...

Embodiment 3

[0038] Example 3: Inoculum OD 620 Fermentation effect and formic acid yield under =1

[0039] 1) Preparation of activation medium: activation medium: YNB 6.7 g / L, glucose 10 g / L, amino acid supplement solution without Trp.

[0040] 2) Activation of strains: S. cerevisiae-FDH and the control strain S. cerevisiae S288C stored in the refrigerator were inoculated into the activation medium, placed in a shaker at 30° C. and cultured at 150 rpm for 48 hours.

[0041] 3) Preparation of seed medium: Seed medium: YNB 6.7 g / L, glucose 20 g / L, amino acid supplemented solution without Trp.

[0042] 4) In step 2), the activated bacterial cells were transferred to the seed medium prepared in step 3), placed in a shaker at 30° C. and cultured at 150 rpm for 48 hours.

[0043] 5) Preparation of fermentation medium: identification medium: YNB 6.7 g / L, glucose 40 g / L, amino acid supplement solution without Trp.

[0044] 6) After the strains are activated and the seeds are expanded and cultur...

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Abstract

The invention belongs to the technical field of biology, and discloses a method for in-situ immobilization of carbon dioxide in an ethanol fermentation process, which comprises the following steps: (1) constructing recombinant Saccharomyces cerevisiae-FDH for expressing formate dehydrogenase; (2) determining the carbon dioxide fixed amount and the ethanol yield of the recombinant strain S.cerevisiae-FDH under different glucose concentrations; (3) the carbon dioxide fixed amount and the ethanol yield of the recombinant strain S.cerevisiae-FDH under different inoculum sizes are determined; and (4) adding a carbon dioxide cosolvent such as 2-methylimidazole zinc salt MAF-4 to improve the fixing efficiency of carbon dioxide. The method is simple and easy to implement, does not affect the main fermentation process and microbial growth, utilizes carbon dioxide released in the ethanol fermentation process in situ, and achieves the purposes of carbon dioxide emission reduction and total carbon recovery rate improvement.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for in-situ fixation of carbon dioxide in ethanol fermentation, which utilizes formate dehydrogenase in microbial cells to in-situ convert carbon dioxide produced in the process of ethanol fermentation and can speed up the fermentation rate and improve the target product yield. Background technique [0002] Since the Industrial Revolution, the rapid development of heating, power generation, transportation and other industries has emitted a large amount of greenhouse gases, resulting in serious climate change and global warming, which seriously threatens human society. For the sustainable development of human society, it is necessary to reduce the concentration of carbon dioxide in the air. Countries around the world have not only implemented global regulations such as the Paris Climate Agreement to reduce carbon dioxide emissions, but are also developing various capture and ut...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/81C12N15/53C12N1/19C12P7/06C12R1/865
CPCC12N15/81C12N9/0008C12P7/06C12Y102/01002Y02E50/10
Inventor 袁文杰杜聪李益民
Owner DALIAN UNIV OF TECH
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