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Method for improving carbon monoxide utilization rate of clostridium ljungdahlii by high-throughput gene synthesis process

A carbon monoxide, utilization technology, applied in microorganism-based methods, chemical instruments and methods, biochemical equipment and methods, etc., can solve problems such as low solubility

Active Publication Date: 2021-02-02
南京食气生化科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007]However, the solubility of CO in water is low (only 77% of oxygen), so the gas-liquid mass transfer rate of CO is the cell density that determines the microbial fermentation of syngas and the important limiting factor of product yield

Method used

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  • Method for improving carbon monoxide utilization rate of clostridium ljungdahlii by high-throughput gene synthesis process
  • Method for improving carbon monoxide utilization rate of clostridium ljungdahlii by high-throughput gene synthesis process
  • Method for improving carbon monoxide utilization rate of clostridium ljungdahlii by high-throughput gene synthesis process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1: Screening of different sources of hemoglobin in Clostridium ljungdahlii

[0067] experimental method:

[0068] Nine hemoglobin genes from different sources were overexpressed using Clostridium universal artificial promoters, and the effects of hemoglobin on the fermentation phenotype of the strains were identified.

[0069] Experimental steps:

[0070] 1) Select the hemoglobin that has been used for engineering in microorganisms, or the hemoglobin that has been reported to have a strong binding force to carbon monoxide, and select 9 hemoglobins as target proteins for preliminary screening, as shown in Table 1.

[0071] 2) The synthetic genes of the above nine kinds of hemoglobin were constructed on the pMTL83151 vector, and the genes were expressed with a strong artificial promoter Pthl9.

[0072] 3) Transforming the above-mentioned vector containing the hemoglobin expression gene into Clostridium ljungdahlii. The obtained engineered bacteria feed carbon m...

Embodiment 2

[0078] Example 2: Screening out the hemoglobin that contributes the most to the fermentation phenotype

[0079] experimental method:

[0080] The two recombinant strains with the best performance in step 1 were selected for further verification of gas fermentation, and the utilization of carbon monoxide in the gas was measured.

[0081] Experimental steps:

[0082] 1) Sequence the total amount of acetate and ethanol produced at the fermentation end point (96h) of different recombinant strains, and select the two strains with the highest total amount of solvents that overexpress CGb and VHb for further confirmation.

[0083] 2), activate the selected recombinant strain in 1) on the YTF plate, carry out gas fermentation, and monitor the growth phenotype.

[0084] 3), in the fermentation course of 2), take the fermentation sample liquid of different time points, measure the output of acetic acid and ethanol by high-efficiency gas chromatography. The residual gas samples at dif...

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Abstract

The invention provides a method for improving the carbon monoxide utilization rate of clostridium ljungdahlii by a high-throughput gene synthesis process. Specifically, the invention provides engineered clostridium ljungdahlii. The carbon monoxide utilization rate of the engineered clostridium ljungdahlii is greater than or equal to 30.0%, preferably greater than or equal to 37.5%, and more preferably greater than or equal to 40.0%. Moreover, the invention provides a preparation method and application of the engineered clostridium ljungdahlii, and a biological carbon fixation method for efficiently utilizing carbon monoxide. According to the technical scheme provided by the invention, the growth, fermentation phenotype and carbon utilization rate of the clostridium ljungdahlii are successfully improved, and a new thought is provided for subsequent metabolic engineering design and transformation of the clostridium ljungdahlii.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a method for improving the carbon monoxide utilization rate of Clostridium aerogenes by a high-throughput gene synthesis method. Background technique [0002] In the microbial fermentation industry, sugar-based raw materials have always been an important source of substrates. With the demand for sustainable development, newer and cheaper biomass raw materials have broader prospects as industrial fermentation raw materials for recombinant microorganisms. [0003] In recent years, carbon-gas has attracted extensive attention. Carbon-gas is an important carbon resource with a wide range of sources and low price, including CO, CO 2 and CH 4 etc., mainly come from waste gas from large-scale petrochemical and smelting enterprises, and synthesis gas produced by gasification of carbonaceous substances such as coal, oil, natural gas, and biomass. Therefore, the resource...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/74C12P7/54C12P7/08C12R1/145
CPCC07K14/805C12N15/74C12P7/54C12P7/08Y02E50/10
Inventor 顾阳杨高华姜卫红
Owner 南京食气生化科技有限公司