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Engineering colon bacillus for preparing biological gasoline

A technology of Escherichia coli and biogasoline, applied in the direction of microorganism-based methods, microorganisms, petroleum industry, etc., can solve the problems of poor quality, easy solidification, and high viscosity of biodiesel, and achieve the effect of easy separation

Active Publication Date: 2010-12-01
青岛生物能源与过程研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the relatively mature bio-liquid fuels include bio-ethanol and bio-diesel (fatty acid methyl ester), but ethanol has low energy density, is easy to absorb water, and is inconvenient to store and transport, while bio-diesel has a high viscosity and is easy to solidify, which will cause diesel engine Filters are prone to clogging and poor start-up performance at low temperatures (Atsumi et al., 2008; Li Chunmei et al., 2008)

Method used

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  • Engineering colon bacillus for preparing biological gasoline
  • Engineering colon bacillus for preparing biological gasoline

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030]By co-overexpressing the acetyl-CoA carboxylase gene (accABCD) of A. calcoaceticus, the BTE gene of U. californica, the acr1 gene of A. calcoaceticus and the CER1 gene of A. thaliana in E. coli, and knocking out the E. coli The fadE gene is used in the biosynthesis of short- and medium-chain alkanes.

[0031] 1.1) Cloning of foreign genes and construction of expression vectors

[0032] 1.1.1) Cloning of foreign genes

[0033] 1.1.1.1) Cloning of A. calcoaceticus acetyl-CoA carboxylase gene

[0034] A. calcoaceticus genomic DNA was extracted, primers were designed according to the GenBank sequence, and the following genes were amplified by PCR:

[0035] accA: acetyl-coenzyme A carboxylase carboxyltransferase (alpha subunit) GeneID (NCBI): 2878570;

[0036] accBC (including accB and accC genes): accB: biotin carboxyl carrier protein of acetyl-CoA, GeneID (NCBI): 2878571; accC: acetyl-CoA carboxylase, GeneID (NCBI): 2878572;

[0037] accD: acetyl-CoA carboxylase, beta s...

Embodiment 2

[0079] Cc FatB of C. calophylla by co-overexpressing the acetyl-CoA carboxylase gene (accABCD) of A. calcoaceticus in E. coli 2 genes, the acr1 gene of A. calcoaceticus and the CER1 gene of A. thaliana, and the fadE gene of E. coli was knocked out for the biosynthesis of short and medium chain alkanes. Method process is the same as embodiment 1.

Embodiment 3

[0081] Ch FatB of C. hookeriana by co-overexpressing the acetyl-CoA carboxylase gene (accABCD) of A. calcoaceticus in E. coli 2 genes, the acr1 gene of A. calcoaceticus and the CER1 gene of A. thaliana, and the fadE gene of E. coli was knocked out for the biosynthesis of short and medium chain alkanes. Method process is the same as embodiment 1.

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Abstract

The invention discloses engineering colon bacillus for preparing intermediate carbon fatty alcohol and a method for preparing the intermediate carbon fatty alcohol. According to the method, the colon bacillus obtains a capability of producing biological gasoline by excessively expressing sulfatase genes, fatty acyl-CoA reductase genes and fatty aldehyde decarboxylase and the like in the colon bacillus, and the biological gasoline producing capability of the engineering colon bacillus is further improved by combining the excessively expressed acetyl-CoA carboxylase genes and knocking out the fatty acyl-CoA dehydrogenase genes of the colon bacillus. The method can be used for producing the new generation biological gasoline.

Description

technical field [0001] The invention relates to an engineering escherichia coli for preparing biogasoline. [0002] The present invention also relates to a method for preparing biogasoline by utilizing the above-mentioned engineering E. coli. Background technique [0003] With the dwindling of oil resources and the aggravation of environmental pollution, bio-liquid fuel, as a clean and renewable energy, is attracting the attention and attention of various countries in the world. At present, the relatively mature bio-liquid fuels include bio-ethanol and bio-diesel (fatty acid methyl ester), but ethanol has low energy density, is easy to absorb water, and is inconvenient to store and transport, while bio-diesel has a high viscosity and is easy to solidify, which will cause diesel engine Filters are prone to clogging and poor start-up performance at low temperatures (Atsumi et al., 2008; Li Chunmei et al., 2008). Therefore, it is necessary to develop a new generation of bio-l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/70C12P5/00C10L1/04C12R1/19
Inventor 咸漠杨建明郑艳宁刘炜徐鑫
Owner 青岛生物能源与过程研究所
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