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Method for biologically synthesizing fatty alcohol from fatty acyl ACP (acyl carrier protein) reductase

A biosynthesis and reductase technology, applied in biochemical equipment and methods, oxidoreductase, botanical equipment and methods, etc., can solve problems such as consumption of oil reserves, achieve excellent anti-pollution performance, facilitate genetic manipulation, reduce The effect of consumption

Active Publication Date: 2012-12-19
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, fatty alcohols that rely on petroleum products as raw materials also consume a large amount of petroleum reserves. Today, when resources are increasingly scarce, this development method will gradually be eliminated. Therefore, a more environmentally friendly and renewable production method is needed

Method used

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  • Method for biologically synthesizing fatty alcohol from fatty acyl ACP (acyl carrier protein) reductase
  • Method for biologically synthesizing fatty alcohol from fatty acyl ACP (acyl carrier protein) reductase
  • Method for biologically synthesizing fatty alcohol from fatty acyl ACP (acyl carrier protein) reductase

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Effect test

Embodiment 1

[0039] The published DPW gene sequence (SEQ ID No.1, synthesized by GenScript Biotechnology Co., Ltd.), and cloned into pET28 vector through the two restriction sites of EcoR Ⅰ and Xho Ⅰ, constructed a good plasmid It was named pRL108.

[0040] Knockout of the fatty acyl carrier protein dehydrogenase fadE gene in BL21 (DE3) can reduce the consumption of fatty acyl-CoA, thereby accumulating more fatty acyl carrier proteins and fatty acyl carrier proteins for the production of fatty alcohols. In this example, Escherichia coli BL21 (DE3) with the fadE gene knocked out was named TL101. Knock out the fatty acyl-CoA synthase fadD gene in TL101, which can interrupt the formation of fatty acyl-CoA, so that only fatty acyl carrier protein is synthesized in the cell. In this example, according to the principle of homologous recombination, a knockout plasmid was constructed to knock out the fadD gene in TL101. Through two primers pRL1-S(TTAAGCATGC GAAGATTTTA CTGCGGATAT T(SphⅠ)) and pRL1-A...

Embodiment 2

[0057] The published AAR gene sequence was optimized and fully synthesized (SEQ ID No.3, synthesized by Jinweizhi Biotechnology Co., Ltd.), and cloned into pET28 vector through Nco Ⅰ and BamH Ⅰ restriction sites, The constructed plasmid was named pFASR.

[0058] The pFASR was transformed into BL21(DE3), and the successful transformant was selected by kanamycin, and named RL12.

[0059] The pFASR was transformed into MG1655(DE3)ΔRECAΔENDA, and the successful transformant was selected by kanamycin and named RL13.

[0060] The pFASR will be transformed into TL101, and the successful transformant will be screened by kanamycin and named RL14.

[0061] The pFASR was transformed into RL101, and the successful transformant was screened by kanamycin and named RL15.

[0062] The pFASR and pMSD8 were co-transformed into TL101, and the successful transformant was selected by Karamycin and named RL16.

[0063] The pFASR and pMSD8 were co-transformed into RL101, and the successful transformant was sc...

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Abstract

The invention discloses a method for biologically synthesizing fatty alcohol from fatty acyl ACP (acyl carrier protein) reductase. According to the method, fatty acyl carrier protein reductase genes are guided into a host cell body, and the genes are expressed in the host cell body, so the fatty acid metabolism pathway in heterotrophic microorganisms or heterotrophic cell bodies is transformed for producing the fatty alcohol. The method also further improves the fatty alcohol yield through metabolism transformation. The method has the advantages that the fatty alcohol synthesis in micro-organisms by a biosynthesis method is realized, the technology is a renewable low-consumption technology, in addition, the consumption of scarce resources of petroleum can be reduced in the environment-friendly production mode, the chemical modification is also not needed, the fatty alcohol can be directly produced, and great industrial application prospects are realized.

Description

Technical field [0001] The invention belongs to the field of renewable energy, biomass energy and chemical raw material production, and specifically relates to a method for modifying the fatty acid metabolism pathway of heterotrophic microorganisms to produce fatty alcohols. Background technique [0002] Fatty alcohols are one of the raw materials of surfactants for detergents, and they are widely used in detergents, skin care products, cosmetics, and medicines. Fatty alcohol was first produced from spermaceti. The resulting mixed fatty alcohol was sulfonated and neutralized to become sulfate, which was the earliest anionic detergent. Later, the rich sources of coconut oil, palm oil and tallow were developed and used as raw materials, and the fatty acids obtained by hydrolysis were reduced to alcohols, collectively referred to as natural fatty alcohols. After the development of the petrochemical industry, petroleum products are used as raw materials to produce fatty alcohols cal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/02C12N1/15C12N1/19C12N1/21C12N5/10C12N15/53C12N9/02
Inventor 刘天罡刘然付爱思
Owner WUHAN UNIV
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