Method for biosynthesizing fatty alcohol by using fatty acyl ACP (acyl carrier protein) reductase

A technology of biosynthesis and reductase, which is applied in the field of modifying the fatty acid metabolism pathway of heterotrophic microorganisms to produce fatty alcohols. It can solve the problems of oil reserve consumption, achieve excellent anti-pollution performance, facilitate genetic manipulation, and reduce pollution.

Pending Publication Date: 2012-10-10
WUHAN UNIV
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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 incr

Method used

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  • Method for biosynthesizing fatty alcohol by using fatty acyl ACP (acyl carrier protein) reductase
  • Method for biosynthesizing fatty alcohol by using fatty acyl ACP (acyl carrier protein) reductase
  • Method for biosynthesizing fatty alcohol by using fatty acyl ACP (acyl carrier protein) reductase

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Embodiment 1

[0037] will be announced DPW Gene sequence (SEQ ID No.1, synthesized by GenScript Biotechnology Co., Ltd.), and passed Eco RI and xho Ⅰ Cloning it into the pET28 vector with two restriction sites, and the constructed plasmid was named pRL108.

[0038] The acyl carrier protein dehydrogenase in BL21(DE3) fad E Gene knockout 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 instance will knock out fad E The gene of Escherichia coli BL21(DE3) was named TL101. Fatty acyl-CoA synthetase in TL101 fadD Gene knockout 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. Through two primers pRL1-S(TTAA GCATGC GAAGATTTTA CTGCGGATAT ( Sph Ⅰ)) and pR...

Embodiment 2

[0055] will be announced AAR The gene sequence was optimized and fully synthesized (SEQ ID No.3, synthesized by Jinweizhi Biotechnology Co., Ltd.), and passed Nco I and Bam HI two restriction sites to clone it into pET28 vector, and the constructed plasmid was named pFASR.

[0056] The pFASR was transformed into BL21(DE3), and a successful transformant was obtained by screening with kanamycin, which was named RL12.

[0057] Transformation of pFASR into MG1655(DE3) ΔRECAΔENDA , using kanamycin to select a successful transformant, which was named RL13.

[0058] The pFASR will be transformed into TL101, and the successful transformants obtained by kanamycin screening will be named RL14.

[0059] The pFASR was transformed into RL101, and a successful transformant was obtained by kanamycin screening, which was named RL15.

[0060] The pFASR and pMSD8 were co-transformed into TL101, and the successful transformant was screened by karimycin, which was named RL16.

[0061] T...

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Abstract

The invention discloses a method for biosynthesizing fatty alcohol by using a fatty acyl ACP (acyl carrier protein) reductase. The method comprises the following steps of: introducing a fatty acyl carrier protein reductase gene into the body of a host cell, expressing the gene in the body of the host cell, and thus transforming a metabolizing way of fatty acid in the body of a heterotrophic microorganism or a heterotrophic cell so as to produce fatty alcohol. By the method, the yield of the fatty alcohol is further improved through metabolic transformation and the fatty alcohol is synthesized in the body of the microorganism through biosynthesis; the method is a renewable and low-consumption technology; by such environment-friendly method, the consumption of scarce petroleum resource can be reduced and the fatty alcohol can be directly produced without chemical transformation; and the method has a wide industrial application prospect.

Description

technical field [0001] The invention belongs to the fields of renewable energy, biomass energy and chemical raw material production, and specifically relates to a method for producing fatty alcohol by transforming the fatty acid metabolism pathway of heterotrophic microorganisms. Background technique [0002] Fatty alcohols are used in detergents Surfactant It is widely used in detergents, skin care products, cosmetics and medicines. Fatty alcohols were first produced from cetyl wax, and the resulting mixed fatty alcohols became sulfates after sulfonation and neutralization, which is the earliest Anionic detergent . Later, it developed and utilized coconut oil, palm oil and tallow with abundant sources as raw materials. hydrolysis The resulting fatty acids are then reduced to alcohols, collectively referred to as natural fatty alcohols. After the development of the petrochemical industry, the fatty alcohols produced from petroleum products as raw materials are called s...

Claims

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Application Information

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IPC IPC(8): C12N15/63C12N1/21C12N1/13C12N1/15C12N1/19C12N5/10C12P7/04C12R1/19
Inventor 刘天罡刘然付爱思
Owner WUHAN UNIV
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