Application of fatty acid elongase gene in yeast synthesis of nervonic acid

A technology of neuramic acid and Yarrowia lipolytica, which is applied in the fields of application, genetic engineering, and plant genetic improvement, and can solve the problems of insufficient pathogenic production potential of neuric acid-producing microorganisms and inability to industrialize production.

Pending Publication Date: 2022-05-20
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most of the known neuric acid-producing microorganisms are pathogenic or have

Method used

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  • Application of fatty acid elongase gene in yeast synthesis of nervonic acid
  • Application of fatty acid elongase gene in yeast synthesis of nervonic acid
  • Application of fatty acid elongase gene in yeast synthesis of nervonic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0111] Example 1: Yeast expresses MaoleKCS to synthesize nervonic acid

[0112] 1. Materials and Methods

[0113] 1.1 Strains and culture conditions

[0114] The starting strain of Yarrowia lipolytica used in the examples is Y. lipolytica Po1g (MATa, leu2-270, ura3-302::URA3, xpr2-3), purchased from Yeastern Biotech Co., Ltd. The bacterial strain Po1g-G3-ΔURA was obtained by the following method, specifically: the bacterial strain Po1g-G3 was obtained by overexpressing the endogenous genes DGAT1, ACC1 and SCD genes of Yarrowia lipolytica (Qiao and Stephanopoulos.Metabolic Engineering, 2015, 29 :56-65), and then reverse screening by 5-FOA method to obtain uracil auxotrophic strain Po1g-G3-ΔURA.

[0115] The culture temperature used by Yarrowia lipolytica is 28°C, and the shaker speed is 220rpm during liquid culture, and the culture medium used is as follows:

[0116] YPD liquid medium: glucose 20g / L, peptone 20g / L, yeast extract 10g / L, if solid medium is required, add 1.5% a...

Embodiment 2

[0190] Embodiment 2: fermenter fermentation of recombinant bacterial strain

[0191] 1. Materials and methods

[0192] 1.1 Fermentation medium

[0193] Seed culture medium (YNB medium): YNB 3g / L, glucose 60g / L, ammonium sulfate 35g / L, YNB and ammonium sulfate were sterilized and added by filtration.

[0194] Fermentation medium (FM medium): glucose 50g / L, ammonium sulfate 35g / L, yeast powder 38g / L, potassium dihydrogen phosphate 8g / L, disodium hydrogen phosphate dodecahydrate 6g / L, magnesium sulfate heptahydrate 5g / L, defoamer 1mL / L.

[0195] Feed control: control according to the residual sugar concentration in the fermenter, and use 600g / L glucose feed after sterilization.

[0196] 1.2 Fermentation process of 50L fermenter

[0197] The preserved strains were taken out from -80°C and thawed on ice. Draw 100μl and inoculate into 250mL YNB medium, culture on a shaker at 220rpm and 28°C for 24 hours, then transfer to a 5L seed fermentation tank with an OD of about 0.01 after...

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Abstract

The invention provides an application of a fatty acid elongase gene in yeast synthesis of nervonic acid. Specifically, the invention provides an engineering bacterium for producing nervonic acid, a gene expression cassette is integrated in a genome of the engineering bacterium, and the gene expression cassette expresses a protein coded by a 3-ketoester acyl-CoA synthase (KCS) gene. The engineering bacterium disclosed by the invention can obviously improve the yield of nervonic acid.

Description

technical field [0001] The invention relates to the field of biotechnology, and more specifically relates to the application of a fatty acid elongase gene in the synthesis of nervonic acid by yeast. Background technique [0002] Neural acid (cis-15-Tetracosenoic acid, 24:1Δ15) is a very long chain monounsaturated fatty acid (VLCMFAs), mainly in the white matter of animal brain in the form of glycosphingolipids and sphingomyelin and myelin in nerve fibers and is an important component of biofilms. Neural acid plays an important role in medicine and health care. It has the functions of promoting brain development, improving memory, regulating blood lipids, and enhancing immunity. It can be used to treat neurological disorders such as multiple sclerosis. In addition, studies have shown that nervonic acid can promote the development of the nervous system, especially in the growth and development of infant brain nerve cells and optic nerve cells. [0003] At present, the synthe...

Claims

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Application Information

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IPC IPC(8): C12N1/19C12N15/54C12P7/6409C12R1/645
CPCC12N9/1029C12P7/6409C12N9/10C12R2001/645C12R2001/085C12N1/16C12P7/6436C12P7/6454
Inventor 王士安韩兴丰阮建昌苏航史鹏辉樊伟明李福利
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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