Genetic engineering bacterium for producing glycollic acid by acetic acid, and building method and application thereof

A technology of glycolic acid and engineering bacteria, applied in the field of genetic engineering, fermentation engineering, and biotechnology, can solve the problem of high cost of raw materials and achieve good industrial application prospects

Active Publication Date: 2018-01-19
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, studies have reported glycolic acid synthesis pathways and production strains using gluc

Method used

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  • Genetic engineering bacterium for producing glycollic acid by acetic acid, and building method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036]Example 1. Construction of recombinant bacteria E.coliΔGA4 (pUC19-gltA-acs-pta-ackA+pMCS-ycdW-aceAK)

[0037] 1. Construction of recombinant expression vector pUC19-gltA-acs-pta-ackA

[0038] 1. The artificially synthesized DNA shown in Sequence 1 in the sequence table contains the gltA gene, the upstream is the SacI site, and the downstream is the KpnI site, wherein the 9th to 51st nucleotides are the promoter sequence, and the 70th to 1353rd nucleotides are the promoter sequence. Nucleotide is gltA gene sequence.

[0039] 2. The DNA shown in Sequence 2 in the sequence list is artificially synthesized, containing the acs gene, with the KpnI site upstream and the XbaI site downstream, wherein the 9th-51st nucleotide is the promoter sequence, and the 70th-2028th nuclear Nucleotide is the sequence of acs gene.

[0040] 3. The artificially synthesized DNA shown in sequence 3 in the sequence list contains the pta-ackA gene, the upstream is the XbaI site, and the downstream...

Embodiment 2

[0124] Example 2, the application of recombinant bacteria E.coliΔGA4 (pUC19-gltA-acs-pta-ackA+pMCS-ycdW-aceAK) in the production of glycolic acid

[0125] 1. Shake flask culture experiment of recombinant bacteria E.coliΔGA4 (pUC19-gltA-acs-pta-ackA+pMCS-ycdW-aceAK)

[0126] 1. Put the E.coliΔGA4 (pUC19-gltA-acs-pta-ackA+pMCS-ycdW-aceAK) prepared in Example 1 in LB liquid medium containing ampicillin and kanamycin at 37°C, Cultivate for 16 hours at a rotational speed of 200 rpm, and use it as a seed solution.

[0127] 2. The seed solution was inoculated into the MM liquid medium according to the inoculation amount of 4% by volume, and the amount of liquid in the 500ml shake flask was 50ml, and cultivated for 48h at 37°C and 200rpm at a rotating speed, during which the fermentation broth was collected.

[0128] 3. Quantitative detection of glycolic acid produced by genetically engineered bacteria and acetic acid consumed by high performance liquid chromatography. The specific ...

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Abstract

The invention discloses a genetic engineering bacterium for producing glycollic acid by acetic acid, and a building method and application thereof. A method for preparing the genetic engineering bacterium for producing glycollic acid provided by the invention comprises the following steps of improving the expression and/or activity of acetylcoenzyme A synthetase, phosphotransacetylase, acetokinase, citrate synthase, isocitrate lyase, isocitrate dehydrogenase kinase and glyoxylate reductase in the recipient bacterium; reducing the expression and/or the activity of malate synthase, glycolate oxidase and isocitrate lyase repressor protein in the recipient bacterium, so that the engineering bacterium for producing the glycollic acid is obtained. The recipient bacterium is a bacterium capable of using acetic acid as a carbon source for growth. The prepared genetic engineering bacterium uses acetic acid as the carbon source for producing the glycollic acid; in addition, the production quantity and yield of the glycollic acid in shaking culture can reach the relatively high level; relatively good industrial application prospects are realized.

Description

technical field [0001] The invention belongs to the fields of biotechnology, genetic engineering and fermentation engineering, and relates to a genetically engineered bacterium for producing glycolic acid from acetic acid, a construction method and application thereof. Background technique [0002] Glycolic acid, also known as glycolic acid, is the simplest member of the hydroxycarboxylic acid family. Glycolic acid is a small molecule with both the functions of alcohol and moderately strong acid, and it is an important raw material for organic synthesis. Glycolic acid is widely used in many fields such as chemical industry, medicine, textile and metallurgy. For example, the mixed acid of 2% glycolic acid and 1% formic acid is an efficient and low-cost cleaning agent, which can be used for cleaning boilers, pipes, condensers and heat exchangers; glycolic acid has small molecules and can effectively penetrate pores to solve Skin aging, wrinkles, acne and other problems are o...

Claims

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Application Information

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IPC IPC(8): C12N15/70C12N1/21C12P7/42C12R1/19
Inventor 李正军陈静李良康笪央央李微
Owner BEIJING UNIV OF CHEM TECH
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