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Method for improving yield of homoorientin produced by recombinant bacteria through coordinated regulation strategy

A technology of isoorientin and recombinant bacteria, applied in the fields of genetic engineering technology and microorganisms, can solve the problems such as the inability to greatly increase the yield, and achieve the effects of inhibiting the accumulation of acetic acid, increasing the yield and simple process

Active Publication Date: 2019-03-01
NANJING FORESTRY UNIV
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  • Claims
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AI Technical Summary

Problems solved by technology

Defects in any aspect will lead to the inability to increase the yield significantly

Method used

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  • Method for improving yield of homoorientin produced by recombinant bacteria through coordinated regulation strategy
  • Method for improving yield of homoorientin produced by recombinant bacteria through coordinated regulation strategy
  • Method for improving yield of homoorientin produced by recombinant bacteria through coordinated regulation strategy

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Embodiment 1

[0021] 1. Construction of recombinant bacteria BL21-CGT-I

[0022] The carboside glycosyltransferase Gt6CGT gene (AB985754.1) derived from Gentiana triflora was optimized according to the dominant codon table of Escherichia coli K12 (http: / / www.kazusa.or.jp / codon / ), and the amino acid The codons with high frequency of sense codons are used as dominant codons, and finally the optimized sequence information of the carboside glycosyltransferase gene is obtained, which is synthesized by Shanghai Jierui Bioengineering Co., Ltd. and added to the N-terminus and C-terminus of the gene respectively The Nco I and BamH I restriction sites were used to obtain the optimized Gt6CGT gene, the gene sequence of which is shown in SEQ ID NO:1.

[0023] The optimized Gt6CGT gene was double-digested with Nco I and BamH I, and the recombinant plasmid pCDFDuet-1 was also double-digested with Nco I and BamH I, and the rubber was harvested separately, concentrated and ligated overnight at 16°C, and th...

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Abstract

The invention provides a method for improving the yield of homoorientin produced by recombinant bacteria through a coordinated regulation strategy. Based on the recombinant bacteria carrying a C-glycosides glycosyl transferase Gt6CGT gene, the yield of the homoorientin produced by the recombinant bacteria is greatly improved through regulation of supply of a UDP-glucose by introducing genes, optimization of an induction conversion strategy and control of acetic acid accumulation. Through the coordinated regulation strategy, the recombinant bacteria BL21-CGT-III homoorientin yield reaches 1371mg / L, the molar conversion rate of a substrate reaches 97.4% and is 40 times the current production peak, and the process for preparing the homoorientin is simple.

Description

technical field [0001] The invention belongs to the field of genetic engineering technology and microorganisms, and specifically relates to a method for improving the yield of isoorientin produced by recombinant bacteria by using a coordinated regulation strategy. Background technique [0002] Carbon glycoside flavonoids are flavonoids whose sugar groups are directly connected to flavonoids through C-C bonds. They are widely found in wheat, corn, rice, mung beans, blueberries, lemons, black tea and bamboo leaf extracts. Compared with oxyglycoside flavones, carbon glycoside flavones have the characteristics of stable structure and ability to penetrate deep into the lesion. Isoorientin, also known as luteolin 6-C-glucoside, is a luteolin glycoside carbon glycoside flavonoid compound, and is one of the main components of flavonoids in bamboo leaf extract. Studies in recent years have found that isorientin can scavenge free radicals, resist the loss of cell viability caused by ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/70C12N15/54C12P17/16
CPCC12N9/1048C12N9/1051C12N9/1241C12N15/70C12P17/162C12Y204/0102C12Y207/07009
Inventor 赵林果孙晴裴建军
Owner NANJING FORESTRY UNIV
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