Escherichia coli engineering bacterium for synthesizing pinocembrin by using glucose as substrate and application thereof

Abstract

The invention discloses an Escherichia coli engineering bacterium for synthesizing pinocembrin by using glucose as a substrate and application thereof and belongs to the field of genetic engineering or metabolic engineering. Synthetic biology and metabolic engineering means are adopted to respectively synthesize a synthesis route consisting of eight genes in pinocembrin metabolism to be introduced into Escherichia coli (BL21), accordingly the recombinant bacterium for synthesizing the flavone framework material pinocembrin by using the glucose as the substrate, and the pinocembrin yield reaches to 21mg / L after glucose serves as the unique substrate in a shake flask to be fermented for 60 hours. The synthesis route consisting of the eight genes is successfully built in the Escherichia coli, and synthesis from the glucose into the pinocembrin is achieved successfully. Certain reference significance is provided for flavonoid material production by means of a microbiological method in the future.

Description

technical field [0001] The present invention relates to a kind of Escherichia coli engineering bacterium producing pinesin, which is an overexpressed aroF and pheA by means of synthetic biology and metabolic engineering fbr , PAL, 4CL, CHS, CHI, matB, matC and other eight gene synthesis pathways to realize the Escherichia coli engineering bacteria that use glucose as a substrate to produce pinocin. Background technique [0002] Flavonoids (flavonoids) are a class of compounds that exist in nature and have a 2-phenylchromone structure. Most of their hydroxy derivatives are yellow, hence the name. Flavonoids have a variety of biological activities, mainly including cardiovascular system, antibacterial and antiviral, antitumor, antioxidative free radicals, anti-inflammation, etc. Prevention, treatment or auxiliary treatment effects, and the human body cannot synthesize flavonoids and can only be obtained from plant foods. Therefore, in recent years, the market of flavonoids ha...

AI Technical Summary

Problems solved by technology

[0004] At present, the industrial production of pinocin mainly adopts methods such as plant extraction and chemical synthesis, but how to obtain the required high-purity flavonoids from the complex extracts of plants is still a technical problem that cannot be solved by human beings. Factors such as harmful chemical reagents and the complex structure of flavonoids limit the use of chemical methods. For this reason, microbial fermentation is favored by the use of cheap and non-polluting raw materials, low pollution emissions, and low energy requirements. more and more attention

At present, domestic and foreign studies on the production of pinecone by microbial fermentation all need to add expensive precursor substances such as phenylalanine or cinnamic acid to the fermentation broth, and from the perspective of food safety, since cinnamic acid is derived from petroleum It is synthesized by industrial chemical synthesis, so the addition of cinnamic acid in particular cannot be used commercially

In addition, the current fermentation method is produced by a two-step method, that is, the strain first grows in a nutrient-rich medium, and when a certain bacterial concentration is reached, all the bacteria are collected by centrifugation and placed in a basic medium for fermentation , obviously this fermentation method cannot be applied to large-scale industrialization