Genetic engineering strain taking tyrosine as substrate to synthesize naringenin and construction method thereof

Abstract

The invention discloses a genetic engineering stain taking tyrosine as a substrate to synthesize naringenin and a construction method thereof, belonging to the field of synthetic biology or metabolic engineering. According to the construction method disclosed by the invention, a synthesis route constituted by six genes, namely a gene matB encoding a malonic acid transporting enzyme, a gene matC encoding a malonic acid absorption route, a gene encoding a tyrosine ammonialyase (TAL), the gene encoding a 4-cinnamic acid: coenzyme A ligase (4CL), the gene encoding a chalcone synthase (CHS) and the gene encoding a chalcone isomerase (CHI) is introduced into Escherichia coli BL21 to obtain a recombinant strain capable of taking the tyrosine as the substrate to synthesize the naringenin, the synthesis route is further optimized through a modular transformation theory, the strain is finally fermented in a shaking flask for 72 hours by taking the tyrosine as the substrate, and the yield of the naringenin achieves 90mg / L. A strategy adopted by the construction method disclosed by the invention provides certain reference significance for future production of natural small molecular substances by a microbiological method.

Description

technical field [0001] The invention relates to a genetically engineered bacterium for synthesizing naringenin with tyrosine as a substrate and a construction method thereof, belonging to the field of synthetic biology or metabolic engineering. 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 for flavonoids has grown by more than 30% every year, and has a broad market in the field of pharmaceuticals and nutritional chemicals. application prospects. [0003] Flav...

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Problems solved by technology

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

At present, domestic and foreign studies on the production of naringenin by microbial fermentation are all using coumaric acid as a substrate. However, coumaric acid is expensive and has low solubility, so it is difficult to exist in general culture media; in addition, the current fermentation method Both adopt a two-step method of production, 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 other methods and placed in a basic medium for fermentation. Obviously, these will inhibit The application of microbial method in large-scale industrial production

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