Trifunctional enzyme for de novo synthesis of flavanone as well as synthesis method and application of trifunctional enzyme

Abstract

The invention discloses a trifunctional enzyme for de novo synthesis of flavanone, wherein 4-coumaroyl-coenzyme A ligase genes, chalcone isomerase genes and chalcone synthase genes are cloned to a prokaryotic expression vector to construct recombinant plasmids, then escherichia coli is converted to induce expression of recombinase; and then purified recombinase is utilized to establish a cell-free synthesis system, and in-vitro enzymatic synthesis of flavanone is realized. According to the invention, the problems of low yield and low substrate conversion rate in an in-vitro multi-enzyme synthesis system of flavanone are solved, and an in-vitro enzymatic synthesis system with simple components is established at the same time. The system does not have a complex regulation effect in engineering bacteria cells, the reaction process is easy to accurately control, byproducts are few, product separation is relatively simple, the production period is obviously shortened, and the production cost is obviously reduced.

Description

technical field [0001] The invention relates to a trifunctional enzyme for de novo synthesis of flavanones, its synthesis method and application, the classification number is C12P, and it belongs to the technical field of biomedicine. Background technique [0002] Flavonoids generally refer to a large class of natural small molecule plant secondary metabolites whose basic core is 2-phenylchromone, and now generally refer to two benzene rings connected to each other through the central three-carbon chain to form a C6-C3-C6 A series of compounds with the main structure. According to the difference of the central C ring, flavonoids are generally divided into six subgroups: flavanols, flavonoids, flavonols, flavanones, anthocyanins and isoflavones. These compounds have a variety of biological and pharmacological activities, and are currently one of the hot spots in the field of biomedicine, and have broad application and development prospects in medicine, food, cosmetics and ot...

AI Technical Summary

Problems solved by technology

However, as far as the in vitro enzymatic synthesis of complex compounds is concerned, the actual production process is cumbersome, and the formation of substrate channels is not easy, which often leads to low yields of in vitro enzymatic synthesis

Images