Method for efficient enzyme catalytic synthesis of sanguinarine and chelerythrine

Abstract

The invention discloses a method for efficient enzyme catalytic synthesis of sanguinarine and chelerythrine. The method comprises the following steps: respectively screening optimal genes with high expression efficiency from a known protopine-6-hydroxylase gene, a dihydrobenzophen anthridine oxidase gene and a cytochrome P450 reductase gene through heterologous expression and result comparison andanalysis, and carrying out codon optimization on selected optimal genes; establishing optimal gene sequences on expression carriers, transferring into a yeast engineering bacterium, and carrying outtransformation so as to obtain a recombinant yeast engineering strain; finally feeding the recombinant yeast engineering strain with a macleaya cordata leaf raw material liquid precursor to carry outfermentation, thereby obtaining sanguinarine and chelerythrine. By adopting the method, the enzyme catalysis efficiency of the sanguinarine and the chelerythrine is improved from multiple aspects suchas gene levels and fermentation processes, fumarine and allocryptopine which are high in alkaloid content in leaves can be transformed into sanguinarine and chelerythrine with high values, and comprehensive utilization of macleaya cordata resources can be achieved.

Description

technical field [0001] The invention relates to the technical field of enzyme-catalyzed synthesis of sanguinarine and chelerythrine, in particular to a method for efficiently enzymatically synthesizing sanguinarine and chelerythrine. technical background [0002] Macleaya cordata (Willd.) R.Br. belongs to the genus Macleaya cordata (Willd.) R.Br., and is also known as trumpet pole, Luohui, mountain trumpet, mountain phoenix tree, three money three, etc. It grows in hills and low mountains , forest edge, grassland, roadside, is a kind of wild herbaceous plant. Boluohui is mainly distributed in China, East Asia, North America and Europe. The plants of the genus Boluohui include two species of Boluohui and M.microcarpa (Maxim) Fedde. As a traditional Chinese herbal medicine, it was first seen in "Supplements to Materia Medica", and it is used as a kind of maggot-killing green herbal medicine in the folk. widely used. With the continuous deepening of research, it was found th...

AI Technical Summary

Problems solved by technology

At present, most of the extracts of Boluohui come from the fruit pods of wild resources, the source is limited, and the content of sanguinarine is low, which makes the price of sanguinarine expensive, which limits the development of its industry

On the other hand, the precursor substances, which account for a large proportion of the total alkaloids, have not been completely transformed, and are treated as waste in the extraction process without comprehensive and efficient utilization, resulting in a great waste of

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