High-efficiency enzyme-catalyzed method for synthesizing sanguinarine and chelerythrine

Abstract

A highly efficient method for catalyzing the synthesis of sanguinarine and chelerythrine, comprising: from the known protopine-6-hydroxylase gene, dihydrobenzophenanthridine oxidase gene and cytochrome P450 reductase gene, by means of heterologous expression and result comparison and analysis, screening out an optimal gene having high expression efficiency, then performing codon optimization on the selected optimal gene; constructing the optimal gene sequence on an expression vector, then transferring same into yeast engineering bacteria for transformation so as to obtain recombinant yeast engineering strains; finally, feeding the recombinant yeast engineering bacteria by using a leaf raw material liquid precursor of Macleaya cordata for fermentation, so as to obtain a product. The invention improves the enzyme catalytic efficiency of sanguinarine and chelerythrine in various aspects such as gene level and fermentation process; the leaf raw material liquid, which is not a traditional medicinal part of Macleaya cordata, is directly used for fermentation with the engineering bacteria, and protopine and allocryptine having high alkaloid content in leaves are converted into high-value sanguinarine and chelerythrine to achieve the comprehensive utilization of Macleaya cordata resources.

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. [0002] technical background [0003] 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 ...

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 resources

Since every 0.1% increase in the content of sanguinarine in the plant can reduce the extraction cost by 10%, while traditional cultivation and breeding require a long cycle for improvement, are greatly affected by the environment, have low efficiency, are difficult to plant on a large scale, and have limited potential for improvement. The effective conversion of precursors to final products by engineering bacteria constructed in vitro by modern molecular biology techniques is a new source of sanguinarine

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