Applications of glycosyltransferase and related materials thereof in construction of engineering bacteria for producing ginsenoside Rb1 and Rg1

Abstract

The invention discloses applications of glycosyltransferase and related materials thereof in the construction of engineering bacteria for producing ginsenoside Rb1 and Rg1. A glycosyltransferase genePn3-32 which can catalyze ginsenoside Rd to generate ginsenoside Rb1 can be successfully identified through a synthetic biological method; and the gene can simultaneously catalyze ginsenoside F1 to generate ginsenoside Rg1 and construct recombinant yeast producing the ginsenoside Rb1 and the ginsenoside Rg1. Through experiments, the constructed recombinant yeast producing the ginsenoside Rb1 and the ginsenoside Rg1 can simultaneously generate the ginsenoside Rb1 and the ginsenoside Rg1. Pn1-31, Pn-3-29, Pn3-31 and Pn3-32 glycosyltransferase genes in medicinal plant radix notoginseng are firstly utilized to continuously catalyze protopanaxadiol and protopanaxatriol to synthetize the ginsenoside Rb1, the ginsenoside Rg1 and corresponding intermediates, so that novel cases can be provided formicrobial strains to produce natural products.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the application of glycosyltransferase and related materials in constructing engineering bacteria producing ginsenosides Rb1 and Rg1. Background technique [0002] Ginsenoside Rb1 (Ginsenoside Rb1), Ginsenoside Rd (Ginsenoside Rd) and Ginsenoside Rg1 (Ginsenoside Rg1) are all dammarane-type triterpene saponins, mainly distributed in Araliaceae plants such as ginseng, Panax notoginseng, American ginseng, etc. Among them, it has a variety of pharmacological activities and is a chemical substance of great medicinal value. Among them, ginsenoside Rb1 has improved regulatory effects on the central nervous system, cardiovascular system, immune system and anti-tumor. Ginsenoside Rd has significant biological activity in cardiovascular system, immune system, renal function protection and anti-tumor effect. Ginsenoside Rg1 has a natural anti-oxidation effect, affects the nervous ...

AI Technical Summary

Problems solved by technology

At present, relevant progress has been made in the research on the biosynthetic pathway of ginsenosides, including some key enzymes in the synthetic pathway of ginsenosides, such as squalene synthase (Squalene synthase, which catalyzes the isoprenoid pathway towards sterols and triterpene saponins). SS), squalene epoxidase (Squaleneepoxidase, SE) that catalyzes the generation of 2,3-oxidized squalene, dammarene diol synthase (DS) that catalyzes the generation of dammarene diol, and responsible for hydroxylation Cytochrome P450 enzymes, etc., but the biosynthesis of various glycosylated saponins in Panax notoginseng is relatively complicated, and has not yet been fully explored

In the study of the effect of calcium ions on the biosynthesis of Panax notoginseng and ginsenoside Rb1, Zhong Jianjiang et al. found that the crude enzyme solution extracted from Panax notoginseng plant tissue cells had the activity of catalyzing the production of ginsenoside Rd from ginsenoside Rb1, but no related glycosyltransferases were found. Genes (Yue CJ et al., Biotechnol Bioeng, 2005, 89(4):444-452)

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