Fusion protein capable of improving the conversion efficiency of dammarenediol and its construction method

A technology for fusion protein and dammarene diol, which is applied in the field of fusion protein and construction, can solve the problem of low conversion efficiency of dammarene diol, and achieve the effect of being conducive to efficient synthesis and improving conversion efficiency
CN104450769BActive Publication Date: 2017-08-08TIANJIN UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
TIANJIN UNIV
Publication Date
2017-08-08

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Abstract

The invention discloses a fusion protein capable of increasing the conversion efficiency of dammarendiol and a construction method. The construction method comprises the following steps: (1) excising the front 138 basic groups of the end 5' of the cytochrome-NADPH-reductase 1 gene AtCPR1 in arabidopsis to obtain a sequence shown in SEQ ID NO. 2; (2) removing the termination codon TAA of the end 3' of the synthase gene PPDS of protopanoxadiol in ginseng shown in SEQ ID NO. 3, and connecting with a sequence at the end 5' of the sequence shown in SEQ ID NO. 2 to construct a genetic element of the fusion protein; (3) connecting the genetic element of the fusion protein with the internal promoter and the terminator of a saccharomyces cerevisiae cell to construct a genetic expression kit of the fusion protein, and transforming to enter the saccharomyces cerevisiae cell for expression. The fusion protein constructed by the method disclosed by the invention is capable of increasing the conversion efficiency of converting from dammarendiol to protopanoxadiol.
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Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a fusion protein capable of improving the conversion efficiency of dammarenediol and a construction method. Background technique

[0002] Ginsenoside is the main active ingredient of traditional Chinese medicine ginseng, which has the pharmacological effects of protecting cardiovascular, anti-fatigue, anti-aging and anti-cancer. Traditional ginseng cultivation, tissue culture and downstream plant tissue extraction have been difficult to meet the market demand for ginsenosides.

[0003] As a triterpenoid, the endogenous metabolic pathway of yeast can provide the precursor 2,3-oxysqualene for the synthesis of ginsenosides. Subsequent epoxidation, oxidation, and glycosyl addition are accomplished by dammarenediol synthase, protopanaxadiol synthase, and glycosyltransferase, respectively. The dammarenediol synthase gene was discovered in 2006. Tansakul et al. reported that PNA (DDBJ da...

Claims

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