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Macleaya cordata protopine-6-hydroxylase gene optimization sequence and application thereof

A hydroxylase and gene optimization technology, applied in the field of genetic engineering and bioengineering, can solve the problems of unreported P6H gene expression and achieve the effects of improving enzyme catalytic efficiency, reducing production cost and increasing content

Active Publication Date: 2019-04-12
MICOLTA BIORESOURCE INC LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, no scholars have reported the expression of P6H gene in different species

Method used

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  • Macleaya cordata protopine-6-hydroxylase gene optimization sequence and application thereof
  • Macleaya cordata protopine-6-hydroxylase gene optimization sequence and application thereof
  • Macleaya cordata protopine-6-hydroxylase gene optimization sequence and application thereof

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Embodiment Construction

[0030] In order to better illustrate the content of the invention, the present invention will be further verified by specific examples below. It is specially stated here that the embodiments are only for describing the present invention more directly, they are only a part of the present invention, and cannot constitute any limitation to the present invention.

[0031] as attached figure 1 The synthetic pathways of sanguinarine and chelerythrine in Boluohui shown, protropine-6-hydroxylase (P6H) is involved in the biogenesis of sanguinarine (SAN) and chelerythrine (CHE) Synthetic steps, which can catalyze allocryptine (ALL) to generate dihydrochelerythrine (DHCHE) and catalyze proopiate (PRO) to generate dihydrosanginarine (DHSAN). Since P6H belongs to cytochrome P450 oxidoreductase, as a monooxygenase, it needs to be co-expressed with cytochrome P450 reductase (CPR) to play a role in protein catalysis, and CPR plays the role of transferring electrons in the reaction.

[0032]...

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Abstract

The present invention provides a macleaya cordata protopine-6-hydroxylase gene optimization sequence and an application thereof, and specifically discloses an optimization sequence MC11229opt obtainedby codon optimization of a protopine-6-hydroxylase gene MC11229, a recombinant expression vector containing the optimization sequence MC11229opt, an engineered yeast of the recombinant expression vector containing the optimization sequence MC11229opt, and applications thereof in preparing dihydrosanguinarine / sanguinarine. An optimization sequence obtained by codon optimization of a functional gene MC11229 of macleaya cordata participating in synthesizing the sanguinarine and chelerythrine according to yeast preferred codons is integrated into saccharomyces cerevisiae for heterologous expression to realize microbial transformation of the sanguinarine and chelerythrine, and compared with non-optimization functional genes, the optimization sequence can greatly improve enzyme catalytic efficiency, improves content of catalytic products of the dihydrosanguinarine and sanguinarine, and reduces production costs of the dihydrosanguinarine and sanguinarine.

Description

technical field [0001] The invention relates to an optimized sequence of boluohuiprotropine-6-hydroxylase gene and its application, belonging to the technical field of genetic engineering and bioengineering. technical background [0002] Medicinal plants are plants that have the curative effect of preventing and curing diseases in Chinese traditional medicine and are used as the source of medicines. They have a very important position in Chinese medicine. The active ingredients of many medicinal plants are their secondary metabolites, which have attracted widespread attention due to their unique curative effect and low toxicity and side effects. my country is rich in wild medicinal plant resources, but due to the destruction of the natural environment and the over-exploitation and abuse of medicinal plants in recent years, the medicinal plant resources in my country are on the verge of exhaustion. Wild resources can no longer meet people's needs, and there are problems such...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N9/02C12N15/81C12N1/19C12P17/18C12R1/865
CPCC12N9/0073C12N15/81C12N2800/22C12P17/18C12Y114/13055
Inventor 黄鹏曾建国
Owner MICOLTA BIORESOURCE INC LTD
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