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Optimized sequence of macleaya cordata dihydrobenzophenanthridine oxidase gene and application thereof

A technology for dihydrotriphenanthridine oxidase and gene optimization, which is applied in the fields of genetic engineering and bioengineering, can solve problems such as reports on the expression of non-P6H genes, and achieve the effects of improving enzyme catalysis efficiency, reducing production costs, and increasing content

Active Publication Date: 2019-04-12
MICOLTA BIORESOURCE INC LTD
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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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  • Optimized sequence of macleaya cordata dihydrobenzophenanthridine oxidase gene and application thereof
  • Optimized sequence of macleaya cordata dihydrobenzophenanthridine oxidase gene and application thereof
  • Optimized sequence of macleaya cordata dihydrobenzophenanthridine oxidase gene 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, the dihydrobenzophenanthridine oxidase (DBOX) gene is involved in the biogenesis of sanguinarine (SAN) and chelerythrine (CHE) Synthetic steps, which can catalyze dihydrochelerythrine (DHCHE) to generate chelerythrine (CHE) and catalyze dihydrosanginarine (DHSAN) to generate sanguinarine (SAN). Dihydrotriphenylene oxidase (DBOX) belongs to the flavoprotein oxidase gene.

[0032] In the applicant's previous research, the flavin protein oxidase-related gene sequence of other species (poppy, poppy) that had been publis...

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Abstract

The invention provides an optimized sequence of a macleaya cordata dihydrobenzophenanthridine oxidase gene and application thereof. The invention comprises codon optimization on a dihydrobenzophenanthridine oxidase gene MC6408 to obtain an optimized sequence MC6408opt, a recombinant expression vector of the optimized sequence MC6408opt, yeast engineering bacteria containing the recombinant expression vector of the optimized sequence MC6408opt, and application of the optimized sequence MC6408opt, recombinant expression vector and yeast engineering bacteria in the preparation of pseudochelerythrine. The functional gene MC6408 of macleaya cordata participating in synthesis of pseudochelerythrine and chelerythrine is subjected to codon optimization according to a yeast preferred codon to obtain the optimized sequence; and the optimized sequence is integrated into saccharomyces cerevisiae for heterologous expression, so that microbial transformation of the pseudochelerythrine and chelerythrine is realized. Compared with an unoptimized functional gene, the optimized gene can greatly improve the enzymatic efficiency of the dihydrobenzophenanthridine oxidase, increase the content of the catalytic product pseudochelerythrine, and reduce the production cost of the pseudochelerythrine.

Description

technical field [0001] The invention relates to an optimized gene sequence of Boluohui dihydrotriphenylene oxidase 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 ...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N9/06C12N15/81C12N1/19C12P17/18C12R1/865
CPCC12N9/0032C12N15/81C12N2800/22C12P17/18C12Y105/03012
Inventor 黄鹏曾建国
Owner MICOLTA BIORESOURCE INC LTD
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