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Betulinic acid biosynthetic pathway screening method based on hansenula polymorpha

A technology of Hansenula polymorpha and betulinic acid, applied in the field of genetic transformation and ultra-distant genetic transformation, to achieve the effect of easy extraction and separation

Active Publication Date: 2018-11-27
SHAANXI UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
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Problems solved by technology

However, there have been no reports at home and abroad on the use of low-energy ion implantation-mediated ultra-distant genetic transformation to transform yeast biosynthesis of betulinic acid

Method used

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Experimental program
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Embodiment Construction

[0019] The present invention is described in detail below in conjunction with embodiment.

[0020] The present invention takes H.polymorpha DL-1 as the starting strain (the source is ATCC No.26012), uses low-energy ion irradiation injection to mediate the random genetic transformation of birch genome DNA into Hansenula polymorpha, and obtains birch-producing polymorpha through qualitative and quantitative screening. Recombinant yeast strains of fatty acids, and the genetic stability and corresponding fermentation tests were carried out, so as to provide genetic materials for the efficient analysis of betulinic acid biosynthesis pathway.

[0021] (1) culture medium

[0022] The composition of YPD liquid medium is: glucose 20.0g / L, peptone 20.0g / L, yeast extract 10.0g / L, pH 6.5;

[0023] The composition of YPD solid medium is: glucose 20.0g / L, peptone 20.0g / L, yeast extract 10.0g / L, agar 20g / L, pH 6.5.

[0024] (2) The method on which the screening is based

[0025] (1) Quali...

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Abstract

The invention provides a betulinic acid biosynthetic pathway screening method based on hansenula polymorpha. By primarily injecting low-energy ions into mediated silver birch genomic DNA to convert saccharomycetes, a material is provided for efficiently analyzing a betulinic acid biosynthetic pathway with reverse thinking, thereby providing a novel path for solving shortage of sources of betulinicacid, and protecting sources of fallen leaves magaphanerophytes silver birches and sustainable development thereof.

Description

technical field [0001] The invention belongs to the technical field of ultra-distant genetic transformation, and in particular relates to the genetic transformation of birch genome DNA in yeast mediated by ion implantation. Background technique [0002] Betulinic acid (BA) is a pentacyclic triterpene acid compound, which exists in plants such as white birch (Betulapubescens), Syzygium formosanum, and Ziziphus mauritiana. Studies in recent years have shown that BA has many pharmacological activities, such as anti-tumor, anti-inflammatory, anti-bacterial, anti-HIV, anti-malarial, etc., especially in anti-tumor, such as melanoma. Because BA has many pharmacological activities, coupled with its high safety, the American Institute of Cancer Research has included the development of betulinic acid in the rapid research and development program, which has attracted much attention to BA. [0003] At present, BA is mainly extracted from birch bark, but the content of BA in birch is ve...

Claims

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Application Information

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IPC IPC(8): C12N15/87C12P33/00C12R1/78
CPCC12N15/87C12P33/00
Inventor 钱卫东卢家兴段冉冉
Owner SHAANXI UNIV OF SCI & TECH
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