Hansenula polymorpha recombinant strain and application thereof in biosynthesis of paclitaxel

A technology of Hansenula polymorpha and recombinant strains, applied in the direction of microorganism-based methods, microorganisms, biochemical equipment and methods, etc., can solve the problems that paclitaxel has not been reported, and achieve the effect of easy high-density fermentation and easy separation and extraction

Active Publication Date: 2014-07-16
SHAANXI UNIV OF SCI & TECH
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Problems solved by technology

[0007] At present, there are no foreign reports on the use of low-energy ion implantation to mediate medicinal plant genomic DNA transformation yeast technology to

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  • Hansenula polymorpha recombinant strain and application thereof in biosynthesis of paclitaxel
  • Hansenula polymorpha recombinant strain and application thereof in biosynthesis of paclitaxel
  • Hansenula polymorpha recombinant strain and application thereof in biosynthesis of paclitaxel

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Embodiment Construction

[0026] The present invention will be described in detail below in conjunction with the accompanying drawings and embodiments.

[0027] The present invention uses H.polymorpha DL-1 as the starting strain (the source is ATCC No.26012), utilizes low-energy ion irradiation injection to mediate the transformation of Taxus chinensis genomic DNA, and obtains paclitaxel-producing recombinant strains by screening, and genetically Stability and corresponding fermentation tests.

[0028] (1) culture medium

[0029] The composition of liquid inorganic medium: glycerol 19g, (NH 4 ) 2 SO 4 6g,K 2 HPO 4 0.24g, MgSO 4 ·7H 2 O1.4g, CaCl 2 0.38g, NaCl0.6g, NaNO 3 0.58g, 0.2mL iron salt solution, 1mL thiamine hydrochloride solution, 1mL PTM trace element solution, dilute to 1L with water, pH 6.5;

[0030] Wherein each solution batching is as follows:

[0031] Iron salt solution: FeCl 3 ·6H 2 O30g, dissolved in water and dilute to 1L;

[0032] Thiamine hydrochloride solution: thiami...

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Abstract

The invention provides a hansenula polymorpha recombinant strain and an application thereof in biosynthesis of paclitaxel. A total DNA transformation yeast of mediated taxus chinensis genome is injected to obtain a hansenula polymorpha recombinant strain CGMCC No.8999 for producing paclitaxel by using low-energy ions. The technical breakthrough for biological fermentation to produce paclitaxel by using the recombined yeast is achieved, and a new way is provided for solving paclitaxel source shortage, protecting taxus chinensis plant resources in China and sustainable development thereof.

Description

technical field [0001] The invention belongs to the field of transgenic yeast recombinant bacteria producing paclitaxel obtained through genetic transformation, and specifically relates to a Hansenula polymorpha recombinant strain obtained by genetic transformation of the total genome DNA of Taxus chinensis in yeast by using low-energy ion implantation and its biosynthesis of paclitaxel in the application. Background technique [0002] Paclitaxel (trade name Taxo1) is a natural anticancer drug with broad-spectrum and definite efficacy recognized in the world today. Its molecular structure is derived from baccatin III and phenylalanine linked to the 13th carbon. tricyclic diterpenoid alkaloids. Paclitaxel is the only drug known to promote microtubule polymerization and stabilize the polymerized microtubules. After the cells are exposed to paclitaxel, a large number of microtubules accumulate in the cells, and the accumulation of these microtubules interferes with various cel...

Claims

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Application Information

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IPC IPC(8): C12N1/19C12P17/02C12R1/78
Inventor 钱卫东赵德志蔡长龙毛培宏王婷陈雪峰施春阳
Owner SHAANXI UNIV OF SCI & TECH
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