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Paclitaxel genome rearrangement bacterial strain HDFS4-26 and breeding method of high-yield bacterial strain

A technology of genome rearrangement, high-yield strains, applied in the directions of microorganism-based methods, biochemical equipment and methods, fungi, etc., can solve the problems of low yield of paclitaxel, and achieve the effect of simple breeding and good effect

Inactive Publication Date: 2008-10-08
HEILONGJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to solve the problem that the paclitaxel yield of the strains currently available for paclitaxel microbial fermentation is still low, and provide the paclitaxel genome rearrangement strain HDFS4-26 and the paclitaxel high-yielding strain breeding method

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  • Paclitaxel genome rearrangement bacterial strain HDFS4-26 and breeding method of high-yield bacterial strain
  • Paclitaxel genome rearrangement bacterial strain HDFS4-26 and breeding method of high-yield bacterial strain
  • Paclitaxel genome rearrangement bacterial strain HDFS4-26 and breeding method of high-yield bacterial strain

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specific Embodiment approach 1

[0020] Specific embodiment 1: In this embodiment, the paclitaxel genome rearrangement bacterial strain HDFS4-26 is Nodulisporium sylviforme (Nodulisporium sylviforme), which is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation date is On February 28, 2008, the preservation number is CCTCCM 208026; the colony of paclitaxel genome rearrangement strain HDFS4-26 is white at the initial stage of culture, and later it is dark grayish brown, and the back is black; the paclitaxel genome rearrangement strain HDFS4-26 mycelium is partly endogenous , partly superficial, septated, multi-branched, mycelium is dark brown and gradually becomes shallower toward the apex, with a diameter of 2.24-5.86 μm, no sclerotium is seen; conidiophores are solitary, most of which are tree-like branches, Brown to the top, 37-162 μm in length; sporulation cells columnar, solitary or wheel-shaped, 6.37-22.63 μm×2.51-3.86 μm, zygospinous; conidia solitary, oval or egg Rou...

specific Embodiment approach 2

[0025] Specific embodiment 2: The difference between this embodiment and specific embodiment 1 is: paclitaxel genome rearrangement bacterial strain HDFS4-26 was cultured on PDA medium at 25-28°C for 3 days, and the colony was flat and the diameter of the colony was 4.1-5.8 cm . Others are the same as the first embodiment.

specific Embodiment approach 3

[0026] Specific embodiment three: the present embodiment paclitaxel high-yielding bacterial strain is selected and bred according to the following methods: 1, protoplast preparation; 2, the inactivation of protoplast; 3, the fusion of protoplast; 4, protoplast regeneration; Screening to obtain a high-yield paclitaxel strain; the strain used for protoplast preparation is paclitaxel-producing strain HD 100-23 , UV 40-19 and UL 50-6 .

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Abstract

The invention relates to paclitaxel genome shuffling bacterial strain HDFS4-26 and a seed selection method of high yield paclitaxel bacterial strain, which belongs to the paclitaxel genome shuffling bacterial strain and the seed selection method of the paclitaxel engineering bacterial strain and solves the problem that the paclitaxel output of the bacterial strain which can be used in the paclitaxel microorganism fermentation at present is still relatively low. The paclitaxel genome shuffling bacterial strain HDFS4-26 is nodulisporium sylviforme and belongs to the nodulisporium and is preserved in the China Center for Type Culture Collection, the preservation date is February 28, 2008, the preservation number is CCTCC M 208026. The seed selection method of the high yield paclitaxel bacterial strain adopts firstly, the protoplast preparation; secondly, the inactivation of the protoplast; thirdly, the interfusion of the protoplast; fourthly, the regeneration of the protoplast; fifthly, the genome shuffling and sieving, therefore to obtain the high yield paclitaxel bacterial strain. The paclitaxel output of the bacterial strain HDFS4-26 is 516.37ug / L. The seed selection method of the high yield paclitaxel bacterial strain provided by the invention is simple, the effect is good, and the paclitaxel genome shuffling bacterial strain HDFS4-26 and the seed selection method of high yield paclitaxel bacterial strain lay the foundation of industrial production of bacterial strain providing high yield paclitaxel.

Description

technical field [0001] The invention relates to a paclitaxel genome rearrangement strain and a method for breeding paclitaxel engineering strains. Background technique [0002] Malignant tumors seriously threaten human health, and cancer is the second leading cause of death in human diseases after cardiovascular and cerebrovascular diseases. [0003] Paclitaxel is a diterpene alkaloid drug (molecular formula such as figure 1 shown). Because yew (also known as yew) is a rare tree species, its growth rate is slow, and its biological characteristics have high requirements on the ecological environment, and its natural renewal ability is poor, so the population is extremely limited. It is an endangered plant and its resources are very limited. Therefore, Scholars at home and abroad are actively researching and developing new drug sources of paclitaxel. [0004] Microbial fermentation production is the most ideal way to reduce costs and obtain paclitaxel in large quantities. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/15C12N15/04C12P17/02C12R1/645
Inventor 周东坡赵凯平文祥张丽娜刘军林燕
Owner HEILONGJIANG UNIV
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