Recombinant strain as well as construction method and application in producing campesterol thereof

A kind of recombinant strain, campesterol technology, applied in the field of genetic engineering

Inactive Publication Date: 2017-08-22
TIANJIN UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0005] Therefore, how to adjust the balance between the two metabolic fluxes is the key point to increase the accumulation of terpenes in Yarrowia lipolytica, but there is no report on the regulation and balance of the metabolic flux of Yarrowia lipolytica

Method used

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  • Recombinant strain as well as construction method and application in producing campesterol thereof
  • Recombinant strain as well as construction method and application in producing campesterol thereof
  • Recombinant strain as well as construction method and application in producing campesterol thereof

Examples

Experimental program
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Embodiment

[0057] Example High-yield campesterol Recombinant Yarrowia lipolytica strain Acetyl-CoA metabolism regulation and optimization

[0058] The construction and characterization method of the recombinant Yarrowia lipolytica strain of high-yield campesterol of the present invention are as follows:

[0059] 1. Acquisition of recombinant Yarrowia lipolytica for the synthesis of campesterol

[0060] The ergosterol synthesized in the wild-type Yarrowia lipolytica strain cannot be transformed into downstream steroid hormones. The synthesis of campesterol in yeast cells needs to cut off the endogenous ergosterol production pathway and knock out sterol C-22 Reductase (ERG5), and the introduction of the C7 reductase (DHCR7) gene at the same time can shift the metabolic flow from producing ergosterol to campesterol. It is consistent with the construction method of Yarrowia lipolytica that has reported the highest yield of campesterol (such as figure 1 ), select the DHCR7 gene from zebrafi...

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Abstract

The invention relates to the technical field of genetic engineering, in particular to a recombinant strain as well as a construction method and application in producing campesterol thereof. The invention constructs a recombinant strain expressing a DHCR7 gene and an ERG5 gene with inactivation. The strain also over-expresses an acetyl-CoA metabolism-related gene, the recombinant yarrowia lipolytica chassis strain increases the yield of the campesterol by regulating the metabolic flow of the acetyl-CoA synthesis and fatty acid synthesis in the cytoplasm, and the experiments show that the yield of campesterol of the recombinant strain can reach 941.5mg / L after process optimization.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a recombinant bacterial strain and its construction method and its application in producing campesterol. Background technique [0002] Steroid hormone drugs are the second class of drugs next to antibiotics, and are widely used in the treatment of respiratory diseases, endocrine disorders, lymphatic leukemia, and rheumatism. As an important precursor of steroid hormone drugs, campesterol has important applications in the synthesis of many steroid hormone drugs such as glucocorticoids and sex hormones. The traditional preparation of campesterol mostly adopts the method of plant extraction. This method has cumbersome operation process, low yield and serious organic pollution, which greatly limits the further production and application of campesterol. [0003] The use of microbial synthesis is considered to be a promising production method with low cost, high yield, low...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/81C12P33/00C12R1/645
Inventor 王颖张羽姚明东肖文海刘宏周晓元英进
Owner TIANJIN UNIV
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