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Bacterial strain, construction method and application thereof

A strain and campesterol technology, applied in the field of microorganisms, can solve the problems of inability to meet the needs of industrial production and low content of campesterol, and achieve the effects of low cost, less by-products and less pollution

Active Publication Date: 2016-12-07
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the content of campesterol in shake flask fermentation of recombinant strains obtained at present is low, which cannot meet the needs of industrial production.

Method used

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  • Bacterial strain, construction method and application thereof
  • Bacterial strain, construction method and application thereof
  • Bacterial strain, construction method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0042] Source Screening of Exogenous Functional Gene dhcr7 Synthesized by Recombinant Saccharomyces cerevisiae

[0043] The construction method of the recombinant Saccharomyces cerevisiae strain producing campesterol of the present invention is as follows:

[0044] 1. Acquisition of exogenous functional gene elements

[0045] The exogenous gene is the key enzyme C7 reductase DHCR7 gene used for the synthesis of campesterol: select 11 functional genes from different sources to screen the optimal gene source for campesterol synthesis. The gene sources of DHCR7 include Rattus norvegicus, small Mouse (Mus musculus), human (Homo sapiens), common marmoset (Callithrix jacchus), cow (Bos taurus), jungle chicken (Gallus gallus), African clawed frog (Xenopus laevis), flat algae (Tetraselmissp.GSL018), Chlamydia ( Waddlia chondrophila WSU 86-1044)、(CandidatusProtochlamydia amoebophila UWE25)、斑马鱼(Danio rerio)等,依次简写为Sc_Rn_DHCR7、Sc_Mm_DHCR7、Sc_Hs_DHCR7、Sc_Cj_DHCR7、Sc_Bt_DHCR7、Sc_Gg_DHCR7、S...

Embodiment 2

[0079] Source Screening of Exogenous Functional Gene dhcr7 Synthesized by Recombinant Yarrowia lipolytica

[0080] The construction method of the recombinant Yarrowia lipolytica strain producing campesterol of the present invention is as follows:

[0081]1. Acquisition of exogenous functional gene elements

[0082] The exogenous gene is the key enzyme C7 reductase DHCR7 gene for the synthesis of campesterol: select 6 functional genes from different sources to screen the optimal gene source for campesterol synthesis, including campesterol screened by Yuan Yingjin's research group at Tianjin University Xenopus laevis (Xenopus laevis) DHCR7 gene with the highest yield. Gene sources of DHCR7 include Xenopus laevis (Xenopus laevis), human (Homo sapiens), jungle fowl (Gallus gallus), zebrafish (Danio rerio), Arabidopsis ( Arabidopsis thaliana), Chlamydia (Waddlia chondrophila), etc., are abbreviated as Yl_Xl_DHCR7, Yl_Hs_DHCR7, Yl_Gg_DHCR7, Yl_Dr_DHCR7, Yl_At_DHCR7, Yl_Wc_DHCR7, an...

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Abstract

The invention relates to the field of microorganisms and especially relates to a bacterial strain, a construction method and an application thereof. The invention provides the bacterial strain for high-yield campesterol. An exogenous functional gene DHCR7 is guided into a chassis bacterial strain. Compared with plant extraction, the method for utilizing two yeast chassis bacterial strains including recombined brewer yeast and lipid yeast solution to produce campesterol has the advantages of low cost, few side products and low pollution; a feasible method is supplied for the industrial production of the campesterol. The basis is established for the biosynthesis of steride hormone drugs including downstream progesterone, 4AD and the like. The bacterial strain has guidance significance in acquiring high-yield campesterol through other microorganisms.

Description

technical field [0001] The present invention relates to the field of microorganisms, in particular to bacterial strains and their construction methods and applications. Background technique [0002] At present, steroid hormone drugs are the second class of drugs next to antibiotics, and are widely used in the preparation of health products, treatment of respiratory diseases, endocrine disorders, lymphatic leukemia, rheumatism and skin diseases. As an important intermediate of steroid hormone drugs, campesterol can be used as a precursor for the synthesis of important steroid intermediates such as 4AD, ADD, 9α-OH-AD, and important steroid drugs such as progesterone and hydrocortisone. It has important applications in the field of pharmaceutical synthesis. Campesterol is mostly prepared by plant extraction, but the plant extraction process has many steps, low yield, complicated separation of by-products, and easy pollution to the environment, which greatly limits the producti...

Claims

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Application Information

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IPC IPC(8): C12N1/19C12P33/00C12R1/865C12R1/645
CPCC12N9/001C12P33/00C12Y103/01021
Inventor 李霞张羽肖文海王颖刘宏元英进
Owner TIANJIN UNIV