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Construction method and application of plasmid capable of replicating and expressing dehydrogenase gene at positions 1 and 2 of steroid compound a ring in Arthrobacter simplex

A simple Arthrobacter and steroidal compound technology, applied in the field of genetic engineering and microbial fermentation, can solve the problems of undiscovered patent publications, etc., and achieve the effects of low production cost, high conversion rate and mild environment

Inactive Publication Date: 2016-05-25
TIANJIN UNIV OF SCI & TECH
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[0005] Through searching, no patent publications related to this patent application have been found

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  • Construction method and application of plasmid capable of replicating and expressing dehydrogenase gene at positions 1 and 2 of steroid compound a ring in Arthrobacter simplex
  • Construction method and application of plasmid capable of replicating and expressing dehydrogenase gene at positions 1 and 2 of steroid compound a ring in Arthrobacter simplex
  • Construction method and application of plasmid capable of replicating and expressing dehydrogenase gene at positions 1 and 2 of steroid compound a ring in Arthrobacter simplex

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[0032] The present invention will be further described in detail below through the specific examples, the following examples are only descriptive, not restrictive, and cannot limit the protection scope of the present invention with this.

[0033] The plasmid Escherichia coli-Corynebacterium glutamicum shuttle expression vector pXJM19 used in the present invention (Construction and application of new Corynebacterium glutamicum vectors, 1999, Biotechnology techniques 13 (6), 437-441);

[0034] The host bacterium used in the present invention is Arthrobacter Simplex, the preservation number is ACCC12070, which can be used for the biological dehydrogenation of steroid compounds.

[0035] Based on the analysis that the shuttle expression vector pXJM19 can replicate in Arthrobacter simplex, the present invention further verifies that the plasmid can be efficiently expressed in Arthrobacter simplex, and successfully realizes the replication of the pXJM19-KsdD recombinant expression ve...

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Abstract

The invention relates to application of plasmid capable of being copied in arthrobacter simplex and expressing steroid compound A ring 1, 2 dehydrogenase genes to constructing high-efficiency expression steroid compound A ring 1, 2 dehydrogenation arthrobacter simplex engineering bacteria. The plasmid is capable of being copied in arthrobacter simplex and efficiently expressing steroid compound A ring 1, 2 dehydrogenase genes and significantly practical, and basis is provided for further using plasmid pXJM19 to modify arthrobacter simplex through molecular techniques.

Description

technical field [0001] The invention belongs to the field of genetic engineering and microbial fermentation, and relates to a plasmid capable of replicating and highly expressing the 1,2-position dehydrogenase gene (KsdD) of steroid A ring in Arthrobacter simplex. technical background [0002] Steroid drugs have important physiological activities and are widely used clinically. They are the second largest class of drugs after antibiotics. Prednisone acetate can be converted into prednisone acetate by introducing a double bond into the C1 and 2 positions of cortisone acetate. The anti-inflammatory effect can be increased by 3-4 times, and the side effects caused by sodium retention can be reduced. At present, the commonly used methods in the production of steroid drugs include chemical synthesis and microbial transformation. The chemical method for dehydrogenation of the C1 and 2 positions generally uses the selenium dioxide method. However, the microbial conversion method h...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/74C12N15/66C12N1/21C12R1/06
Inventor 路福平田耀张会图刘晓光
Owner TIANJIN UNIV OF SCI & TECH
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