Arthrobacter simplex engineered strain with excellent stress tolerance and construction method and application thereof

A technology of Arthrobacter simplex and engineering strain, which is applied in the fields of metabolic engineering and microbial transformation, can solve the problems of undiscovered patent documents, etc., and achieves the effects of excellent stress tolerance, improved organic solvent tolerance, and wide distribution of strength

Pending Publication Date: 2020-06-09
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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Problems solved by technology

[0007] Through the search, no published patent documents related

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  • Arthrobacter simplex engineered strain with excellent stress tolerance and construction method and application thereof
  • Arthrobacter simplex engineered strain with excellent stress tolerance and construction method and application thereof
  • Arthrobacter simplex engineered strain with excellent stress tolerance and construction method and application thereof

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Embodiment Construction

[0049] The embodiments of the present invention will be described in detail below. It should be noted that the embodiments are illustrative, not restrictive, and cannot limit the protection scope of the present invention.

[0050] The raw materials used in the present invention, unless otherwise specified, are conventional commercially available products; the methods used in the present invention, unless otherwise specified, are conventional methods in the art.

[0051] A target promoter library applicable to Arthrobacter simplex, said target promoter library is on the basis of expression vector pART2 wild-type promoter hdnOp (SEQ ID NO.2 in the sequence listing), through random mutation and The promoter mutant obtained by site-directed mutagenesis technology, its sequence is: SEQ ID NO.3, and / or is SEQ ID NO.4, and / or is SEQ ID NO.5, and / or is SEQ ID NO.6, and / or for SEQ ID NO.7, and / or for SEQ ID NO.8, and / or for SEQ ID NO.9, and / or for SEQ ID NO.10, and / or for SEQ ID NO.11...

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Abstract

The invention relates to an arthrobacter simplex engineered strain with excellent stress tolerance and a construction method and application thereof. The arthrobacter simplex engineered strain is obtained by adopting a promoter engineering method, a global transcription mechanism engineering method or a method combining the promoter engineering and the global transcription mechanism engineering. The strain still keeps good activity in a transformation system with high concentrations of organic solvents (8%) and substrates (15-45 g/L), with the product yield in a C1,2 (subscript) dehydrogenation reaction being 1.1-2.2 times higher than a control strain I (containing a wild-type promoter and a wild-type irrE). According to the construction method and the application of the engineered strainwith excellent stress tolerance, the concentrations of organic solvents and substrates in the transformation system are increased, and then the product production is increased. The invention has important guiding significance for improving the transformation efficiency of the high-concentration substrates of steroid hydrophobic compounds.

Description

technical field [0001] The invention belongs to the technical field of metabolic engineering and microbial transformation, in particular to a construction method, strain and application of a simple Arthrobacter engineering strain with excellent stress tolerance. Background technique [0002] Steroid hormone drugs are currently the second largest class of drugs in clinical use after antibiotics. They have anti-inflammatory, anti-allergic, anti-shock, and anti-allergic effects. In addition, steroid hormones can also improve protein metabolism, recovery and Enhance physical strength, diuresis and lower blood pressure. The dehydrogenation reaction of C1 and 2 positions of steroidal compounds is a typical representative of industrial production of steroidal drugs by microbial transformation, and it is also the most valuable reaction for the production of prednisolone and its homologues. Arthrobacter simplex is a commonly used steroid C1,2 dehydrogenation reaction strain in domes...

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Application Information

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IPC IPC(8): C12N15/113C12N1/21C12N15/74C12P33/02C12N15/10C12R1/06
CPCC07K14/195C12N15/74C12P33/02C12N15/102
Inventor 骆健美王敏朱文成崔慧林贾红晨申雁冰
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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