A kind of pseudonocardia and its application

A pseudonocardia and cell technology, applied in bacteria, microorganisms, microorganisms, etc., can solve the problems of long transformation cycle, single transformation substrate, low strain transformation efficiency, etc., to improve transformation efficiency, reduce difficulty and cost. Effect

Active Publication Date: 2021-07-27
NANJING HERON PHARMA SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the current reports, Pseudonocardia and Streptomyces catalyze the transformation of calcitriol and calcifediol, and all of them have the problems of low strain transformation efficiency, long transformation cycle, and single transformation substrate

Method used

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  • A kind of pseudonocardia and its application
  • A kind of pseudonocardia and its application
  • A kind of pseudonocardia and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] The pseudonocardia HRW001 glycerol tube strain was inserted into the solid plate medium for activation, the culture temperature was 25°C, and the culture was static for 2 days. The activated solid plate strains were placed in a 1L shake flask for primary seed expansion cultivation, with a liquid volume of 300mL, a cultivation temperature of 25°C, a rotational speed of 100rpm, and a cultivation period of 2 days. Put the primary seed liquid into a 50L secondary seed tank according to the inoculation amount of 1% for secondary seed expansion cultivation, the liquid volume is 30L, the cultivation temperature is 25°C, the rotation speed is 100rpm, the ventilation rate is 0.5vvm, and the tank pressure is 0.02MPa , the culture period is 2 days; according to the inoculum amount of 5%, put the secondary seed liquid into a 200L fermenter for fermentation culture, the liquid volume is 150L, ​​the culture temperature is 25°C, the initial pH is 6.5, and 20% is added after fermentatio...

Embodiment 2

[0058] The pseudonocardia HRW001 glycerol tube strain was inserted into the solid plate medium for activation, the culture temperature was 28°C, and the culture was static for 4 days. The activated solid plate strains were placed in a 1L shake flask for primary seed expansion cultivation, with a liquid volume of 300mL, a cultivation temperature of 28°C, a rotational speed of 150rpm, and a cultivation period of 3 days. Put the primary seed solution into a 50L secondary seed tank according to the inoculation amount of 2% to carry out secondary seed expansion cultivation. The liquid volume is 30L, the cultivation temperature is 28°C, the rotation speed is 300rpm, the ventilation rate is 1vvm, and the tank pressure is 0.03MPa. The culture period is 3 days; according to the inoculum amount of 10%, put the secondary seed liquid into a 200L fermenter for fermentation and culture, the liquid volume is 150L, ​​the culture temperature is 28°C, the initial pH is 6.5, and 20% ammonia water...

Embodiment 3

[0065] The pseudonocardia HRW001 glycerol tube strain was inserted into the solid plate medium for activation, the culture temperature was 33°C, and the culture was static for 3 days. The activated solid plate strains were placed in a 1L shake flask for primary seed expansion cultivation, with a liquid volume of 300mL, a cultivation temperature of 33°C, a rotational speed of 150rpm, and a cultivation period of 3 days. Put the primary seed liquid into a 50L secondary seed tank according to the inoculum amount of 3% for secondary seed expansion cultivation, the liquid volume is 30L, the cultivation temperature is 33°C, the rotation speed is 200rpm, the ventilation rate is 1.5vvm, and the tank pressure is 0.03MPa , the culture period is 2 days; according to the inoculum amount of 15%, the secondary seed liquid is put into a 200L fermenter for fermentation and culture, the liquid volume is 150L, ​​the culture temperature is 33°C, the initial pH is 6.5, and 20% is added after fermen...

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Abstract

The present invention relates to the technical field of microbes, in particular to the application of Pseudonocardia in microbial conversion to catalyze the hydroxylation of sterol compounds. The original strain of Pseudonocardia is obtained by separating and purifying the soil of Nanjing Laoshan National Forest Park , and screened to obtain Pseudonocardia HRW001 of the present invention after compound mutagenesis by ultraviolet light, microwave, diethyl sulfate and protoplast fusion technology. It was identified by the China General Microorganism Culture Collection Management Center and classified as Pseudonocardia Pseudonocardia sp.HRW001, the deposit number is CGMCC No.17524. The invention obtains a large amount of bacterium after liquid fermentation of pseudonocardia, carries out microbial transformation and catalyzes the hydroxylation of sterol compounds, and has high catalytic activity and specificity. The pseudonocardia of the present invention is an important microorganism for biotransforming and catalyzing the hydroxylation of sterol compounds, can catalyze the hydroxylation of various sterol compounds, and has great application value.

Description

technical field [0001] The invention belongs to the technical field of microbial transformation, and in particular relates to Pseudonocardia sp. HRW001 and its application for microbial transformation to catalyze the hydroxylation of sterol compounds. Background technique [0002] Pseudonocardiaceae are a group of evolutionarily related, morphologically diverse, peptidoglycan containing Meso-DAP (meso-diaminopimelic acid), galactose and arabinose in the whole cell hydrolyzate, no branch Bacteric acid actinomycetes. Many members of this family can ferment to produce important biologically active substances, including antibiotics, enzymes, vitamins, etc. At the same time, there are very few pathogenic bacteria in this family, which is a kind of safe microorganism. [0003] Cytochrome P450 enzymes (P450s or CYPs) are a class of multifunctional enzymes that depend on heme and catalyze a variety of complex oxidation reactions. It can catalyze a variety of substrates and widely...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P33/06C12R1/01
CPCC12N1/20C12P33/06C12N1/205C12R2001/01
Inventor 文昌王辉焦雪茜郑慧娟
Owner NANJING HERON PHARMA SCI & TECH CO LTD
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