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Burkholderia and its application

A technology for Burkholderia grizzi and its application, which is applied in the direction of bacteria, microorganisms, biochemical equipment and methods, and can solve the problems of increasing the cost of catalyst preparation and adding a large amount of cells

Active Publication Date: 2018-03-23
EAST CHINA UNIV OF SCI & TECH +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Patent CN106191193A discloses a method of preparing 5-methoxy-2-((S)-((4-methoxy-3,5-dimethyl-2-pyridyl)methanol by asymmetric oxidation of immobilized Aspergillus niger base) sulfinyl)-1H-benzimidazole, but the cell culture time exceeds 72h, and the amount of cells added is large, and the complicated immobilization process also increases the preparation cost of the catalyst

Method used

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  • Burkholderia and its application
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Experimental program
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Effect test

Embodiment 1

[0049] The screening of embodiment 1 bacterial strain

[0050] (1) Collect 252 soil samples in different environments, including Shanghai Fengxian Chemical Industry Zone, Xinhua Hospital, orchards, near rivers, green belts, campuses, greening of residential areas, Shanghai Botanical Garden, etc. The screening process took four rounds of enrichment culture, and the initial poor medium formula was yeast powder 2g / L, (NH 4 ) 2 SO 4 1.0g / L, K 2 HPO 4 ·3H 2 O 6.0g / L, KH 2 PO 4 3.0g / L, NaCl 0.5g / L, MgSO 4 ·7H 2 O 0.5g / L, CaCl 2 0.05g / L, pH 7.0, the concentration of yeast powder in each round of subculture enrichment culture was halved, and the concentration of substrate (compound OME) (0.1mM in the first round) was doubled in each round of culture, and the screening obtained A target bacterial species capable of utilizing a substrate as a carbon source and transforming it. After culturing for 1-2 days, select a soil sample test tube with turbid culture medium and good ...

Embodiment 2

[0056] Example 2 Identification of Burkholderia glathei (Burkholderia glathei) ECU0712

[0057] (1) Using conventional methods to extract the genomic DNA of the bacterial strain, use the genomic DNA of the bacterial strain as a template, use 16S rDNA amplification general primers to carry out PCR amplification, and detect by agarose gel electrophoresis to amplify the purpose of about 1400bp in size For the fragment, the PCR product was purified and recovered with a gel purification kit (agarose gel DNA recovery kit from Beijing Tiangen Biochemical Co., Ltd.), and finally the recovered DNA fragment was sequenced, and the sequencing result was shown in SEQ ID No.1.

[0058] (2) Comparing the 16S rDNA sequence in the NCBI database, the 16S rDNA similarity between the strain we obtained and Burkholderia glathei (Burkholderia glathei, also known as Caballeronia glathei) is 99%, so we will get The strain was named Burkholderia glathei (Burkholderia glathei) ECU0712.

[0059] (3) Th...

Embodiment 3

[0060] Shake flask culture of embodiment 3 Burkholderia glathei (Burkholderia glathei) ECU0712

[0061] Take the strain slant preserved at 4°C and inoculate it into a test tube LB fermentation medium (peptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, pH 7.2), culture at 30°C and 200rpm with shaking for 10h, and form seed liquid. 2ml seed solution was inoculated into 100ml fermentation medium (5g / L peptone, 5.0g / L yeast extract, 5.0g / L sodium chloride, pH 6.5) at 2% ratio and cultured at 30°C for 24h. Centrifuge and wash to obtain resting cells.

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Abstract

The invention discloses a strain of Burkholderia bacterium, the Burkholderia bacterium is Burkholderia glathei (Burkholderia glathei) ECU0712, and the preservation number is CGMCC NO.14464. Using this strain or its extract as a biocatalyst to catalyze the asymmetric oxidation of thioether to chiral sulfoxide has significant advantages. The obtained product has high optical purity, and has the advantages of simple reaction system, short catalyst preparation time and high product yield. .

Description

technical field [0001] The invention relates to a strain of Burkholderia bacterium and a method for asymmetric oxidation of thioether to synthesize chiral sulfoxide. Background technique [0002] Sodium 5-methoxy-2-((S)-((4-methoxy-3,5-dimethyl-2-pyridyl)methyl)sulfinyl)-1H-benzimidazole, can For the treatment of duodenal ulcers, gastric ulcers, gastritis and gastrointestinal esophagitis. The chemical method uses metal and organic small molecule catalysts to asymmetrically oxidize thioether to synthesize, and has made great progress. However, this method has disadvantages such as excessive oxidation, many by-products, and complicated separation and purification processes. [0003] Use the biological method to asymmetrically oxidize the thioether compound to obtain the chiral sulfoxide of a single enantiomer. This method has less environmental pollution, fewer by-products, good atom economy, and high optical purity of the product. Therefore, using the biological method Asym...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P17/16C12R1/01
CPCC12P17/165C12N1/205C12R2001/01C12N1/20C12P1/04C12P11/00
Inventor 郁惠蕾赵骞张龑潘江许建和
Owner EAST CHINA UNIV OF SCI & TECH
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