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Coxakia rhizotrophis ZJPH202011 and application thereof

A technology of ZJPH202011, Saxobacteria, which is applied in the direction of bacteria, microorganisms, and methods based on microorganisms, etc., can solve the problems of reduced activity of cysteine ​​proteases and the number of worms in the body, and achieve high concentration of catalytic substrates and transformation process Green, environment-friendly effect

Active Publication Date: 2021-10-01
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the development of the drug was stopped because it was found to increase the risk of stroke in patients in clinical studies, the search for new uses of the drug is still ongoing. Jon J. Vermeire and others discovered Odanacatib in the process of finding a drug for hookworm disease It can reduce the number of worms in the body by 73%, and at the same time reduce the activity of cysteine ​​protease in the parasite by 51%, which provides a new possibility for the development of low-dose drugs and alternatives for the treatment of hookworm disease

Method used

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  • Coxakia rhizotrophis ZJPH202011 and application thereof
  • Coxakia rhizotrophis ZJPH202011 and application thereof
  • Coxakia rhizotrophis ZJPH202011 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: strain screening and identification

[0027] 1. The source of the strain: Kosakonia radicincitans ZJPH202011 was isolated and screened from soil samples of cornfields in Yanwu Town, Fenyang City, Shanxi Province. The specific screening methods are as follows:

[0028] Primary screening: Weigh 2g of soil sample into a 250mL Erlenmeyer flask filled with 100mL of normal saline with a mass concentration of 0.9%, vibrate in a shaker at 30°C and 200rpm for 30min, then take 2mL of soil sample suspension and add it to 100mL of seed flask culture medium in a 250mL conical flask at 30°C and 200rpm for 24h. After the culture medium became turbid, transfer 2mL of the bacterial liquid to a 250mL Erlenmeyer flask containing 100mL enrichment medium and cultivate for 3-5d. The enrichment medium uses 4'-bromo-2,2,2-trifluoroacetophenone as the sole carbon source, and its final concentration is composed of: 4'-bromo-2,2,2-trifluoroacetophenone 10 mM , (NH 4 ) 2 SO 4 2.0...

Embodiment 2

[0048] Embodiment 2: the acquisition of wet thalline

[0049] Slant medium composition: glucose 15g / L, peptone 20g / L, yeast extract 10g / L, (NH 4 ) 2 SO 4 2g / L, KH 2 PO 4 2g / L, NaCl 1g / L, MgSO 4 0.244g / L, agar 20g / L, solvent is water, pH 6.5;

[0050] Seed medium composition: glucose 15g / L, peptone 20g / L, yeast extract 10g / L, (NH 4 ) 2 SO 4 2g / L, KH 2 PO 4 2g / L, NaCl 1g / L, MgSO 4 ·7H 2 O 0.5g / L, solvent is water, pH 6.5;

[0051] Fermentation medium composition: sucrose 25g / L, (NH 4 ) 2 SO 4 5g / L, KH 2 PO 4 2g / L, the solvent is water, pH 6.5.

[0052] 1) Slant culture: Pick a single colony of Cossackia root-promoting Cossackia ZJPH202011 and inoculate it into the slant medium, cultivate it at 30°C for 24 hours, pick a single colony on the slant medium and inoculate it for 24 hours, and repeat twice to obtain the slant Store in a refrigerator at 4°C;

[0053] 2) Seed culture: Pick a ring of bacteria from the slant and put it into 100mL seed medium, cul...

Embodiment 3

[0056] The wet thalline obtained according to the method of Example 2 is resuspended in 10mL of potassium phosphate buffer (0.1M, pH 7.0), and the amount of wet thalline is 100g / L buffer in terms of wet weight; 20mM 4'-bromo-2,2,2-trifluoroacetophenone as the substrate, then add 100g / L glucose as the auxiliary substrate based on the buffer volume, and place it in a shaker at 30°C and 200rpm Reaction 24h. The detection method of Example 1 was used for analysis and detection, and the measurement results were as follows: the concentration of the product (R)-1-(4-bromophenyl)-2,2,2-trifluoroethanol was 9.23 mM, and the concentration of (S)-1- The concentration of (4-bromophenyl)-2,2,2-trifluoroethanol is 0.37mM, and the residual concentration of the substrate 4'-bromo-2,2,2-trifluoroacetophenone is 9.26mM. The above detection results It can be calculated that the e.e. value of the product (R)-1-(4-bromophenyl)-2,2,2-trifluoroethanol is 92.26%, and the yield is 46.14%. The substra...

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Abstract

The invention relates to coxakella rhizotrophy ZJPH202011 and application thereof, and provides a new strain which can be used for catalyzing 4 '-bromo-2, 2, 2-trifluoroacetophenone to prepare (R)-1-(4-bromophenyl)-2, 2, 2-trifluoroethanol through asymmetric reduction, and the (R)-1-(4-bromophenyl)-2, 2, 2-trifluoroethanol prepared through catalysis of the strain has the advantages of being high in catalytic substrate concentration, high in product optical purity and the like. Compared with the existing reported level, the enantiomer excess value of the product obtained by catalysis of the wild strain is high, the e.e. value is greater than 90.66%, the microbial strain is easy to culture and rich in source, expensive coenzyme does not need to be added in the conversion process, and the method has the characteristics of low cost, green conversion process, environmental friendliness and the like. According to the present invention, the 4 '-bromo-2, 2, 2-trifluoroacetophenone is subjected to reduction by using the Kosakonia radicincts ZJPH202011 cell as the catalyst, and when the substrate concentration is 30 mM, the e.e. value of the target product (R)-1-(4-bromophenyl)-2, 2, 2-trifluoroethanol is more than 93.82%, and the yield reaches 75.54%;.

Description

(1) Technical field [0001] The present invention relates to a new microbial strain—Kosakonia radicincitans (Kosakonia radicincitans) ZJPH202011, and its preparation by catalyzing the asymmetric reduction of 4'-bromo-2,2,2-trifluoroacetophenone (R )-1-(4-bromophenyl)-2,2,2-trifluoroethanol. (2) Background technology [0002] The chemical structural formula of (R)-1-(4-bromophenyl)-2,2,2-trifluoroethanol is: [0003] [0004] (R)-1-(4-bromophenyl)-2,2,2-trifluoroethanol is an important intermediate in the synthesis of tryptophan hydroxylase (TPH) inhibitors and JANUS kinase (JAK) inhibitors. Among them, TPH inhibitors regulate peripheral serotonin levels by blocking the synthesis of 5-hydroxytryptamine, which can be used to relieve nausea and vomiting caused by chemotherapy and treat irritable bowel syndrome, and its representative drug is LX1031. As an oral drug, LX1031 is a locally acting TPH inhibitor, which can improve symptoms such as abdominal pain, cramps, constipa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/22C12R1/01
CPCC12N1/20C12P7/22
Inventor 王普孔祥鑫桂铅于双嵘
Owner ZHEJIANG UNIV OF TECH
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