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Enterobacter cloacae ZJPH1903 and application

A ZJPH1903, Enterobacter cloacae technology, applied in bacteria, microorganisms, biochemical equipment and methods, etc., can solve the problems of difficult operation control, high cost limit industrial application, etc., achieve good stereoselectivity, improve catalytic efficiency, product optical high purity effect

Active Publication Date: 2020-04-10
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, transition metals require additional chelating functional groups for high enantioselectivity, difficult operation, and the high cost of transition metals limits their industrial applications.

Method used

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  • Enterobacter cloacae ZJPH1903 and application
  • Enterobacter cloacae ZJPH1903 and application
  • Enterobacter cloacae ZJPH1903 and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1, strain screening, biotransformation and analysis and detection of transformed products, strain identification

[0034] 1) Strain screening:

[0035] The source of the strain: Enterobacter cloacae (Enterobacter cloacae) ZJPH1903 was obtained by isolation and screening from soil samples collected near the pharmaceutical factory in Binjiang District, Hangzhou City, Zhejiang Province. The specific screening method is as follows: add 1g of soil sample to 100ml sterile In a 250ml Erlenmeyer flask of water, shake at 30°C and 200rpm for 0.5h to obtain a soil sample dilution. Take 1ml of the above diluted solution and add it to a 250ml Erlenmeyer flask containing 100ml enriched medium, culture at 30°C and 200rpm for 5 days, after the culture medium becomes turbid, take 1ml of the culture medium and add it to the fresh enriched medium , cultivated under the same conditions for several days, and repeated the enrichment culture for 3 to 4 times. 2,2',4'-Trichloroacetop...

Embodiment 2

[0054] Embodiment 2: the acquisition of wet thalline

[0055] 1) Slant culture: Pick a single colony of Enterobacter cloacae ZJPH1903 and inoculate it into the slant medium, culture at 30°C for 36 hours, and store in a refrigerator at 4°C for later use; the final concentration of the slant medium is composed of: glucose 15g / L, peptone 20g / L 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, agar 20g / L, solvent is water, pH value 6.5.

[0056] 2) Seed culture: Pick a ring of bacteria from the mature slant and insert it into the seed medium, and cultivate it at 30°C and 200rpm for 12 hours to obtain the seed liquid; the formula of the seed medium: glucose 15g / L, peptone 20g / L, yeast Paste 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, the solvent is water, and the pH value is 6.5.

[0057] 3) Fermentation culture: Transfer the seed solution to the fermentation medium with an inoculation ...

Embodiment 3

[0059] The wet bacteria obtained in Example 2 were suspended in 10 mL of different reaction media (pH 7.0), and the concentration of the wet bacteria was 100 g / L reaction medium by wet weight; 2 g / L of 2,2',4'-tri Chloroacetophenone substrate, then add 100g / L glucose as an auxiliary substrate, and place it in a shaker at 30°C and 200rpm for 24h. After the reaction, the reaction solution was centrifuged, and the supernatant was taken, and the detection method of Example 1 was used to measure the yield and optical properties of the product (R)-2-chloro-1-(2,4-dichlorophenyl)ethanol. Purity, as can be seen from Table 1, when the buffer is a phosphate buffer, the yield of (R)-2-chloro-1-(2,4-dichlorophenyl)ethanol is the highest, with a yield of 55.3%, e.e. The value is greater than 99.9%.

[0060] The influence of table 1 different reaction medium on productive rate and e.e. value

[0061]

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Abstract

The invention discloses Enterobacter cloacae ZJPH1903 and application thereof in preparation of (R)-2-chloro-1-(2, 4-dichlorophenyl)ethanol. A method of catalytic preparation of optically pure (R)-2-chloro-1-(2, 4-dichlorophenyl)ethanol with the new strain has the advantages of good stereoselectivity, high product optical purity and the like, and when the substrate concentration is 4g / L, the e.e.Value of the target product (R)-2-chloro-1-(2, 4-dichlorophenyl)ethanol is greater than 99.9%, and the yield is 92.9%. When a surfactant is added into the reaction system, the catalytic efficiency canbe effectively improved, the concentration of the catalytic substrate can be increased to 10g / L, the yield of the target product (R)-2-chloro-1-(2, 4-dichlorophenyl)ethanol reaches 94.3%, and the e.e. Value is greater than 99.9%.

Description

[0001] (1) Technical field [0002] The present invention relates to a new microbial strain—Enterobacter cloacae (Enterobacter cloacae) ZJPH1903, and its preparation of (R)-2-chloro-1-( 2,4-dichlorophenyl) ethanol application. [0003] (2) Background technology [0004] The chemical structural formula of (R)-2-chloro-1-(2,4-dichlorophenyl)ethanol is as follows: [0005] [0006] (R)-2-Chloro-1-(2,4-dichlorophenyl)ethanol is a key chiral intermediate for the synthesis of antifungal drugs. The current clinical antifungal drugs mainly include: miconazole and econazole , tioconazole, fenticonazole and isavuconazole, etc., the optical purity of the chiral hydroxyl block of these antifungal drugs has an important impact on the efficacy of the drug. Imidazoles (miconazole, ketoconazole, etc.) have high antifungal activity, but are limited to external use due to their high toxicity. Fenticonazole nitrate is an imidazole broad-spectrum antifungal drug developed by the original Ita...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P7/22C12R1/01
CPCC12N1/20C12P7/22C12N1/205C12R2001/01
Inventor 王普刘月旺庄文锦钱峰
Owner ZHEJIANG UNIV OF TECH
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