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Epidermis brevibacterium ZJB-07021 and use for preparation of (S)-2,2-dimethyl cyclopropane formamide thereof

A technology of dimethylcyclopropanecarboxamide, ZJB-07021, which is applied to Brevibacterium epidermidis ZJB-07021 and its application in the preparation of (S)-2,2-dimethylcyclopropanecarboxamide, which can solve the problem of Environmental hazards and other issues

Active Publication Date: 2008-05-21
ZHEJIANG UNIV OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Using traditional chemical methods to prepare optically pure (S)-2,2-dimethylcyclopropanecarboxamide requires adding a large amount of solvents and expensive and toxic chiral catalysts in the reaction, which will cause great harm to the environment

Method used

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  • Epidermis brevibacterium ZJB-07021 and use for preparation of (S)-2,2-dimethyl cyclopropane formamide thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Isolation of strains

[0037] Isolation of R-amidase-producing bacteria: Add 1 g of soil sample and appropriate amount of glass beads to 9 mL of 0.85% saline, shake well to make a uniform soil suspension; pipette 0.5 mL of soil suspension to inoculate In a 250-mL Erlenmeyer flask with 30 mL of enriched medium, place it in a shaker at 30°C and 150 rpm for 4 days. After the enriched liquid appears turbid, transfer 0.5 mL to fresh medium and continue to culture for 4 days ; After repeating 4 to 5 cycles, the enrichment solution is diluted with multiple gradients and spread on the separation plate to obtain a single colony;

[0038] The enrichment medium uses 2,2-dimethylcyclopropanamide as the only nitrogen source, and the composition is as follows (final concentration): K 2 HPO 4 ·12H 2 O: 2.5g / L, KH 2 PO 4 :2g / L, MgSO 4 ·7H 2 O: 0.5g / L, FeSO 4 ·7H 2 O: 0.01g / L, CaCl 2 :0.06g / L, 2,2-Dimethylcyclopropanecarboxamide: 1.5g / L, prepared with tap water, pH adjusted to 7....

Embodiment 2

[0040] Example 2: Cultivation of strains

[0041] Slant medium composition (final concentration): beef extract 5g / L, peptone 10g / L, NaCl5g / L, agar 18g / L, prepared with tap water;

[0042] Pick a single colony of CCTCC M.207076 on the plate, inoculate it on the slope, cultivate it in a constant temperature incubator at 30°C for 48 hours, and store it in a refrigerator at 4°C for later use.

Embodiment 3

[0043] Example 3: Obtaining wet bacterial cells

[0044] Medium preparation: glucose 10.0g, yeast powder 13.0g, KH 2 PO 4 0.5g, K 2 HPO 4 0.5g, NaCl1g, acetamide 5g, tap water to make up to 1L, pH6.5;

[0045] 250ml shake flask with 30% liquid volume, inoculate a loop of Brevibacterium epidermidis CCTCC M.207076, shake culture at 20°C, 300rpm for 72h; after the culture, the fermentation broth is centrifuged and washed twice with saline to collect the wet bacterial cells and suspended in 50mM In the phosphate buffer solution of pH 7.8, a bacterial suspension with a wet bacterial cell content of 0.5 g / mL is obtained for use.

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Abstract

The invention provides a novel strain which is Brevibacterium epidermidis ZJB-07021 and the application in the microbiological preparation of (S)-2, 2-dimethylcycloproprane carboxamide. The beneficial effects of the invention mainly lie in that the invention provides a novel strain which has stereo selectivity in the production of optically pure (S)-2, 2-dimethylcycloproprane carboxamide; and the strain can be used for the production of optically pure (S)-2, 2-dimethylcycloproprane carboxamide. The invention obtains a novel strain which is different from the previous research through screening; the invention provides the basis for the investigation on the variety of microbial strain in the chiral resolution aspect and the further comparison of the difference of transformation pathway and reaction mechanism among various strains.

Description

(1) Technical field [0001] The invention relates to a new strain-Brevibacterium epidermidis ZJB-07021 (Brevibacterium epidermidis ZJB-07021), and its application in the preparation of (S)-2,2-dimethylcyclopropane carboxamide by microorganisms. (2) Background technology [0002] The chemical structure of 2,2-dimethylcyclopropane carboxamide is: [0003] [0004] The racemic 2,2-dimethylcyclopropanecarboxamide consists of two enantiomers of (S)-2,2-dimethylcyclopropanecarboxamide and (R)-2,2-dimethylcyclopropanecarboxamide Body composition. [0005] (S)-2,2-Dimethylcyclopropanecarboxamide is a key chiral intermediate in the synthesis of cilastatin. Cilastatin, the chemical name is (Z)-7-[(2R)-(2amino-2-carboxyethyl)sulfur]-[(1S)-2,2-dimethylcyclopropanecarboxamido] -2-Heptenoic acid, is the first clinical application of renal dehydropeptidase inhibitor. Tienam, a compound preparation of cilastatin and imipenem, is a broad-spectrum β-lactam antibiotic developed by Merck in 1979. I...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P13/02C12R1/13
Inventor 郑裕国郑仁朝金少军王远山王宝峰沈寅初
Owner ZHEJIANG UNIV OF TECH
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