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Method for improving the concentration of substrate in biological resolution L-menthol reaction

A technology of substrate concentration and menthol, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problem of low substrate concentration, achieve the effect of increasing substrate concentration and reducing inhibition

Inactive Publication Date: 2009-09-30
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, our laboratory has acquired Rhizopus chinensis CCTCC M201021, Rhizopus oryzae AS 3.41, Candida parapsilosis CICC 1676, Burkholderia cepacia ATCC 25416, Pseudomonas fluorescens Bacteria strains capable of asymmetrically splitting DL-menthyl acetate were screened out from Pseudomonas fluorescens AS1.823 and Pseudomonas fluorescens AS 1.55, but they had strong product inhibition, making the transformed Substrate concentration is too low, 15g / L dry cell concentration, the maximum substrate concentration for single batch transformation is 1%

Method used

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Experimental program
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Effect test

Embodiment 1

[0026] In 5mL, 50mmol / L Na 2 HPO 4 -KH 2 PO 4 In the buffer (pH7.0), add 0.8g Rhizopus chinensis CCTCC M201021 transformed bacterial cells and 0.25g DL-menthyl acetate, shake the reaction on a constant temperature shaker at 30°C, add 0.25g resin after 2h HZ-816, the whole process took 24 hours. After the reaction, the mixture was centrifuged, and the supernatant was extracted with ethyl acetate to extract the product and substrate. The product was analyzed by chiral gas chromatography. The optical purity of the product L-menthol was 98% ee, and the conversion rate was 46.7%.

Embodiment 2

[0028] In 5mL, 50mmol / L Na 2 HPO 4 -KH 2 PO 4 In the buffer (pH6.0), add 0.8g of Candida parapsilosis CICC 1676 transformed bacterial cells and 0.3g of DL-menthyl acetate, shake the reaction on a constant temperature shaker at 30°C, add 0.3g of resin D- 202, the whole process takes 24 hours. After the reaction, the mixture is centrifuged, and the supernatant is extracted with ethyl acetate to extract the product and substrate, and the product is analyzed by chiral gas chromatography. The optical purity of the product L-menthol is 97.3% ee, and the conversion rate is 46.3%.

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Abstract

The invention relates to a method for improving the concentration of a substrate in a resolution L-menthol reaction, belonging to the technical field of the resolution of a racemic compound by a biological method. In the method, a pH value of an asymmetric resolution DL-acetic acid menthol reaction system is controlled to be from 5.0 to 8.0, conversion strains are obtained in Rhizopus chinensis CCTCC M201021, Rhizopus oryzae AS 3.41, Candida parapsilosis CICC 1676, Burkholderia cepacia ATCC 25416, Pseudomonas fluorescens AS1.823 or Pseudomonas.fluorescens AS 1.55, and any one of resin HZ-801, HZ-816, D-315, D-202, HD-81, NKA-II, D-406 and DA-201 is added to improve the concentration of the single-batch conversion substrate DL-acetic acid menthol up to 6 times.

Description

technical field [0001] The invention discloses a method for increasing the substrate concentration in the reaction of biological resolution of L-menthol, which belongs to the technical field of biological resolution of racemic compounds. Background technique [0002] Menthol is currently one of the most industrially valuable chiral compounds, which are widely used in the fields of medicine, food, cosmetics and chiral reagent preparation. It is very meaningful to carry out the research on the resolution method of menthol. [0003] Natural menthol is affected by weather, region, etc., and its output and quality fluctuate greatly. Synthetic menthol can solve the above problems. In recent years, biological resolution methods have become the mainstream trend of chiral resolution. [0004] At present, our laboratory has acquired Rhizopus chinensis CCTCC M201021, Rhizopus oryzae AS 3.41, Candida parapsilosis CICC 1676, Burkholderia cepacia ATCC 25416, Pseudomonas fluorescens Bac...

Claims

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Application Information

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IPC IPC(8): C12P41/00C12P7/22C12R1/845C12R1/72C12R1/39C12R1/01
Inventor 喻晓蔚徐岩顾鸿
Owner JIANGNAN UNIV
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