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Method for producing R-mandelic acid and derivates thereof by biocatalysis

A biocatalysis method, mandelic acid technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of difficult industrialization, small processing capacity, low activity, etc., and achieve easy industrial production and mild reaction conditions. , the effect of high hydrolase activity

Active Publication Date: 2012-05-30
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

①Diastereomeric salt crystallization resolution method, the common problem is that it is expensive and has certain toxicity, so it causes waste of resources and environmental pollution to a certain extent
②Chromatographic resolution method, chromatography method has good capacity, selectivity and stability, and the resolution purity is the highest, and plays an increasingly important role in chiral resolution, but the equipment cost of this method is too high, and the consumption is large. The cost is too high and the processing volume is small, so it is limited to detection and laboratory preparation, and cannot be used in commercial production
However, it faces problems such as low activity and difficulty in industrialization

Method used

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  • Method for producing R-mandelic acid and derivates thereof by biocatalysis
  • Method for producing R-mandelic acid and derivates thereof by biocatalysis
  • Method for producing R-mandelic acid and derivates thereof by biocatalysis

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Experimental program
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Effect test

Embodiment 1

[0031] Embodiment 1: Screening and identification of bacterial strains

[0032] 1. Screening of strains

[0033] About 80 soil samples were collected from the sewage outlets of factories, garbage dumps and near the laboratory in various parts of Zhejiang Province for strain screening.

[0034]The composition of the final concentration of the enrichment medium: (NH 4 ) 2 SO 4 1g / L, KH 2 PO 4 0.2g / L, Na 2 HPO 4 0.8g / L, MgSO 4 ·7H 2 O 0.2g / L, CaCl 2 0.1g / L, FeCl 3 0.005g / L, pH7.0, solvent is water.

[0035] The composition of the final concentration of the slant medium: glucose 10g / L, yeast extract 10g / L, K 2 HPO 4 5g / L, MgSO 4 ·7H 2 O 0.2g / L, FeSO 4 0.03g / L, NaCl 1g / L, agar 20g / L, pH7.0, solvent is water.

[0036] The composition of the final concentration of the enzyme production medium: glucose 10g / L, yeast extract 10g / L, K 2 HPO 4 5g / L, MgSO 4 ·7H 2 O 0.2g / L, FeSO 4 0.03g / L, NaCl 1g / L, caprolactam 1g / L, pH 7.0, solvent is water.

[0037] Add 2mM...

Embodiment 2

[0054] 1. Cultivation of cells containing nitrilase

[0055] The composition of the final concentration of the seed medium: ammonium acetate 10g / L, yeast extract 6g / L, dipotassium hydrogen phosphate 5g / L, sodium chloride 1g / L, magnesium sulfate 0.2g / L, and the solvent is water. The CCTCC No: M 208168 strain was inoculated into the seed medium and cultured at 30°C for 28 hours to obtain the seed solution.

[0056] Final concentration composition of enzyme-producing fermentation medium: ammonium acetate 12.14g / L, yeast extract 7.79g / L, n-butyronitrile 3.29g / L, dipotassium hydrogen phosphate 5g / L, sodium chloride 1g / L, magnesium sulfate 0.2g / L, the solvent is water.

[0057] The seed solution was transferred to the optimized fermentation medium with an inoculation amount of 6% (v / v), and the liquid content in a 15L fermenter was 8L. The culture temperature was 30° C., the rotational speed was 150 rpm, and the ventilation volume was 0.4 vvm, and samples were taken to detect bio...

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Abstract

The invention provides a method for producing R-mandelic acid and derivates thereof by biocatalysis. The method comprises performing hydrolysis reaction in a reaction system which takes racemization mandelonitrile compound shown in formula (I) as the zymolyte, and nitrilase obtained by culturing bacillus foecalis alkaligenes (CCTCC No: M 208168) as the catalyst at 20-60 DEG C with pH being 8.0-8.5 to obtain chiral R-mandelic acid and derivates thereof shown in corresponding formula (II). The invention has the beneficial effects that the activity of strain bacillus foecalis alkaligenes is high, the dosage of thallus is little, the produced waste water in the reaction process is little, and pollution is less; the reaction conditions are mild and energy consumption is low; the cost is low, conversion rate is high and yield is high, and the invention is favorable for industrialized production of chiral R-mandelic acid and R-chloro mandelic acid.

Description

(1) Technical field [0001] The invention relates to a method for producing R-mandelic acid and its derivatives by biocatalysis. (2) Background technology [0002] R-mandelic acid is an important pharmaceutical intermediate, widely used in the synthesis of various drugs, such as cephalosporins, penicillins, anti-tumor agents, anti-obesity drugs, optically pure amino acids, angiotensin-converting enzyme inhibitors , Coenzyme A, etc., can also be used to make chiral solvents. Studies have shown that, compared with drugs synthesized by racemic mandelic acid or its derivatives, the drugs synthesized from single-configuration mandelic acid (or derivatives of mandelic acid) not only have higher efficacy, but more importantly, the side effects are reduced Therefore, it must be a single-type compound in many drug synthesis applications; R-mandelic acid is currently the most eye-catching acidic optical resolution agent due to its good biodegradability, which can make most racemic ami...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P41/00C12P7/42C12R1/05
Inventor 郑裕国薛亚平柳志强徐赛珍沈寅初
Owner ZHEJIANG UNIV OF TECH
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