Rhodococcus erythropolis XS1012 and application thereof in preparation of chiral alcohol

A technology of XS1012, Rhodococcus erythraea, applied in the direction of bacteria, microorganism-based methods, biochemical equipment and methods, etc., can solve the problem of limited industrial application, complex separation and purification steps of aldehyde-ketone reductase or alcohol dehydrogenase, and cost of coenzymes. expensive, etc.

Active Publication Date: 2014-05-07
ZHONGRONG TECH CORP LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The oxidoreductases used in the asymmetric catalytic reduction of carbonyl compounds are mainly alcohol dehydrogenases or aldehydes and ketones reductases, both of which need to add coenzyme (NAD(P)H) in the catalytic process, but the coenzyme is expensive, and the reduction of aldehydes and ketones The separation and purification steps of enzyme or alcohol dehydrogenase are complicated, which limits its industrial application to a certain extent

Method used

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  • Rhodococcus erythropolis XS1012 and application thereof in preparation of chiral alcohol
  • Rhodococcus erythropolis XS1012 and application thereof in preparation of chiral alcohol
  • Rhodococcus erythropolis XS1012 and application thereof in preparation of chiral alcohol

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Embodiment 1: the acquisition of wet thalline

[0055] Fermentation medium formula: glucose 15g / L, peptone 7.5g / L, yeast extract 6g / L, (NH 4 ) 2 SO 4 3g / L, KH 2 PO 4 1.5g / L, NaCl0.75g / L, MgSO 4 ·7H 2 O0.75g / L, the solvent is water, pH6.5.

[0056] The formula of the seed medium is the same as that of the fermentation medium.

[0057] 1) Slant culture: Pick a single colony of Rhodococcus erythropolis XS1012 and inoculate it into the slant medium, culture at 30°C for 2-3 days, and store in a refrigerator at 4°C; the final concentration of the slant medium is composed of: glucose 15g / L, peptone 7.5g / L , yeast extract 6g / L, (NH 4 ) 2 SO 4 3g / L, KH 2 PO 4 1.5g / L, NaCl0.75g / L, MgSO 4 ·7H 2 O0.75g / L, agar 20g / L, solvent is water, pH6.5.

[0058] 2) Seed culture: Pick a ring of bacteria from the mature slant and insert it into the seed medium, cultivate at 30°C and 200rpm for 24 hours, and obtain the seed liquid;

[0059] 3) Fermentation culture: Transfer the se...

Embodiment 2～4

[0061] The wet thalline of embodiment 1 gained is suspended in the phosphate buffer saline (Na 2 HPO 4 -KH 2 PO 4 buffer), the concentration of the wet bacteria is 35g / L based on the dry weight of the bacteria; add [3,5-bis(trifluoromethyl)phenyl]ethanone with a final concentration of 20mmol / L as a substrate, and then add Glucose with a final concentration of 50g / L was used as an auxiliary substrate, and placed in a shaking table at 30°C and 200rpm for 24 hours. After the reaction, the reaction liquid was centrifuged, and the supernatant was extracted by adding an equal volume of n-hexane. Gas chromatographic analysis, quantitative with internal standard method. The internal standard is dodecane. Take 1ml of the extract and add 2μl of dodecane for analysis. Gas chromatography conditions: Japan Shimadzu GC-2014 gas chromatograph, Zhejiang University N2000 chromatography workstation; American Varian CP-Chirasil-Dex chiral capillary gas chromatography column (25m×0.25mm×0.25...

Embodiment 5～10

[0065] The wet thalline obtained in Example 1 was suspended in 10 mL of different phosphate buffer systems (pH 7.0), and the wet thalline was 35 g / L in terms of dry weight of the thalline; adding [3, 5-bis(trifluoromethyl)phenyl]ethanone is used as substrate, then glucose with a final concentration of 50g / L is added as auxiliary substrate, placed at 30°C, and reacted in a shaker at 200rpm for 24h. After the reaction, the The reaction solution was centrifuged, and the supernatant was added to an equal volume of n-hexane for extraction. The extract was analyzed by gas chromatography and quantified by the internal standard method. The concentration and ee value of the product (S)-[3,5-bis(trifluoromethyl)phenyl]ethanol are shown in Table 2. As can be seen from Table 2, when the buffer system is Na 2 HPO 4 -KH 2 PO 4 , the concentration of (S)-[3,5-bis(trifluoromethyl)phenyl]ethanol was the highest at 14.6mmol / L, the ee value was 99.9%, and the yield was 73.0%.

[0066] The c...

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PUM

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Abstract

The invention discloses a novel microbial strain, namely, rhodococcus erythropolis XS1012 and an application thereof in preparation of (S)-[3,5-bis(trifluoromethyl)phenyl]ethanol. The advantages of high stereoselectivity, high optical purity of a product and the like are realized when the novel strain is used for catalyzing in preparation of optically-pure (S)-[3,5-bis(trifluoromethyl)phenyl]ethanol. Chiral biological catalysis is performed by taking a wet strain obtained by fermenting the rhodococcus erythropolis XS1012 as a catalyst. When the substrate concentration is 20mmol / L, and glucose (50 g / L) and isopropanol (v / v, 10 percent) are taken as auxiliary substrates, the ee of a target product, namely, (S)-[3,5-bis(trifluoromethyl)phenyl]ethanol is up to 99.9 percent, and the yield is up to 92.0 percent.

Description

(1) Technical field [0001] The present invention relates to a new strain of microorganism --- Rhodococcus erythropolis (Rhodococcus erythropolis) XS1012, and preparation of optically pure (S) -[3,5-bis(trifluoromethyl)phenyl]ethanol application. (2) Background technology [0002] The chemical structural formula of (S)-[3,5-bis(trifluoromethyl)phenyl]ethanol is: [0003] [0004] (S)-[3,5-bis(trifluoromethyl)phenyl]ethanol is an important chiral intermediate for the synthesis of new chemotherapy antiemetic drugs NK-1 receptor antagonists, prepared from Antidepressants have good potential for treating a range of central and peripheral nervous system depression. Studies have shown that the activity of [3,5-bis(trifluoromethyl)phenyl]ethanol in S-configuration is significantly higher than that of [3,5-bis(trifluoromethyl)phenyl]ethanol in R-configuration, And it has a strong affinity to NK-1 receptors and has good permeability to the brain barrier. [0005] Currently, the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/22C12R1/01
Inventor 王普李军黄金
Owner ZHONGRONG TECH CORP LTD
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