Method for whole-cell splitting of racemate beta-lactam by using Bradyrhizobium japonicum

A technology of bradyrhizobium and lactam, applied in the field of separation, can solve the problems of many reaction steps, cumbersome operation, limited enzymes and microorganisms, etc., and achieve the effects of low environmental pollution, high reaction efficiency and high purity

Active Publication Date: 2014-07-23
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the methods reported to obtain chiral pure (-) β-lactams are through organic synthesis, with many reaction steps and relatively cumbersome...

Method used

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  • Method for whole-cell splitting of racemate beta-lactam by using Bradyrhizobium japonicum
  • Method for whole-cell splitting of racemate beta-lactam by using Bradyrhizobium japonicum
  • Method for whole-cell splitting of racemate beta-lactam by using Bradyrhizobium japonicum

Examples

Experimental program
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Effect test

Embodiment 1

[0038] This example is a whole cell of Bradyrhizobium soybean for splitting 3-hydroxy-4-phenyl-2-azetidinone.

[0039] The method comprises the steps of:

[0040] (1) Strain selection: Bradyrhizobium japonicum was selected as the strain, and the strain preservation number was CGMCC No.1.2550; the strain was preserved at -80°C;

[0041] (2) Fermentation broth preparation: inoculate the Bradyrhizobium soybean species into the rhizobia culture medium at a volume ratio of 0.1% to 0.5%, and vibrate at 20°C to 30°C at 220 rpm for 96 to 168 hours;

[0042] Rhizobium liquid fermentation medium: yeast extract 1.0g, mannitol 10.0g, soil extract 200ml, distilled water 800ml, pH7.2;

[0043] Among them, soil extract: 25g soil, add 100ml deionized water, cook at 105kPa for 1 hour, add water after filtering to make up to the volume before cooking;

[0044]The specific preparation method is: dissolve yeast extract, mannitol, and soil extract in deionized water, adjust the pH value to 7.2 w...

Embodiment 2

[0057] This example is a method for producing (-) β-lactam using Bradyrhizobium soybean as the strain.

[0058] The preferred scheme on the basis of the method in example 1, the improvements are:

[0059] (2) In the step, the temperature of the shaking culture is 28°C, the time is 168h, and the rotation speed is 220rpm;

[0060] (3) In the step, the bacterial fermentation liquid after culturing for 168 hours is fully mixed with a phosphate buffer solution with a pH value of 7.0 to obtain a diluted fermentation liquid; the mixing volume ratio of the fermentation liquid and a phosphate buffer solution with a pH value of 7.0 is 1: 1. After fully mixing, centrifuge at 11000rpm for 10min to collect the bacterial sediment; freeze it in a -20°C refrigerator for a certain period of time and then thaw it, then centrifuge at 11000rpm for 10min to collect the wet bacteria and use it as a biocatalyst;

[0061] (4) In the step, the wet bacteria are mixed with 1g / L racemic β-lactam solutio...

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Abstract

The invention relates to a method for whole-cell splitting of a racemate beta-lactam by using Bradyrhizobium japonicum, and belongs to the technical field of separation. According to the present invention, the whole-cell transformation splitting of 3-hydroxy-4-phenyl-2-azetidinone and 3-acetoxy-4-phenyl-2-azetidinone with Bradyrhizobium japonicum has high catalysis efficiency, and the high purity single configuration compound can be obtained, wherein the (3R,4S)3-hydroxy-4-phenyl-2-azetidinone e.e value is 99.99%, the target product relative yield is 99%, the total yield is 49.5%, the (3R,4S)3-acetoxy-4-phenyl-2-azetidinone e e.e value is 99.88%, the target product relative yield is 93.58%, and the total yield is 46.79%.

Description

technical field [0001] The present invention specifically relates to a kind of using soybean bradyrhizobium as a biocatalyst to split racemate 3-hydroxyl-4-phenyl-2-azetidinone and racemate 3-acetoxy-4-benzene The invention discloses a whole cell transformation method of 2-azetidinone, which belongs to the technical field of separation. Background technique [0002] (3R,4S)-3-Hydroxy-4-phenyl-2-azetidinone, referred to as (-)β-lactam, it is a very potential chiral drug intermediate, it has two The isomers are (3R, 4S) configuration and (3S, 4R), and the derivative paclitaxel side chain C-13 of the (3R, 4S) configuration isomer is the most promising new drug for the preparation of cancer clinical chemotherapy One of the important chiral precursors of docetaxel. 3-Acetoxy-4-phenyl-2-azetidinone is the acetylated product corresponding to 3-hydroxy-4-phenyl-2-azetidinone, which can be deacetylated to give 3-Hydroxy-4-phenyl-2-azetidinone. [0003] At present, the main raw ma...

Claims

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Application Information

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IPC IPC(8): C12P41/00C12P17/10C12R1/01
Inventor 郑国钧宋大伟陈清
Owner BEIJING UNIV OF CHEM TECH
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