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Racemization method of N-acetyl-glufosinate

A technology of glufosinate-ammonium and acetyl, which is applied in the field of racemization of N-acetyl-glufosinate-ammonium salt, can solve the problems of low conversion rate, unfavorable environmental protection, high raw material cost, etc., achieve mild and environmentally friendly reaction conditions, save steps, reduce workload effect

Active Publication Date: 2019-07-09
SHANGHAI QIZHOU ZIYUE BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0017] The technical problem to be solved by the present invention is that the existing method for preparing L-glufosinate-ammonium has high raw material cost, low conversion rate, and needs acetic anhydride high-temperature condition reaction to be unfavorable for environmental protection and other defects when adopting chemical isomerization acyl glufosinate-ammonium. The invention provides a racemization method of N-acetyl-glufosinate-ammonium salt

Method used

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  • Racemization method of N-acetyl-glufosinate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1 Acquisition of N-acetylamino acid racemase gene

[0067] The N-acetylamino acid racemase from Amycolatopsis sp.Ts-1-60 and Amycolatopsisorientalis subsp.lurida microorganisms was retrieved from the NCBI database, and the NCBI accession numbers were 4A6G or CAC00653.1, respectively, and two N-acetylamino acids were synthesized from the whole gene racemase gene.

[0068] Table 1 N-acetyl amino acid racemase

[0069] serial number Types of source NCBI accession number Enz.7 AAR-AT Amycolatopsis sp.Ts-1-60 4A6G Enz.8 AAR-AO Amycolatopsis orientalis subsp. lurida CAC00653.1

Embodiment 2

[0070] Example 2 Expression of N-acetylamino acid racemase gene

[0071] The N-acetyl amino acid racemase gene enzyme is linked to pET28a, the restriction site NdeI&HindIII, and the enzyme-linked vector is transformed into the host Escherichia coli BL21 competent cell. The constructed strains were inoculated into TB culture based on 37°C, 200rpm shaker, IPTG concentration 0.1mM for overnight induction, and the bacteria were harvested.

Embodiment 3

[0072] Example 3 The cultivation of N-acetylamino acid racemase thalline and the preparation of crude enzyme solution and the determination of enzyme activity

[0073] Composition of LB liquid medium: peptone 10g / L, yeast powder 5g / L, NaCl 10g / L, dissolved in deionized water and then constant volume, sterilized at 121°C for 20min, ready for use.

[0074] After the engineering bacteria containing the N-acetylamino acid racemase gene were activated by streaking on the plate, a single colony was picked and inoculated into 5ml LB liquid medium containing 50μg / ml kanamycin, and cultured with shaking at 37°C for 12h. Transfer to 150ml fresh LB liquid medium containing 50μg / ml kanamycin according to 2% inoculum amount, shake at 37°C until OD600 reaches about 0.8, add IPTG to its final concentration of 0.5mM, induce culture at 18°C 16h. After the cultivation, the culture solution was centrifuged at 10,000 rpm for 10 min, the supernatant was discarded, and the bacterial cells were col...

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Abstract

The invention discloses a racemization method of N-acetyl-glufosinate. The racemization method includes subjecting the N-acetyl-glufosinate to racemization in the presence of N-acetylamino acid racemase, wherein the N-acetylamino acid racemase is derived from Amycolatopsis sp.Ts-1-60 and / or Amycolatopsis orientalis subsp. Lurida. When the racemization method is applied to preparation of L-glufosinate, the cost can be reduced, operations can be simplified, and reaction conditions can be moderate and environment friendly; the conversion rate can reach more than 50%; the workload for separating and purifying the N-acetyl-glufosinate and glufosinate is reduced; the N-acetylamino acid racemase and deacetylation enzymes can be subjected to in-situ isomerization-resolution by a 'one-pot process',and accordingly, a chemical isomerization step for the N-acetyl-glufosinate and related works for distillation and water removal are omitted.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular, the invention relates to a racemization method of N-acetyl-glufosinate-ammonium salt. Background technique [0002] Glufosinate-ammonium is a broad-spectrum contact herbicide developed by Hearst in the 1980s. At present, the three major herbicides in the world are glyphosate, glufosinate-ammonium, and paraquat. Compared with glyphosate and paraquat, glufosinate-ammonium has excellent herbicidal performance and less side effects. Glufosinate-ammonium has two optical isomers, namely D-glufosinate-ammonium and L-glufosinate-ammonium, but only L-glufosinate-ammonium has herbicidal activity, so the method of developing L-glufosinate-ammonium is important for improving atom economy , It is of great significance to reduce the cost of use and reduce the pressure on the environment. [0003] At present, the methods for preparing L-glufosinate-ammonium mainly include chiral resolution, chemica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P41/00C12P13/00C07F9/30
CPCC12P41/001C12P13/001C07F9/301
Inventor 田振华程占冰丁少南徐艳冰黄瑶
Owner SHANGHAI QIZHOU ZIYUE BIOTECHNOLOGY CO LTD
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