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Pseudomonas sp. zjut 126 and application in production of L-glufosinate

A zjut126, Pseudomonas technology, applied in microorganism-based methods, bacteria, microorganisms, etc., can solve the problems of high cost and cannot be separated, and achieve the effect of low cost, easy scale and high efficiency

Active Publication Date: 2019-02-12
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These three substances have good water solubility and cannot be separated by simple methods (such as extraction), but can only be separated by ion exchange, which is complex and expensive.

Method used

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  • Pseudomonas sp. zjut 126 and application in production of L-glufosinate
  • Pseudomonas sp. zjut 126 and application in production of L-glufosinate
  • Pseudomonas sp. zjut 126 and application in production of L-glufosinate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The synthesis of embodiment 1 racemic N-decanoyl-(D, L)-glufosinate-ammonium

[0028] Using decanoyl chloride and racemic (D, L)-glufosinate-ammonium to react under alkaline conditions to obtain N-decanoyl-(D, L)-glufosinate-ammonium, the specific experimental steps are as follows:

[0029] (1) Add 0.1mol (21.5g) (D, L)-glufosinate-ammonium ammonium salt to the reactor, adjust pH to 9.0 with mass concentration 20% sodium carbonate aqueous solution, heat to dissolve glufosinate, and cool to 0 in an ice bath ℃, slowly drip 0.12mol decanoyl chloride therein, and the time for dropping is 15 minutes; ~10.0, after the dropwise addition, the temperature was raised to 25°C and then reacted for 2.5h to obtain the original solution;

[0030] (2) Acidify the original solution in step (1) with hydrochloric acid to a pH of 1.0-2.0, and convert N-decanoyl-(D,L)-glufosinate-ammonium into N-decanoyl-(D,L)-glufosinate-ammonium Phosphonic acid, thus separated from water-soluble impurit...

Embodiment 2

[0031] Example 2 Pseudomonas (Pseudomonas sp.) zjut126 strain screening

[0032] 1. Enrichment culture:

[0033] Add the sludge samples collected in the water to 50mL enriched medium with racemic N-decanoyl-(D,L)-glufosinate-ammonium as the sole carbon source and shake culture (30°C, 180rpm) for 1 week, take 5mL The upper turbid liquid was cultured in 50mL fresh enrichment medium for 1 week, and the above operation was repeated, and the concentration of N-decanoyl-(D,L)-glufosinate-ammonium was gradually increased from 0.1g / L in 2 months to 1g / L.

[0034] Enrichment medium: K 2 HPO 4 ·3H 2 O 2.1g / L, KH 2 PO 4 0.4g / L, NaCl 0.1g / L, MgSO 4 ·7H 2 O0.2g / L, CaCl 2 0.025g / L, NH 4 NO 3 0.5g / L, trace element solution 10mL / L, N-decanoyl-(D,L)-glufosinate-ammonium 0.1~1g / L, solvent is deionized water.

[0035] Composition of trace element solution: CoCl 2 0.1g / L, MnSO 4 0.5g / L, FeSO 4 ·7H 2 O 0.1g / L, CuSO 4 0.1g / L, ZnSO 4 ·7H 2 O 0.1g / L, H 3 BO 3 0.01g / L, Al(S...

Embodiment 3

[0042] Example 3 Morphology and Physiological and Biochemical Identification of Bacterial Strain zjut126

[0043] Get the bacterial strain zjut126 that obtains with embodiment 2 and inoculate into solid culture medium (see embodiment 2), microscopic observation, bacterium colony is circular, surface is raised, and edge is neat, moist, milky white, opaque, as time goes on, the growth of bacterium colony There is no obvious change in color form, such as figure 1 shown.

[0044] Perform Gram staining on bacterial strain zjut126, and observe under the microscope, it is a Gram-negative bacterium with a short rod shape, such as figure 2 shown.

[0045] The physiological and biochemical identification of the strain zjut126 is shown in Table 1 and Table 2. The Biolog (GENⅢ) automatic microbial identification system was used for 94 phenotypic tests on the strain, including the detection of 71 carbon source utilization and 23 chemical sensitivities. Detection: Inoculate the strain o...

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Abstract

The invention discloses a pseudomonas sp. zjut126 and application in the production of L-glufosinate. The pseudomonas sp. zjut126 can selectively hydrolyze racemic N-decanoyl-(D, L)-glufosinate to produce L-glufosinate, with selectivity close to 100%. Further, according to the invention, a mixture generated by the reaction is separated by an extraction method, and the raffinate is concentrated andfreeze-dried to obtain an L-glufosinate product. The yield of L-glufosinate extraction separation is 94.6%, the content of L-glufosinate in the product is 92%, and the ee value greater than 95.2%. The method disclosed by the invention has low cost, high efficiency and easiness for scale production.

Description

technical field [0001] The invention relates to a strain of Pseudomonas (Pseudomonas sp.) zjut126 and its application in the field of biocatalysis. The strain has high chiral selectivity for the production of racemic N-decanoyl-(D,L)-glufosinate-ammonium The ability of L-glufosinate-ammonium. With a simple extraction method, high-purity L-glufosinate-ammonium can be isolated from the reaction mixture. Background technique [0002] With the strengthening of environmental protection concepts and the implementation of sustainable development strategies, high efficiency, low toxicity, high activity, and low residue have become an inevitable trend in the development of pesticides. Different enantiomers of some chiral compounds often exhibit different activities. For example, one enantiomer is highly active, while the other enantiomer is inefficient or even has the opposite effect. Glufosinate-ammonium is a high-efficiency, low-toxicity, non-selective (killing) contact herbicid...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P41/00C12P13/04C12R1/38
CPCC12P13/04C12P41/001C12N1/205C12R2001/38
Inventor 张朝晖卫振中史泰龙
Owner ZHEJIANG UNIV OF TECH
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