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Aspergillus versicolor ZJB16085 and application of aspergillus versicolor to synthesis of R-2-(4-hydroxyphenoxy) propionic acid

A technology of ZJB16085, hydroxyphenoxy, applied in the field of Aspergillus versicolor by transforming R-2 phenoxypropionic acid into R-2-propionic acid

Active Publication Date: 2019-03-29
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are few foreign research reports on the preparation of R-2-(4-hydroxyphenoxy)propionic acid by biological methods, among which Dingler et al. (Pest Management Science, 2015,46(1):33-35) from BASF have reported Synthesis of R-2-(4-hydroxyphenoxy)propionic acid by hydroxylation of substrate R-2-phenoxypropionic acid by Beauveria bassiana

Method used

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  • Aspergillus versicolor ZJB16085 and application of aspergillus versicolor to synthesis of R-2-(4-hydroxyphenoxy) propionic acid
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  • Aspergillus versicolor ZJB16085 and application of aspergillus versicolor to synthesis of R-2-(4-hydroxyphenoxy) propionic acid

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Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1: Screening of bacterial strain ZJB16085

[0026] (1) Sampling from the soil in many planting areas on the campus of Zhejiang University of Technology, each weighed 1g of soil samples (wet weight), made a suspension with 9mL of 0.9% normal saline, drew 1mL and inoculated it in the In a 250mL Erlenmeyer flask with 50mL enrichment medium containing 10g / L R-2-phenoxypropionic acid, cultivate on a shaker at 28°C and 150rpm for 1-2d for the first enrichment culture; take 1mL of the first The second enrichment culture was carried out in the fresh enrichment medium containing 10g / L R-2-phenoxypropionic acid for the second enrichment culture under the same conditions. The bacterial solution of the second enrichment culture was serially diluted, and 100 μL was taken to dilute 10 3 and 10 5 The diluted solution was spread on the screening medium plate, and cultured in a constant temperature incubator at 28°C for 3-4d. Use a sterile toothpick to pick a single colony...

Embodiment 2

[0036] Embodiment 2: identification of bacterial strain ZJB16085

[0037] (1) Colony morphology observation

[0038] Use an inoculation needle to pick a small amount of spores of the strains preserved on the slant, streak on the PDA plate, and incubate at 28°C for about 7 days. The single colony morphology is shown in image 3 . The diameter of the colony is about 1.8cm, the colony is loose, the hyphae are short, the texture is fluffy, the middle is raised, and the shape is concentric. The color gradually changes from dark green to cyan, and the outermost part is white. The back edge of the colony is white, and the middle gradually turns pale yellow.

[0039] (2) Observation of mycelium morphology

[0040] Spread a piece of circular filter paper on the bottom of a petri dish with a diameter of 9.0 cm, put a U-shaped glass rod on it, put a clean glass slide and two coverslips on the glass rod, cover the dish, and inoculate after sterilization , cultivated at 28°C for about ...

Embodiment 3

[0046] Example 3: Aspergillus versicolor CCTCC No: M 2018408 fermented and transformed R-2-phenoxypropionic acid to produce R-2-(4-hydroxyphenoxy)propionic acid

[0047] Aspergillus versicolor CCTCC NO:M 2018408 was inoculated into the slant medium, and cultured statically at 28°C for 6-7d to obtain slant strains. Pick the bacterium from the slope and inoculate it into a 250mL shake flask filled with 50mL of seed medium, and culture at 28°C and 150-200rpm for 2-3d to obtain a seed liquid. The seed solution was inoculated into a 250mL shake flask containing 50mL of fermentation transformation medium with an inoculum volume concentration of 3%, and cultivated at 28°C and 150-200rpm for 2-3d until the wet cell content in the fermentation broth reached 20g / L, and added the final R-2 phenoxypropionic acid at a concentration of 10g / L, continued to cultivate at 28°C and 150-200rpm for 6-7d. The reaction solution was centrifuged at 10000rpm for 10min, and the supernatant was filtered...

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Abstract

The invention discloses aspergillus versicolor ZJB16085 and an application of the aspergillus versicolor to the synthesis of R-2-(4-hydroxyphenoxy) propionic acid. The preservation number of the aspergillus versicolor is CCTCC NO: M 2018408. The present invention separates out the aspergillus versicolor CCTCC NO: M 2018408 from humic soil, and the aspergillus versicolor can efficiently convert R-2-phenoxypropionic acid and synthesize R-2-(4-hydroxyphenoxy) propionic acid, and can synthesize 4.5 g / L R-2-(4-hydroxyphenoxy) propionic acid in the presence of 10 g / L substrate, wherein the conversion rate is 45%.

Description

technical field [0001] The invention relates to a microorganism which converts R-2 phenoxy propionic acid to synthesize R-2-(4-hydroxyphenoxy) propionic acid and a screening method thereof, belongs to the technical field of bioengineering, and particularly relates to a strain which transforms R-2 Phenoxypropionic acid synthesis of R-2-(4-hydroxyphenoxy)propionic acid by Aspergillus versicolor. Background technique [0002] R-2-(4-hydroxyphenoxy)propionic acid is a key intermediate in the synthesis of phenoxycarboxylic acids and aryloxyphenoxypropionic acid herbicides, and can be used for the synthesis of oxaprop-methyl (soybean, rice ), Fluazifop (rape, potato), clodinafop-propargyl (cereal) and other herbicides. As phenoxycarboxylic acid herbicides are getting more and more attention, its advantages as herbicides are also more and more recognized. As a chiral pesticide with enantiomers, compared with the inefficiency and high environmental pollution of racemic pesticides,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P7/42C12R1/66
CPCC12N1/14C12P7/42C12N1/145C12R2001/66
Inventor 薛亚平周海岩胡海峰姜瑞王银龙李一作郑裕国王远山
Owner ZHEJIANG UNIV OF TECH
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