Arthrobacter nicotinovorans WYG001 and application thereof in preparation of N-BOC-L-homoserine lactone

A technology of Arthrobacter nicotinophilus and homoserine lactone, which is applied in the field of acetin lactone, can solve problems such as difficulty in industrial production and long reaction time, and achieve low environmental pollution, strong regional selectivity, and low energy consumption Effect

Active Publication Date: 2015-04-22
ZHEJIANG UNIV OF TECH
View PDF3 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The screened microbial strain Arthrobacter nicotinovorans Strain stereoselectively catalyzes racemized homoserine in phosphate solution to obtain optically pure enantiomers e.e. S >99.9%, the yield of D configuration is 25%, and the yield of homoserine of L-configuration is about 4%, and the conversion time is 48h, the reaction time is too long, and it is not easy for industrial production

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Arthrobacter nicotinovorans WYG001 and application thereof in preparation of N-BOC-L-homoserine lactone
  • Arthrobacter nicotinovorans WYG001 and application thereof in preparation of N-BOC-L-homoserine lactone
  • Arthrobacter nicotinovorans WYG001 and application thereof in preparation of N-BOC-L-homoserine lactone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 1. Screening of microorganisms

[0037] The microorganisms involved in the present invention are screened and separated by the following procedures: Weigh 1 to 2 g of wet soil samples (obtained from soil under different environmental conditions across the country) and suspend them in 10 mL of 0.5% sterile saline, shake and mix; ~2mL of suspension in 30mL of liquid enrichment medium, cultured at 30°C and 200r / min for 2 to 3 days, then aseptically transfer 1mL of turbid bacterial solution to fresh liquid enrichment medium for continuous enrichment 3 times. In the enrichment medium, N-BOC-DL-homoserine lactone is used as the sole carbon source, and the formula is as follows: NaNO 3 4.0g / L,KH 2 PO 4 2.0g / L, MgSO 4 ·7H 2 O 0.5g / L, KCl 1.0g / L, ZnSO 4 0.05g / L, the solvent is water, pH 7.0. The concentration of N-BOC-DL-homoserine lactone enriched in the first round was 1 g / L, the concentration in the second round was 3 g / L, the concentration in the third round was 5 g / L...

Embodiment 2

[0046] Slant culture: Arthrobacter nicotinophilus WYG001 was inoculated into the slant medium, and cultured at 30° C. for 24 hours to obtain the slant of the bacteria; the composition of the slant medium was the same as in Example 1.

[0047] Seed culture: Pick an inoculation loop of bacteria from the slant and inoculate it into 100 mL of seed culture medium, cultivate at 30°C and 200 r / min for 24 hours to obtain seed liquid; the composition of the seed culture medium is the same as that in Example 1.

[0048] Fermentation culture: Under sterile conditions, inoculate 10mL of seed liquid into 150mL of enzyme-producing medium, culture at 30°C and 200r / min for 24 hours to obtain a fermentation liquid, and centrifuge the fermentation liquid at 12,000rpm for 10 minutes after the cultivation, discard the supernatant , to obtain Arthrobacter nicotinophilus WYG001 wet bacteria. Wet cells were taken in an amount of 1.2 g / ml and added to pH 7.0, 0.1 mol / L phosphate buffer, stirred evenl...

Embodiment 3

[0050] Take 0.34g of wet bacteria obtained by the method in Example 2, add 50mg of N-BOC-DL-homoserine lactone and 5mL of 0.1M, pH7.0 phosphate buffer, and stir for 7.5h at 30°C and 200rpm. The reaction solution was filtered to remove the enzyme, and the filtrate was analyzed by gas chromatography to determine the content of converted N-Boc-DL-homoserine lactone, and the enzyme activity was calculated. The specific enzyme activity of the fermentation broth was 36.85 U / L.

[0051] Enzyme activity is defined as: U is 1 enzyme activity unit, and the 1 enzyme activity unit is the hydrolysis of 1 μmol of N-Boc-homoserine lactone into N-Boc-homoserine per minute under specified conditions. the amount of enzyme required.

[0052] Gas chromatographic analysis conditions: gas chromatograph Agilent 6890 type gas chromatograph; chiral gas chromatographic column BGB-175 (0.25mm × 30m); carrier gas N 2 (14.54kPa), split ratio 20:1, inlet temperature 250°C, detector temperature 220°C; oven...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses arthrobacter nicotinovorans WYG001 and application thereof in preparation of N-BOC-L-homoserine lactone. The N-BOC-L-homoserine lactone is prepared by taking a wet bacterium body obtained from the fermentation cultivation of the arthrobacter nicotinovorans WYG001 or a dry bacterium body obtained by freeze-drying the wet bacterium body as a catalyst, N-BOC-DL-homoserine lactone as a substrate and 0.1M of a phosphate buffer solution at pH7.0 as a reaction medium through carrying out a conversion reaction at 20-50 DEG C, and separating and purifying a reaction liquid after the reaction, so as to obtain the N-BOC-L-homoserine lactone. The method disclosed by the invention is strong in enzyme regioselectivity, high in reaction conversion rate, simple in downstream separation, low in energy consumption, slight in environmental pollution and suitable for industrial production.

Description

(1) Technical field [0001] The present invention relates to a synthesis method of N-BOC-L-homoserine lactone, in particular to the use of Arthrobacter nicotinophilus WYG001 (CCTCC M 2014054) to catalyze racemic BOC-DL-homoserine lactone for stereoselective hydrolysis A method for synthesizing optically pure N-BOC-L-homoserine lactone by reaction, and then synthesizing L-selenomethionine from N-BOC-L-homoserine lactone. (2) Background technology [0002] L-selenomethionine, English name: L-(+)-Selenomethionine, is an important animal food additive amino acid, and is also an important intermediate for preparing other selenium-containing drugs. Its main functions are: it can enhance the sperm motility of animals; it can increase the body's antioxidant capacity and immunity; it has anti-cancer and anti-cancer effects; [0003] Homoserine lactone in optically pure form is an important chiral intermediate widely used in the fields of medicine and food additives. In the invention...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N1/20C12P13/06C12R1/06
CPCC12P7/62C12P13/12C12P41/003C12N1/205C12R2001/06
Inventor 王宇光朱冰春吴冬梅
Owner ZHEJIANG UNIV OF TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap