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A strain of Escherichia coli jl-glcn and its application

A technology of Escherichia coli, CGMCCNO.13924, applied in the direction of bacteria, microorganisms, microorganisms, etc., can solve the problems of long enzymatic hydrolysis process, poor efficiency, incomplete hydrolysis, etc.

Active Publication Date: 2020-07-07
HENAN JULONG BIOLOGICAL ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The direct enzymatic hydrolysis method means that the production of N-acetylglucosamine can be produced by directly enzymolyzing chitin with chitinase. Due to the particularity of the chemical properties of chitin, the enzymatic hydrolysis process is long and incomplete, and the hydrolysis rate is 75%. Below %, the efficiency is low, the actual large-scale industrial production cost is high, and the benefit is not good

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  • A strain of Escherichia coli jl-glcn and its application
  • A strain of Escherichia coli jl-glcn and its application
  • A strain of Escherichia coli jl-glcn and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Escherichia coli JL-GlcN and common N-acetylglucosamine-producing Escherichia coli were respectively inoculated on the slant, cultured in a biochemical incubator at 37°C for 20 hours, and then diluted in a gradient and spread on the complete medium (LB). Pick a single colony and inoculate it on the slant, and incubate at 30-36°C for about 12 hours in a biochemical incubator. Inoculate into 500mL seed bottles, culture conditions: temperature 30-36°C, shaker speed 200rmp, culture time 6-8 h; when the OD value grows to about 1.0-2.0, inoculate into 500ml fermentation bottles, inoculum size: 7 %, culture conditions: temperature 30-36 ℃, shaker speed 200rmp, when the cell OD is about 10-15, add IPTG induction, and continue to culture for 10-20h. After the fermentation, the N-acetylglucosamine content was detected to be 60.26g / L and 52.38g / L. The yield of N-acetylglucosamine increased by 15.04%.

Embodiment 2

[0035] Pick one ring of Escherichia coli JL-GlcN and ordinary N-acetylglucosamine-producing Escherichia coli from fresh slant surfaces and inoculate them into 500mL seed bottles. The cultivation time is 6-8 hours; when the OD value grows to about 1.0-4.0, put the shake flask seeds into a 5L automatic fermenter with 3L fermentation medium at an inoculation amount of 10% for secondary seed cultivation. The culture temperature is 30-36℃, the tank pressure is 0.01-0.05MPa, the air flow rate is 1-8L / min and the stirring speed is 200-800r / min to maintain the dissolved oxygen at 10-50%, and the pH value is controlled between 7.0-7.2. The time is 5-8h. The secondary seeds were inserted into a 30L fully automatic fermenter equipped with 18L fermentation medium with 10% inoculum to carry out enzyme production fermentation. The fermentation temperature is 30-36°C, the tank pressure is 0.01-0.15MPa, the air flow rate is 5-50L / min and the stirring speed is 200-800r / min to maintain the dis...

Embodiment 3

[0037] Pick one ring of Escherichia coli JL-GlcN and ordinary N-acetylglucosamine-producing Escherichia coli from fresh slant surfaces and inoculate them into 500mL seed bottles. The culture time is 6-8h; when the OD value grows to about 1.0-4.0, insert the shake flask seeds into the 0.6m 3 2m of fermentation medium 3 Secondary seed cultures were carried out in fermenters. Cultivation temperature 30-36℃, tank pressure 0.01-0.05MPa, air flow 0.1-0.5m 3 / min and stirring speed 100-300r / min to maintain the dissolved oxygen at 10-50%, the pH value is controlled between 7.0-7.2, and the culture time is 5-8h. Insert the secondary seeds with 10% inoculum into the 6m 3 10m of fermentation medium 3 Enzyme fermentation is carried out in the fermenter. Fermentation temperature 30-36℃, tank pressure 0.01-0.15MPa, air flow 0.5-6m 3 / min and stirring speed 100-300r / min to maintain the dissolved oxygen at 10-40%, the fermentation pH value is controlled between 7.0-7.2, and when the cel...

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Abstract

The present invention relates to a strain of Escherichia coli JL-GlcN and its application. The Escherichia coli JL-GlcN has a strain preservation number of CGMCC NO.13924; the Escherichia coli JL-GlcN is activated on a slope, Incubate in a biochemical incubator at 37°C for 20 hours, then apply gradient dilution on LB medium, pick a single colony and inoculate them on the LB slant medium, and incubate in a biochemical incubator at 30-36°C for 12 hours; Cultivate in a fermenter with a fermentation medium. When the cell OD is 10-15, add IPTG to induce, cultivate for 10-20 h, and obtain a fermentation liquid containing N-acetylglucosamine; extract the fermented liquid containing N-acetylglucosamine N‑acetylglucosamine in solution. The fermentation of the Escherichia coli JL-GlcN bacterial strain of the present invention is aerobic fermentation, and the bacterial body grows fast; the fermentation process is simple, easy to control, and easy to scale up for industrial production.

Description

technical field [0001] The invention relates to an Escherichia coli strain, specifically an Escherichia coli JL-GlcN and application thereof. Background technique [0002] N-acetylglucosamine is the monomer of chitin, and the stock of chitin in nature is very large, second only to cellulose. N-acetylglucosamine has important physiological functions in medicine, and has anti-inflammatory, anti-tumor and anti-oxidative effects. application. [0003] The existing preparation methods of N-acetylglucosamine include sodium methoxide method and direct enzymatic hydrolysis of chitin. The process of the sodium methoxide method is as follows: add sodium methoxide to the raw material of glucosamine for replacement reaction, remove chloride ions in glucosamine, then add acetic anhydride to acetylate it, and remove bacteria, rust, colloid and other harmful substances in it through ultrafiltration , followed by fractional precipitation and filtration, followed by redissolution crystall...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P19/26C12R1/19
CPCC12P19/26C12N1/205C12R2001/19
Inventor 曹华杰刘帅王卫富谢沛刘晓东
Owner HENAN JULONG BIOLOGICAL ENG CO LTD