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Fermentation production method for N-acetylglucosamine

An acetamido and production method technology, applied in the field of fermentation engineering, can solve problems such as low conversion rate, allergic reaction, unstable raw material source, etc., and achieve the effects of improving content and conversion rate, and optimizing flow acceleration rate

Inactive Publication Date: 2015-10-21
ANHUI BBCA FERMENTATION TECH ENG RES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, most of the glucosamine in my country is produced from chitosan, but there are shortcomings such as unstable raw material sources, and some people have allergic reactions.
Although microbial fermentation produces N-acetylglucosamine (in the fermented liquid that contains N-acetylglucosamine, add the hydrochloric acid of 0.1mol / L, react 3h under 100 ℃, can convert more than 90% of N-acetylglucosamine into Glucosamine) can overcome the above shortcoming, but there are low output, low conversion rate and high content of by-product acetic acid and glutamic acid in the production of N-acetylglucosamine by microbial fermentation. Therefore, it is necessary to develop a fermentation of N-acetylglucosamine A new process to increase the fermentation yield and conversion rate of N-acetylglucosamine while reducing the content of fermentation by-products acetic acid and glutamic acid

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] (1) Preparation of seed medium: glucose 15g / L, dipotassium hydrogen phosphate 11g / L, potassium dihydrogen phosphate 16g / L, yeast extract powder 1g / L, threonine 0.2g / L, methionine 0.15g / L, Magnesium sulfate 0.5g / L, ammonium sulfate 5g / L, manganese sulfate 0.013mg / L, iron sulfate 0.024mg / L, zinc sulfate 0.047mg / L, cobalt chloride 0.01mg / L.

[0032] Put it into a 50L fermenter, adjust the pH of the seed medium to 7.3 with ammonia water with a concentration of 25% (m / m), pass saturated steam to the seed medium to 121°C, keep it warm for 30min, and set the seed culture condition: temperature 37 ℃, tank pressure 0.05mpa, ventilation ratio 1:0.5, speed 300rpm, pH6.8-7.0. The N-acetylglucosamine-producing Escherichia coli is inserted for cultivation, and when the seed OD660nm=3.5-5, it is used as a seed maturity standard.

[0033] (2) Preparation of fermentation medium: glucose 5g / L (single elimination), dipotassium hydrogen phosphate 16g / L, potassium dihydrogen phosphate 10g / ...

Embodiment 2

[0039] When the residual sugar drops below 0.5g / L during the N-acetylglucosamine fermentation and cultivation process, add 55% (m / m) glucose solution to the N-acetylglucosamine fermentation broth according to the flow acceleration rate of 6g / L h . At this time, add urea solution at a rate of 10:1 mass ratio between glucose solution and urea solution, that is, add 15% (m / m) urea solution to N-acetylglucosamine fermentation broth according to the flow acceleration rate of 0.6g / L h . Other conditions and culture methods are the same as in Example 1. When fermented for 54 hours, the content of N-acetylglucosamine was 110g / L, the conversion rate was 40.1%, the content of acetic acid was 1.9g / L, and the content of glutamic acid was 2.1g / L.

Embodiment 3

[0041] When the residual sugar drops below 0.5g / L during the N-acetylglucosamine fermentation and cultivation process, add 55% (m / m) glucose solution to the N-acetylglucosamine fermentation broth according to the flow acceleration rate of 7g / L h . At this time, add urea solution at a rate of 10:1 mass ratio between glucose solution and urea solution, that is, add 15% (m / m) urea solution to N-acetylglucosamine fermentation broth according to the flow acceleration rate of 0.7g / L h . Other conditions and culture methods are the same as in Example 1. When fermented for 54 hours, the content of N-acetylglucosamine was 98g / L, the conversion rate was 38.5%, the content of acetic acid was 3.2g / L, and the content of glutamic acid was 3.5g / L.

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Abstract

The invention provides a fermentation production method for N-acetylglucosamine. In the fermentation process, when the residual glucose content of fermentation liquor falls below 0.5 g / L, a glucose solution is flowingly added into the fermentation liquor at the flow acceleration of 5-8 g / L.h, when OD660nm of the fermentation liquor is equal to 28-31, IPTG is added for induction, and then a urea solution is flowingly added into the fermentation liquor at the flow acceleration of 0.5-0.8 g / L.h. According to the fermentation production method for the N-acetylglucosamine, in the fermentation process, induction is performed through the IPTG, the flow adding speed of the glucose solution and the urea solution is optimized when induction is performed, and the N-acetylglucosamine content and the conversion rate at the fermentation end point are significantly improved.

Description

technical field [0001] The invention relates to the field of fermentation engineering, in particular to a fermentation production method of N-acetylglucosamine. Background technique [0002] Glucosamine has a wide range of uses, mainly used in food, medical, cosmetics, agriculture and animal husbandry and other fields. At present, glucosamine is mostly used for the prevention and treatment of arthritis. Glucosamine is the main component of connective tissue and chondrocytes. If joint disease occurs, supplementing glucosamine can repair damaged cartilage and increase the lubrication between joints. [0003] At present, my country's glucosamine is mostly produced with chitosan as raw material, but there are shortcomings such as unstable source of raw material, and some people have allergic reactions. Although microbial fermentation produces N-acetylglucosamine (in the fermented liquid that contains N-acetylglucosamine, add the hydrochloric acid of 0.1mol / L, react 3h under 10...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/26
Inventor 张家泉杨为华徐斌张雪锋
Owner ANHUI BBCA FERMENTATION TECH ENG RES
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