Method for producing N-acetyl glucosamine by enzymatically degrading chitin

An acetamido and enzymatic degradation technology, applied in the field of enzymatic degradation of chitin to produce N-acetylglucosamine, can solve the problems of high cost of gel adsorption method, difficult control of conversion process, large loss of enzymatic activity, etc. The effect of low production time, low cost and high conversion efficiency

Inactive Publication Date: 2015-03-04
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional enzyme concentration methods include salt precipitation, reverse osmosis and other methods, and there are many problems: the loss of enzyme activity during the salt precipitation process is cumbersome, and the operation is cumbersome; the gel adsorption method is expensive and difficult to promote
In addition, the degradation products of chitin can be used as both carbon source and nitrogen source utilized by bacteria, the conversion process is difficult to control, and it is very easy to infect bacteria.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Step 1, microbial fermentation produces chitinase

[0033] select Chitinolyticbacter meiyuanensis The SYBC-H1 strain was used as a seed and inoculated into the medium (2 g / L glucose, 2 g / L peptone, 0.7 g / L KH 2 PO 4 , 0.3 g / L K 2 HPO 4 ·3H 2 O, 0.4 g / LMgSO 4 ·7H 2 O, pH 7.0), cultured at 37°C and 200rpm for 12 h, and then inoculated with an enzyme-producing fermentation medium (3 g / L powdered chitin, 3 g / L Jerusalem artichoke powder, 3 g / L urea, 0.7 g / L KH 2 PO 4 , 0.3 g / L K 2 HPO 4 3H2O, 0.5 g / LMgSO 4 ·7H 2 O) A 7.5 L fermenter with a liquid volume of 4.5 L was fermented at a temperature of 26 °C, a rotation speed of 250 rpm, an aeration volume of 1 vvm, and an initial pH of 7.5 for 72 h before collecting the fermentation broth.

[0034] Step 2, the acquisition of crude enzyme solution

[0035] Centrifuge the fermented liquid obtained in step 1 at 8000g in a centrifuge at 4°C, separate the bacteria, collect the supernatant to obtain a crude chiti...

Embodiment 2

[0042] Step 1, microbial fermentation produces chitinase

[0043] select Chitinolyticbacter meiyuanensis The SYBC-H1 strain was used as a seed and inoculated into the medium (4 g / L glucose, 4 g / L peptone, 0.7 g / L KH 2 PO 4 , 0.3 g / L K 2 HPO 4 ·3H 2 O, 0.5 g / LMgSO 4 ·7H 2 O, pH 7.0), cultured at 37°C and 200 rpm for 12 h, and then inoculated with an enzyme-producing fermentation medium (4 g / L powdered chitin, 4 g / L Jerusalem artichoke powder 4 g / L, urea 0.7 g / LKH 2 PO 4 , 0.3 g / L K 2 HPO 4 ·3H 2 O, 0.5 g / LMgSO 4 ·7H 2 O) A 7.5L fermenter with a liquid capacity of 4.5L was fermented at a temperature of 26°C, a rotation speed of 250 rpm, and an aeration volume of 2vvm for 72 hours at an initial pH of 7.5, and the fermented liquid was collected after fermentation.

[0044] Step 2, the acquisition of crude enzyme solution

[0045] Centrifuge the fermented liquid obtained in step 1 at 8000g in a centrifuge at 4°C, separate the bacteria, collect the supernatant to obt...

Embodiment 3

[0052] Step 1, microbial fermentation produces chitinase

[0053] select Chitinolyticbacter meiyuanensis The SYBC-H1 strain was used as a seed and inoculated into the medium (4 g / L glucose, 4 g / L peptone, 0.7 g / L KH 2 PO 4 , 0.3 g / L K 2 HPO 4 ·3H 2 O, 0.5 g / LMgSO 4 ·7H 2 O, pH 7.0), cultured at 37°C and 200 rpm for 12 h, and then inoculated with an enzyme-producing fermentation medium (4 g / L powdered chitin, 4 g / L Jerusalem artichoke powder 4 g / L, urea 0.7 g / LKH 2 PO 4 , 0.3 g / L K 2 HPO 4 ·3H 2 O, 0.5 g / LMgSO 4 ·7H 2 O) A 7.5L fermenter with a liquid volume of 4.5L was collected at a temperature of 26°C, a rotation speed of 250 rpm, an aeration volume of 2vvm, and an initial pH of 7.5 for 72 hours.

[0054] Step 2, the acquisition of crude enzyme solution

[0055] Centrifuge the fermented liquid obtained in step 1 at 8000g in a centrifuge at 4°C, separate the bacteria, collect the supernatant to obtain a crude chitinase enzyme liquid, store it at 4°C, and use i...

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PUM

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Abstract

The invention discloses a method for producing N-acetyl glucosamine by enzymatically degrading chitin. The method comprises the following steps: step 1, producing chitinase by microbial fermentation; step 2, obtaining a crude enzyme solution; step 3, concentrating the enzyme solution; step 4, producing N-acetyl glucosamine. The invention firstly provides the method for producing N-acetyl glucosamine by enzymatically degrading chitin, and an enzyme production medium is low in cost and short in production cycle. The material used in the step of concentrating the enzyme solution can be recycled, the operation process is easy, and the enzyme recovery is high. A measure of preventing bacterial contamination is adopted in the degradation process, so that the purpose that the N-acetyl glucosamine with high medical value is efficiently and quickly produced by using cheap chitin with low value as a raw material is achieved, and the method has significant economic and social benefits.

Description

technical field [0001] The invention relates to a method for enzymatically degrading chitin to produce N-acetylglucosamine. [0002] Background technique [0003] Chitin (C 8 h 13 o 5 N)n, also known as chitin, chitin or chitin, is a polymer composed of N-acetylglucosamine through glycosidic bonds. important ingredient. The molecular structure is a long chain polysaccharide composed of 8000 monomers, which is a translucent and strong material in nature. Chitin can be degraded into chitosan oligosaccharides and monosaccharides by chemical methods and biological enzymatic methods. Currently, the commonly used chemical degradation methods have disadvantages such as huge environmental pollution, low reaction efficiency, poor product quality, and difficult process control, and have been gradually adopted. Bioenzymatic degradation of chitin replacement. [0004] N-acetylglucosamine is the latest third-generation health-care functional food additive in chitin health food ser...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/26C12N9/42C12R1/01
Inventor 陈可泉张阿磊高聪何珣欧阳平凯
Owner NANJING UNIV OF TECH
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