Method for preparing chitosan oligosaccharides by feed-batch-oxidative pretreatment assistant with enzyme hydrolysis of chitosan oligosaccharides

A fed-batch and pretreatment technology, applied in fermentation and other directions, can solve the problems of low production efficiency, low substrate concentration, high viscosity, etc., and achieve the effects of less chemical reagents, mild reaction conditions, and high product safety

Pending Publication Date: 2018-09-28
XIAMEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to overcome the deficiencies such as high viscosity, low substrate concentration and low production efficiency in the existing

Method used

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  • Method for preparing chitosan oligosaccharides by feed-batch-oxidative pretreatment assistant with enzyme hydrolysis of chitosan oligosaccharides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Disperse 3g of chitosan uniformly in 96mL of water, add 1mL of glacial acetic acid, stir to dissolve at room temperature, then add 4mL of 30% hydrogen peroxide, stir at 60℃ for 5min, and quickly add 1mL of glacial acetic acid and 3g of chitosan to control the total pretreatment For 1h, add 0.2g catalase to remove hydrogen peroxide, adjust the pH with concentrated ammonia, add 5g cellulase, and react at 55℃ for 12h. The obtained degradation solution is adjusted with ammonia to adjust the pH to 7, using 10kDa ultra The enzyme is removed from the filter membrane, and the filtrate is vacuum freeze-dried to obtain a pale yellow chito-oligosaccharide product, in which the yield of the chito-oligosaccharide that can be dissolved in 75% ethanol reaches 90%. For product identification by thin layer chromatography, see figure 1 (a), S is the standard product, and 1 to 4 are the same samples. Repeat 4 times. The 7 points of the standard from top to bottom are GlcN: Glucosamine; (G...

Embodiment 2

[0024] According to Example 1, 1 mL of glacial acetic acid and 3 g of chitosan were added at the 5th and 15th min of pretreatment, the total pretreatment time was controlled for 0.5h, 0.2g of catalase was added to remove hydrogen peroxide, and the pH was adjusted with concentrated ammonia. Add 5g of cellulase and react in a shaker at 55°C for 6 hours. The product can be dissolved at pH 7-14. Continue to react for 12 hours. The obtained degradation solution is adjusted to pH 7 with ammonia, and the enzyme is removed by a 10kDa ultrafiltration membrane. The filtrate is vacuum freeze-dried to obtain a light yellow chito-oligosaccharide product, in which the yield of chito-oligosaccharide that can be dissolved in 75% ethanol reaches 70%. For product identification by thin layer chromatography, see figure 1 (b). S is the standard product; 5-8 are the same samples repeated 4 times. The 7 points of the standard from top to bottom are GlcN: Glucosamine; (GlcN) 2 :Chitobiose; (GlcN) 3...

Embodiment 3

[0026] According to Example 2, add 1.2g NaHSO during enzymolysis 3 After 12 hours of enzymatic hydrolysis, almost pure white chitooligosaccharides were obtained. Among them, the yield of chitooligosaccharides that can be dissolved in 75% ethanol reached 48%.

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Abstract

The invention discloses a method for preparing chitosan oligosaccharides by feed-batch-oxidative pretreatment assistant with enzyme hydrolysis of chitosan oligosaccharides, and relates to a method forpreparing low-molecular-weight chitosan. The method comprises the steps that degradable solution is obtained through feed-batch-oxidative pretreatment of chitosan oligosaccharides; the hydrogen peroxide is removed; enzymatic hydrolysis and inhibition of side reactions of the chitosan is carried out; and product separation and drying is carried out, and the chitosan oligosaccharides is obtained. In the process oxidative pretreatment, the substrate concentration in a chitosan degradation system is increased the feed-batch method, the initial hydrolysis is achieved, and finally, the chitosan wasfurther hydrolyzed by the enzyme. Compared with the high-concentration hydrolysis of the chitosan, the pretreatment method has simple equipment and high adaptability, can easily be combined with a enzyme membrane reactor, immobilized enzyme and other technologies, and has a great application prospect in the preparation of the chitosan oligosaccharides with narrow molecular weight distribution.

Description

Technical field [0001] The present invention relates to a preparation method of low molecular weight chitosan, in particular to a method for preparing chitosan oligosaccharides by assisting enzymatic hydrolysis of chitosan in batch feed-oxidation pretreatment. Background technique [0002] Chitosan is a linear macromolecule connected by glucosamine and acetylglucosamine through β-1,4 glycosidic bonds. Its molecular weight ranges from hundreds of thousands to millions. It is the only naturally occurring positively charged polymer in nature. Sugar is insoluble in neutral and alkaline solutions, nor can it be dissolved in organic solvents. Although it is soluble in acidic aqueous solutions, it has limited solubility and extremely high viscosity, which seriously affects the application of chitosan. Low-molecular-weight chitosan is an incomplete hydrolysis product of chitosan, which retains the basic properties of chitosan such as adsorption, positive charge, reducibility, and alkalin...

Claims

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Application Information

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IPC IPC(8): C12P19/26C12P19/14
CPCC12P19/14C12P19/26
Inventor 刘健李蒙龙敏南甘礼惠
Owner XIAMEN UNIV
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