A kind of method utilizing lipase to prepare chitosan oligosaccharide

A technology of lipase and chitosan oligosaccharide, which is applied in the field of preparing chitosan oligosaccharide by using lipase, can solve the problems of color influence of degradation products, long reaction time, and many kinds of enzymes, so as to achieve good product quality and uniform color. , the effect of short enzymatic hydrolysis time

Active Publication Date: 2019-09-03
GUANGDONG PHARMA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Chinese patent document CN 1320123 C discloses a method for producing low-molecular-weight chitosan by using lipase. The enzymolysis reaction is carried out under the conditions of a substrate concentration of 10-100 g / L and a reaction time of 5-6 hours, and the filtrate is vacuum-concentrated and filtered. Gel chromatographic column to obtain low molecular weight chitosan, this method effectively reduces the production cost, but the required reaction time is long, the enzyme liquid and the air contact time are long and easy to be oxidized, which inevitably affects the degradation products in the later stage. color makes a big difference
Tan Peiyi and others studied the orthogonal experiment method to optimize the process of preparing chitosan oligosaccharides by lipase. The optimal conditions determined were: the amount of lipase added was 1.2g / 100mL, the enzymatic hydrolysis temperature was 50°C, the pH of the solution was 4.5, and the enzymatic hydrolysis The time is 6h, but the time required for the reaction is long and the yield is low
Chinese patent document CN 102492664 B discloses a method for preparing chitosan oligosaccharides using compound enzymes, wherein the compound enzymes include cellulase, lysozyme, amylase, lipase, glucose and papain. rate, the obtained chitosan oligosaccharide products are evenly distributed, but there are many types of enzymes required, and the steps are complicated
Therefore, it is necessary to overcome the problems of low yield, long reaction time and complicated steps caused by the high viscosity of the current chitosan substrate, so as to provide people with a new type of simple operation and short reaction time using lipase to prepare chitosan. sugar method

Method used

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  • A kind of method utilizing lipase to prepare chitosan oligosaccharide
  • A kind of method utilizing lipase to prepare chitosan oligosaccharide
  • A kind of method utilizing lipase to prepare chitosan oligosaccharide

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Embodiment 1

[0028] Embodiment 1, the present invention utilizes lipase to prepare the method for chitosan oligosaccharide

[0029] S1 dissolves: chitosan is dissolved in the acetic acid-sodium acetate buffer solution of pH value 4.0 and adds 0.1% sucrose fatty acid ester (HLB value 10) of chitosan weight, is made into the chitosan that mass concentration is 1.0% sugar solution;

[0030] S2 lipase hydrolysis: add a lipase solution with a concentration of 0.5% to the chitosan solution obtained in step S1, so that the mass ratio of the enzyme to the bottom is 20%, and place it in a constant temperature water bath at 40°C for 0.5h to obtain the enzymolysis solution, take it out and boil it in a water bath 10 minutes for inactivation, cooled to room temperature;

[0031] S3 adjusting pH and dialysis: adding dropwise NaOH solution with a mass concentration of 5% to the enzymolysis solution obtained in step S2 to adjust the pH value to 7.0-8.0, so that water-insoluble chitosan is precipitated, ...

Embodiment 2

[0034] Embodiment 2, the present invention utilizes lipase to prepare the method for chitosan oligosaccharide

[0035] S1 dissolves: chitosan is dissolved in the acetic acid-sodium acetate buffer solution of pH value 4.5 and adds 0.4% sucrose fatty acid ester (HLB value 12) of chitosan weight, is made into the chitosan that mass concentration is 1.7% sugar solution;

[0036] S2 lipase hydrolysis: add the lipase liquid that concentration is 0.5% to the chitosan solution gained in step S1, make the mass ratio of enzyme bottom be 22%, 45 ℃ constant temperature water baths 1h, obtain enzymolysis liquid, after taking out, boiling water bath 10 Minutes for inactivation, cooled to room temperature;

[0037]S3 adjusts pH and dialysis: in the enzymolysis solution gained in step S2, add dropwise the NaOH solution that mass concentration is 5% to adjust pH to 7.0~8.0, make water-insoluble chitosan separate out, take filtrate after filtering and discard filter residue, will The filtrate...

Embodiment 3

[0040] Embodiment 3, the present invention utilizes lipase to prepare the method for chitosan oligosaccharide

[0041] S1 dissolves: chitosan is dissolved in the acetic acid-sodium acetate buffer solution of pH value 5.0 and adds the sucrose fatty acid ester (HLB value 13) of 0.5% chitosan weight, is made into the chitosan that mass concentration is 2.0% sugar solution;

[0042] S2 lipase hydrolysis: add lipase liquid with a concentration of 0.5% to the chitosan solution obtained in step S1, so that the mass ratio of the enzyme to the bottom is 30%, 50 ℃ constant temperature water bath for 1.5h, to obtain the enzymolysis solution, take it out and boil it in water bath 10 minutes for inactivation, cooled to room temperature;

[0043] S3 adjusts pH and dialyzes: in the enzymolysis solution gained in step S2, add dropwise the solution that mass concentration is 5% NaOH to adjust pH to 7.0~8.0, make water-insoluble chitosan separate out, take filtrate after filtering and discard ...

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Abstract

Provided is a method for preparing chitosan oligosaccharide by using a lipase. The method comprises dissolving chitosan in an acetic acid-sodium acetate buffer solution and adding sucrose fatty acid ester to prepare a chitosan solution, adding a lipase to the chitosan solution for enzymatic hydrolysis, dialyzing an enzymatic hydrolysate after the enzymatic hydrolysis, and concentrating the resulting dialysate and drying.

Description

technical field [0001] The invention belongs to the technical field of oligosaccharide preparation, and in particular relates to a method for preparing chitosan oligosaccharide by using lipase. Background technique [0002] Chitosan, also known as deacetylated chitin, is a long-chain macromolecule. It has poor water solubility under physiological conditions and high solution viscosity. Its application as a biological effector is very limited. Oligochitosan is a low-molecular-weight polymer with a low degree of polymerization (2-20) obtained by degrading chitosan. Its segment molecules are connected with a large number of reactive groups hydroxyl and amino groups, so it has a low molecular weight. , high solubility, good moisture absorption and moisturizing properties, strong absorption capacity and good biocompatibility, etc., and also has anti-inflammatory, enhanced immunity, lower cholesterol, lower blood fat, anti-tumor, antibacterial, anti-virus, clear Physiological ac...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/26
CPCC12P19/26
Inventor 苏政权曹华
Owner GUANGDONG PHARMA UNIV
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