Alpha-glucosidase immobilization method

A technology of glucosidase and chitosan, which is applied in the direction of immobilization on/in the organic carrier, can solve the problems of large loss of enzyme activity, unstable properties, no immobilization of α-glucosidase, etc., and achieve revival High recovery and reduced loss of enzyme activity

Inactive Publication Date: 2005-05-25
SHANGHAI UNIV
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Problems solved by technology

But this cross-linking reagent has some following problems: in the cross-linking reaction, the C=N Schiff bond formed by the aldehyde group and the amine group is easy to hydrolyze, especially at a higher temperature, the hydrolysis is more serious, and the enzyme activity is lost during the immobilization process. Larger; many different substances are formed due to automatic polymerization during storage, and the properties are unstable; in addition, glutaraldehyde is also a toxic substance, which is harmful to the operator
According to reports, the enzymes immobilized with the trihydroxymethylphosphine cross-linking reagent include urease, alcohol dehydrogenase, etc., but there is no report on the use of this reagent to immobilize α-glucosidase

Method used

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  • Alpha-glucosidase immobilization method

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Embodiment 1

[0015] Embodiment one: the specific process steps of the present embodiment are as follows:

[0016] a. Pretreatment of chitosan: first prepare chitosan with high degree of deacetylation, get 20g chitosan, add 350mL concentration of 40% NaOH, stir reaction time at 130°C for 5 hours, then wash with water until neutral, Dried at 105°C, pulverized, and passed through a 120-mesh sieve to obtain pretreated chitosan with a deacetylation degree of 93%.

[0017] b. Preparation of trihydroxymethylphosphorus (THP): take equimolar amounts of tetrakishydroxymethylphosphorus chloride (THPC) and KOH, dissolve them in 90mL and 10mL of water respectively, and dissolve the KOH aqueous solution at room temperature under vigorous stirring Add it dropwise to THPC aqueous solution, and the reaction ends when the dropwise addition is completed. The concentration of trihydroxymethylphosphine (THP) obtained in this reaction solution was 2.5 mg / ml.

[0018] c. Cross-linking agent to immobilize α-glu...

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Abstract

A process for immobilizing alpha-glucosidase by using chitosan as carrier and using trihydroxymethyl phosphorus for cross-linking includes removing acetyl from chitosan, reacting between phoshporus tetrahydroxymethyl chloride and KOH to obtain trihydroxymethyl phosphorus, adding the deacetylated chitosan, reacting, filtering, water wasing, washing by potassium phosphate solution as buffering liquid, adding alpha-glucosidase, immobilizing, washing by water and then said buffering liquid, and storing at 4 deg.C.

Description

technical field [0001] The invention relates to a process method for immobilizing α-glucosidase, which belongs to the technical field of organic chemical enzyme chemical engineering. Background technique [0002] According to reports, Japan, Europe, the United States and China are all concerned about the industrial production and related research of isomalto-oligosaccharides. As the key enzyme preparation for the industrial production of isomalt-oligosaccharides, α-glucosidase has attracted much attention from the food industry at home and abroad. However, due to the poor stability and controllability of α-glucosidase, as well as the high price, the development and application of isomaltooligosaccharides are restricted to a certain extent. The main content of enzyme engineering is to immobilize free enzymes, called immobilized enzymes. The use of immobilized enzyme technology can not only reduce the cost, but also improve the acid and alkali resistance stability, thermal st...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/04C12N11/10
Inventor 雍克岚吕敬慈卢大胜陈旭冯偲慜顾慧娟
Owner SHANGHAI UNIV
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