Method of immobilizing beta-fructofuranosidase and glucose oxidase by sol-gel process

A technology of fructofuranosidase and glucose oxidase, which is applied in the field of enzyme engineering, can solve the problems of inability to immobilize other enzymes, decrease in enzyme activity, and application limitations, and achieve improved yield of lactulose oligosaccharides, good stability, and low cost Effect

Active Publication Date: 2018-10-16
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the ordinary sol-gel method for the immobilization of β-fructofuranosidase will inhibit the release of the enzyme activity embedded in the gel, and the enzyme a

Method used

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  • Method of immobilizing beta-fructofuranosidase and glucose oxidase by sol-gel process
  • Method of immobilizing beta-fructofuranosidase and glucose oxidase by sol-gel process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] First activate the Arthrobacter sp.10137 strain, insert the preserved Arthrobacter into the activation medium (glucose 2% (w / v); yeast extract 0.15% (w / v); magnesium sulfate 0.01% (w / v); Potassium dihydrogen phosphate 0.2% (w / v); diammonium hydrogen phosphate 0.6% (w / v); pH7.0-7.2), cultured at 30° C. for 24 hours to obtain activated strains. Carry out seed culture afterwards, draw the bacterial classification after 1ml activation to house 100ml seed culture medium (glucose 2% (w / v); Yeast extract 0.15% (w / v); Peptone 0.3% (w / v); Magnesium sulfate 0.01% (w / v); Potassium dihydrogen phosphate 0.2% (w / v); Diammonium hydrogen phosphate 0.6% (w / v), pH7.0-7.2) in a 250ml Erlenmeyer flask, in a shaker for 30 Cultivate at ℃ for 24h, with a shaking speed of 125r / min. Carry out fermentation culture then, seed culture solution is connected with 100ml fermentation medium (sucrose 4% (w / v); Yeast extract 1.2% (w / v): peptone 0.8% (w / v) by 2% inoculum size access ); Magnesium sulfat...

Embodiment 2

[0050] First activate the Arthrobacter sp.10137 strain, insert the preserved Arthrobacter into the activation medium (glucose 2% (w / v); yeast extract 0.15% (w / v); magnesium sulfate 0.01% (w / v); Potassium dihydrogen phosphate 0.2% (w / v); diammonium hydrogen phosphate 0.6% (w / v); pH7.0-7.2), cultured at 30° C. for 24 hours to obtain activated strains. Carry out seed culture afterwards, draw the bacterial classification after 1ml activation to house 100ml seed culture medium (glucose 2% (w / v); Yeast extract 0.15% (w / v); Peptone 0.3% (w / v); Magnesium sulfate 0.01% (w / v); Potassium dihydrogen phosphate 0.2% (w / v); Diammonium hydrogen phosphate 0.6% (w / v), pH7.0-7.2) in a 250ml Erlenmeyer flask, in a shaker for 30 Cultivate at ℃ for 24h, with a shaking speed of 125r / min. Carry out fermented culture then, seed culture liquid is connected with 100ml fermentation medium (sucrose 4% (w / v); Yeast extract 1.2% (w / v): peptone 0.8% (w / v) by 3% inoculum size access ); Magnesium sulfate 0.2...

Embodiment 3

[0054] First activate the Arthrobacter sp.10137 strain, insert the preserved Arthrobacter into the activation medium (glucose 2% (w / v); yeast extract 0.15% (w / v); magnesium sulfate 0.01% (w / v); Potassium dihydrogen phosphate 0.2% (w / v); diammonium hydrogen phosphate 0.6% (w / v); pH7.0-7.2), cultured at 30° C. for 24 hours to obtain activated strains. Carry out seed culture afterwards, draw the bacterial classification after 1ml activation to house 100ml seed culture medium (glucose 2% (w / v); Yeast extract 0.15% (w / v); Peptone 0.3% (w / v); Magnesium sulfate 0.01% (w / v); Potassium dihydrogen phosphate 0.2% (w / v); Diammonium hydrogen phosphate 0.6% (w / v), pH7.0-7.2) in a 250ml Erlenmeyer flask, in a shaker for 30 Cultivate at ℃ for 24h, with a shaking speed of 125r / min. Carry out fermented culture then, seed culture solution is connected with 100ml fermentation medium (sucrose 4% (w / v); Yeast extract 1.2% (w / v): peptone 0.8% (w / v) by 4% inoculum size access ); Magnesium sulfate 0...

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Abstract

The application provides a method of immobilizing beta-fructofuranosidase and glucose oxidase by a sol-gel process. The method comprises the steps of (1) producing beta-fructofuranosidase; (2) isolating and purifying beta-fructofuranosidase; (3) immobilizing the purified beta-fructofuranosidase and glucose oxidase solution to gel in the presence of a silane coupling agent. Compared with traditional production of lactosucrose via immobilization of beta-fructofuranosidase, the method has the advantages that the sol-gel process is used herein for the first time to immobilize beta-fructofuranosidase and glucose oxidase so as to improve enzymic stability, high enzyme activity recovery rate is attained, lactosucrose yield is increased, and the method is simpler to operate than the traditional art, has mild conditions and is an environment-friendly method.

Description

technical field [0001] The invention relates to the technical field of enzyme engineering, in particular to a new method for producing lactulose-oligosaccharides through co-immobilization of β-fructofuranosidase and glucose oxidase by a sol-gel method. Background technique [0002] β-fructofuranosidase (EC 3.2.1.26) is a glycosidase that widely exists in plants and microorganisms. It not only has hydrolysis, can catalyze the hydrolysis of sucrose into glucose and fructose, but also has transglycosylation, and can be widely used Used in the synthesis of functional oligosaccharides, food additives, modification and modification of glycosides and other biologically active substances. However, the properties of β-fructofuranosidases from different sources are different, and the ability of transglycosylation and receptor specificity are also different. As a raw material, the fructosyl produced by the hydrolysis of sucrose is transferred to the C at the reducing end of lactose 1...

Claims

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Application Information

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IPC IPC(8): C12N11/08C12N9/04C12N9/26
CPCC12N9/0006C12N9/2431C12N11/08C12Y101/03004C12Y302/01026
Inventor 金征宇龙杰潘婷徐学明谢正军周星柏玉香焦爱权田耀旗赵建伟王金鹏
Owner JIANGNAN UNIV
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