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Preparation method of Pichia pastoris beta-glucosidase preparation

A technology of glucosidase and Pichia pastoris, which is applied in the field of preparation of Pichia pastoris β-glucosidase preparation, can solve the problems of low yield, low protease activity, and few types of production strains, etc., and achieve low production cost , low impurity content, high enzyme activity effect

Inactive Publication Date: 2019-02-01
石河子开发区天易特色果蔬加工生产力促进中心(有限责任公司)
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are many β-glucosidases in the market, the types of production strains used are less, and the output of the β-glucosidases of the existing bacterial strains producing β-glucosidases is not large, and the protease produced Enzyme activity is not high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] The technical scheme that this specific embodiment adopts is: the concrete steps of a kind of preparation method of Pichia pastoris β-glucosidase preparation are as follows: A, adopt ultraviolet light combined with laser composite mutagenesis Pichia saccharomyces: (a), from Pichia pastoris Pick a single colony of Pichia kluyveri from the solid plate of Red Saccharomyces, inoculate it in the yeast extract powder old glucose medium for cultivation and fermentation and obtain the fermentation broth: 50ml yeast extract powder peptone glucose medium, in Cultivate at 30°C for 40 hours at a speed of 150 rpm; centrifuge the fermentation broth at a speed of 4000 r / min for 10 minutes, discard the supernatant, and dilute with sterile water to obtain a bacterial solution with a concentration of 200 cells / ml; draw Put 10ml of bacterial solution on a sterile plate and place it under ultraviolet light irradiation, place it equidistantly, and 30 cm away from the ultraviolet light, and i...

Embodiment 2

[0018]The difference from Example 1 is that its specific steps are as follows: A, using ultraviolet light combined with laser compound mutagenesis of Pichia: (a), picking a single Pichia kluyveri from the solid plate of Pichia Bacterial colonies were inoculated in yeast extract powder old glucose medium for cultivation and fermentation to obtain fermentation broth: 50ml of yeast extract powder peptone glucose medium was cultivated at 30°C for 48 hours at a speed of 300 rpm; Centrifuge the fermentation broth at a speed of 5000r / min for 10min, discard the supernatant, and dilute it with sterile water to obtain a bacterial solution with a concentration of 200 cells / ml; draw 10ml of the bacterial solution onto a sterile plate and place it under ultraviolet light, and place it equidistantly , and 30 centimeters away from the ultraviolet lamp, irradiated for 30 minutes to obtain the first mutagenic strain. Then carry out He-Ne laser compound mutagenesis (632.8nm, 11.5mW), carry out ...

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Abstract

The invention relates to the field of biological food engineering, in particular to a preparation method of a Pichia pastoris beta-glucosidase preparation, comprising A, inducing Pichia pastoris by the combination of ultraviolet and laser; B, preparing a Pichia kluyveri enzyme preparation. The preparation method according to the technical scheme has the advantages that the production cost is low,the steps of ethanol precipitation, centrifuging, salting-out, ultrafiltration, lyophilization and the like are used in the preparation process, the product has low impurity content and meets the requirements of the food additive standard, the Pichia pastoris beta-glucosidase preparation with high enzymic activity is prepared, and the yield of beta-glucosidase preparations is increased.

Description

technical field [0001] The invention relates to the fields of biology and food engineering, in particular to a preparation method of Pichia pastoris β-glucosidase preparation. Background technique [0002] β-glucosidase (β-glucosidase, EC 3.2.1.21, referred to as BG) is a hydrolytic enzyme, which belongs to cellulase and is an important component of cellulolytic enzyme system. At present, β-glucosidase is widely used in food, feed and medical fields, etc., and has a good development prospect. In terms of food, β-glucosidase can hydrolyze the flavor precursor glycosides in fruits and release volatile aglycone, which can enhance the aroma of wine and other fruit wines and fruit juices; it can also hydrolyze soybean isoflavones to produce highly active soybeans Isoflavone aglycon products; can also add amygdalin, thereby reducing the bitterness. Making it play an important role in scientific research and industrial application is of great significance to human production and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12N1/16C12N15/01C12R1/84
CPCC12N1/16C12N9/2445C12N15/01C12Y302/01021
Inventor 孙静涛史学伟王军武强李小军
Owner 石河子开发区天易特色果蔬加工生产力促进中心(有限责任公司)
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