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A kind of method and application of solid-state fermentation for preparing heat- and acid-tolerant glucose oxidase

A technology of glucose oxidase and solid-state fermentation, which is applied in the field of microorganisms, can solve problems such as hindering widespread application, and achieve the effects of promoting bacterial growth and enzyme synthesis, preventing premature aging, and enhancing stability

Active Publication Date: 2020-07-17
西宝生物科技(上海)股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, glucose oxidase has been used in actual production as a flour and feed additive, but there are still technical problems hindering its wide application

Method used

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  • A kind of method and application of solid-state fermentation for preparing heat- and acid-tolerant glucose oxidase
  • A kind of method and application of solid-state fermentation for preparing heat- and acid-tolerant glucose oxidase
  • A kind of method and application of solid-state fermentation for preparing heat- and acid-tolerant glucose oxidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Step 1, preparation of strains: first, under aseptic conditions, pick the test tube spores of Paecilomyces thermophilic species stored in two rings and inoculate them on the activated slant medium (potato extract (100g potatoes cut into 1cm 3 Cube, boiled for 30min, filtered with 4 layers of gauze, supplemented with water to 100mL) 100mL, bran extract (15g of bran, added with water 100mL, boiled for 30min, filtered to obtain filtrate, supplemented with water to 100mL) 100mL, inorganic salt solution (take (NH4 ) 2 SO 4 : 5g, KH 2 PO 4 : 5g, MgSO 4 ·7H 2 0: 1g, NaNO 3 : 1g, CaCl 2 : 1g, FeCl 3 : 1g, Tween 80: 5mL) 1mL, sucrose 1g, agar powder 3g, pH5.0, add water to 1000ml) to activate; place the activated slant medium in a 30-35℃ incubator for 3-5 days; then , inject 1mL of sterile saline into the activated Paecilomyces thermophila slant medium, pipette the spores on the slant medium repeatedly, and further dilute the spore concentration to 10 7 1 / mL, then take ...

Embodiment 2

[0040] Step 1, preparation of strains: first, under aseptic conditions, pick the test tube spores of Paecilomyces thermophilic species stored in two rings and inoculate them on the activated slant medium (potato extract (100g potatoes cut into 1cm 3 Cube, boiled for 30min, filtered with 4 layers of gauze, supplemented with water to 100mL) 100mL, bran extract (15g of bran, added with water 100mL, boiled for 30min, filtered to obtain filtrate, supplemented with water to 100mL) 100mL, inorganic salt solution (take (NH4 ) 2 SO 4 : 8g, KH 2 PO 4 : 8g, MgSO 4 ·7H 2 0: 1.5g, NaNO 3 : 3g, CaCl 2 : 3g, FeCl 3 : 1.5g, Tween 80: 8mL) 1mL, sucrose 1g, agar powder 3g, pH5.5, add water to 1000ml) for activation; place the activated slant medium in a 33°C incubator for 4 days; then, to activate Inject 1mL of sterile saline into the good Paecilomyces thermophila slant medium, and repeatedly blow the spores on the slant medium with a pipette gun to further dilute the spore concentratio...

Embodiment 3

[0046] Step 1, preparation of strains: first, under aseptic conditions, pick the test tube spores of Paecilomyces thermophilic species stored in two rings and inoculate them on the activated slant medium (potato extract (100g potatoes cut into 1cm 3 Cube, boiled for 30min, filtered with 4 layers of gauze, supplemented with water to 100mL) 100mL, bran extract (15g of bran, added with water 100mL, boiled for 30min, filtered to obtain filtrate, supplemented with water to 100mL) 100mL, inorganic salt solution (take (NH4 ) 2 SO 4 : 10g, KH 2 PO 4 : 10g, MgSO 4 ·7H 2 0: 2g, NaNO 3 : 5g, CaCl 2 : 5g, FeCl 3 : 2g, Tween 80: 10mL) 1mL, sucrose 1g, agar powder 3g, pH6.0, add water to 1000ml) to activate; place the activated slant medium in a 35°C incubator for 3-5 days; then, add Inject 1 mL of sterile saline into the activated Paecilomyces thermophila slant medium, pipette the spores on the slant medium repeatedly, and further dilute the spore concentration to 10 7 1 / mL, then ...

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Abstract

The invention discloses a method of preparing heat-resistant acid-resistant glucose oxidase via solid fermentation and application of heat-resistant acid-resistant glucose oxidase, and belongs to the technical field of microorganisms. The method comprises the steps of 1, preparing a bacterial strain; 2, performing flask solid fermentation of Paecilomyces thermophila; 3, performing koji tray solid fermentation of Paecilomyces thermophila; 4, preparing a glucose oxidase preparation. Liquid nutrient material is supplemented periodically during solid fermentation of Paecilomyces thermophila, and fermentation parameters are adjusted and periodically controlled during fermentation according to growth characteristics and enzyme-producing characteristics of bacteria; compared with common solid fermentation methods, the method produces the glucose oxidase that has activity increased by about 45% and that has good stability and high acid tolerance; fermentation medium materials are low in price and mainly include common agricultural byproducts, bran, corn stalk powder, corn powder and soybean meal, and waste media from Cordyceps militaris cultivation, and the cultivation cost is low.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a method for preparing glucose oxidase by solid-state fermentation of a heat-resistant and acid-resistant glucose oxidase-producing Paecilomyces thermophilic strain and an application thereof. Background technique [0002] Glucose oxidase (Glucose oxidase, referred to as GOD) can highly specifically catalyze the reaction between β-D-glucose and oxygen in the air, oxidizing glucose into 1,5-gluconolactone and hydrogen peroxide, and then gluconate Lactone combines spontaneously with water to produce gluconic acid. Due to the unique properties and safety of the enzyme, it has shown high application value in food, medicine, feed and other industries. In terms of food, glucose oxidase can be used as a flour improver to improve the tissue structure of gluten, improve the air holding capacity, elasticity, and water holding capacity of dough; because glucose oxidase c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/04A23K50/75A23K20/189C12R1/79
CPCC12N9/0006C12Y101/03004
Inventor 高兆建张铁柱孙会刚刘恩岐李同祥曹建冬顾强沈彬彬
Owner 西宝生物科技(上海)股份有限公司
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