Oxidation resistant low-temperature glucose oxidase and production method and application of oxidation resistant low-temperature glucose oxidase

A technology of glucose oxidase and oxidation resistance, which is applied in the field of bioengineering and can solve problems such as increased production costs

Active Publication Date: 2017-12-19
SHANDONG LONGKETE ENZYME PREPARATION
View PDF3 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, almost all glucose oxidase depends on imports, and the production cost also increases accordingly.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Oxidation resistant low-temperature glucose oxidase and production method and application of oxidation resistant low-temperature glucose oxidase
  • Oxidation resistant low-temperature glucose oxidase and production method and application of oxidation resistant low-temperature glucose oxidase
  • Oxidation resistant low-temperature glucose oxidase and production method and application of oxidation resistant low-temperature glucose oxidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Mutation breeding of embodiment 1 bacterial strain

[0037] Preparation of spore suspension: elute the spores on the slant of the original strain with an appropriate amount of sterile physiological saline, place them in a pre-sterilized triangular flask with glass beads, vibrate on a shaker for 20 minutes, and filter them out with sterilized absorbent cotton. Mycelium to dispersed single spore suspension, counted with a hemocytometer. diluted to 10 8 individual / mL spore suspension.

[0038] Microwave mutagenesis: put a test tube containing 5 mL of spore suspension in a beaker containing ice, use a microwave oven with a frequency of 2450 MHz and an output power of 700 W, irradiate the test tubes one by one at different times, and the dilution gradient is 10 -1 ~10 -6 . Take 10 of each dose -4 ~10 -6 Spread 0.2 mL of spore suspension at 3 dilutions on the plate medium, incubate at 30°C for 2-3 days, count the number of colonies, and draw the lethality curve. Pick ...

Embodiment 2

[0046] Example 2 Production of glucose oxidase by liquid fermentation of bacterial strain CH870 and its extraction

[0047] (1) Seed tank cultivation:

[0048] Tank pressure 0.05-0.08MPa, cultivation temperature 30°C, ventilation volume 30m 3 / h, the stirring speed is 180r / min, and the pH is controlled to 7.0; cultivate until the thalli are darkly stained, strong, and free of bacteria, and the seed liquid is obtained after the cultivation is completed;

[0049] Seed tank medium: glucose 20%, peptone 25%, (NH 4 ) 2 SO 4 5%, K 2 HPO 4 1%, MgSO 4 ·7H 2 O0.5%, pH 7.0;

[0050] (2) Fermentation tank culture

[0051] Inoculate the seed liquid into the fermenter according to 5% inoculation amount, the tank pressure is 0.05-0.08Mpa, the culture temperature is 30°C, the stirring speed is 300r / min, the pH is controlled at 6.5-7.0; the ventilation rate: 0-40h is 30m 3 / h, 40-58h is 45m 3 / h, 58h - 40m for the tank 3 / h; when the pH rises to 7.0, start feeding and control th...

Embodiment 3

[0067] Example 3 Glucose oxidase enzyme activity assay method

[0068] Substrate system: pipette 2 mL of 0.07 g / L o-dianisidine solution and 1 mL of 5% glucose solution in a graduated test tube with a 1 mL pipette gun. Use a 0.5mL pipette to pipette 0.1mL0.1g / L horseradish peroxidase solution into the same graduated test tube. The substrate system was placed in a constant temperature water bath at 30 °C for 10 min.

[0069] Determination of enzyme activity: Use a pipette gun to draw 0.1mL of the diluted enzyme solution and shake it evenly in the substrate. Using a blank tube as a control, quickly measure the absorbance at 460nm with a visible spectrophotometer. Read the initial absorbance value as A 0 And timing, record the absorbance value A every 1min n , a total of 5min was measured.

[0070] From the measured absorbance value, calculate the enzyme activity of the enzyme solution according to the following formula: X1 (U / mL)

[0071] ΔA n+1 =A n+1 -A n (n=0,1,2,3,4)...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention belongs to the technical field of bioengineering, and particularly relates to a low-temperature glucose oxidase and a production method and application of the low-temperature glucose oxidase. The glucose oxidase is produced from Aspergillu niger CH870, and the preservation number is CGMCC No.14138. The enzyme activity of a fermentation liquid of the glucose oxidase produced through fermentation of a strain reaches more than 2300 U/ml, the produced glucose oxidase is stable when the pH is within 2.0-8.0, the most appropriate reaction temperature is 20 DEG C, the stability is good, the glucose oxidase resists oxidation, the residual enzyme activity is 100% under the condition of 30 mmol hydrogen peroxide, the residual enzyme activity is 90% under the condition of 200 mmol hydrogen peroxide, and the application is extensive.

Description

Technical field: [0001] The invention belongs to the technical field of bioengineering, in particular to a low-temperature glucose oxidase with improved oxidation resistance, a production method and application thereof. Background technique: [0002] Glucose oxidase (GOD) can specifically oxidize β-D-glucose into gluconic acid and hydrogen peroxide. Glucose oxidase has a wide range of applications in the fields of food, medicine and biology. [0003] Because glucose oxidase can catalyze glucose to consume oxygen and produce gluconic acid and hydrogen peroxide, it is widely used in the food industry, mainly in the following four aspects: removal of residual glucose in food, deoxygenation, sterilization, determination of food glucose content. [0004] Glucose oxidase, as a new type of green enzyme preparation, was listed as one of the 12 permitted feed additives by the state in 1999. The mechanism of action of glucose oxidase to promote growth is: the glucose oxidase added i...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/04C12N1/14C12R1/685
CPCC12N1/14C12N9/0006C12Y101/03004C12N1/145C12R2001/685
Inventor 王兴吉贾仁洁钱娟娟郭庆文张杰
Owner SHANDONG LONGKETE ENZYME PREPARATION
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap