Efficient production method for cellulase

A technology of cellulase and streak inoculation, applied in the field of fermentation engineering

Pending Publication Date: 2016-01-20
SHANDONG LONGLIVE BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the prior art has achieved certain results, how to solve the carbon water repression effect (CCR) in the cellulase fermentation process and improve the cellulose production capacity is still the main problem faced by those skilled in the art

Method used

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  • Efficient production method for cellulase
  • Efficient production method for cellulase
  • Efficient production method for cellulase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Seed medium: 1% xylose residue, 1% bran, 1% glucose, 1% peptone, 0.2% ammonium sulfate, 0.3% potassium dihydrogen phosphate, 0.05% magnesium sulfate, 50mL in a 500mL Erlenmeyer flask, extinguished at 121°C Bacteria 20min.

[0042] Enzyme production medium: 3% xylose residue, 3% bran, 0.2% microcrystalline cellulose, 0.5% bean cake powder, 0.2% ammonium sulfate, 0.1% urea, 0.3% potassium dihydrogen phosphate, 0.05% magnesium sulfate, 0.3 % Tween 80, sterilized at 121°C for 30min.

[0043] Filter paper enzyme (FPA) enzyme activity determination method: the fermentation broth was centrifuged at 8000rpm for 10min, and the supernatant was taken as crude enzyme solution for the determination of enzyme activity and protein content. Refer to the light industry industry standard (QB2583-2003) for determination, take 0.5mL of appropriately diluted crude enzyme solution, use 50mg (1cm×6cm) Xinhua filter paper strips as substrate, add 1.5mL and 0.5mL of pH4.8 acetate buffer, Taki...

Embodiment 2

[0048] Seed medium: 1% xylose residue, 1% bran, 1% glucose, 1% peptone, 0.2% ammonium sulfate, 0.3% potassium dihydrogen phosphate, 0.05% magnesium sulfate, 50mL in a 500mL Erlenmeyer flask, extinguished at 121°C Bacteria 20min.

[0049] Enzyme production medium: 6% xylose residue, 4% bran, 0.2% microcrystalline cellulose, 0.5% bean cake powder, 0.2% ammonium sulfate, 0.1% urea, 0.3% potassium dihydrogen phosphate, 0.05% magnesium sulfate, 0.3 % Tween 80, sterilized at 121°C for 30min.

[0050] The filter paper enzyme (FPA) enzyme activity assay method and Bradford assay protein content were as described above.

[0051] Penicillium oxalicum JUA10-1 was streak-inoculated in bran medium and cultured at 30°C for 5 days. After the pink spores grew, it was transferred to fresh bran medium and cultured for 5 days as the strain. Connect 2cm 2 The strains were inoculated in the seed medium, fermented at 28-32°C and 200rpm for 24-48h, and prepared into first-grade seeds, and inocula...

Embodiment 3

[0054] Seed medium: 1% xylose residue, 1% bran, 1% glucose, 1% peptone, 0.2% ammonium sulfate, 0.3% potassium dihydrogen phosphate, 0.05% magnesium sulfate, 50mL in a 500mL Erlenmeyer flask, extinguished at 121°C Bacteria 20min.

[0055] Enzyme production medium: 6% xylose residue, 4% bran, 0.2% microcrystalline cellulose, 0.5% bean cake powder, 0.2% ammonium sulfate, 0.1% urea, 0.3% potassium dihydrogen phosphate, 0.05% magnesium sulfate, 0.3 % Tween 80, sterilized at 121°C for 30min.

[0056] The filter paper enzyme (FPA) enzyme activity assay method and Bradford assay protein content were as described above.

[0057] Penicillium oxalicum JUA10-1 was streak-inoculated in bran medium and cultured at 30°C for 5 days. After the pink spores grew, it was transferred to fresh bran medium and cultured for 5 days as the strain. Connect 10cm 2 The strain is inoculated in the seed medium, fermented at 28-32°C and 200rpm for 24-48h, and prepared into first-grade seeds, inoculated wi...

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Abstract

The invention relates to an efficient production method for cellulase, and particularly discloses a method for controlling cellulase fermentation through the oxygen uptake rate. Through optimal screening of culture media and optimal controlling over oxygen uptake rate conditions, the stable fermentation method is determined, and the enzyme activity of a produced cellulase preparation is improved by more than 50 percent; the technology can be widely applied to cellulase production through filamentous fungi such as penicillium and aspergillus.

Description

technical field [0001] The invention relates to the technical field of fermentation engineering, in particular to a cellulase production method. Background technique [0002] Cellulase has a wide range of sources and complex components. It is widely found in the metabolites of insects, molluscs, fungi, bacteria, and actinomycetes. The main production route is microbial fermentation. Among them, filamentous fungi and bacterial cellulase It is the most widely used, and currently mainly uses fungi to ferment and produce cellulase. [0003] Among bacteria, most aerobic bacteria and anaerobic bacteria are rich in cellulase, especially the anaerobic bacteria in the rumen that are rich in a variety of cellulose-degrading enzymes. 20% of the cellulase in the world cellulose market comes from Trichoderma and Aspergillus. Mainly because filamentous fungi have many advantages of producing enzymes: ①The cellulase produced is an extracellular enzyme, which is convenient for the separat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42
CPCC12N9/2437C12N9/2445C12Y302/01004C12Y302/01021C12Y302/01091
Inventor 程少博肖林夏蕊蕊杨建覃树林李红震孙保剑
Owner SHANDONG LONGLIVE BIO TECH CO LTD
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