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Production method and application of high-efficient cellulase mixture

A technology of cellulase and production method, which is applied in the field of fermentation engineering, can solve the problems of slow degradation rate of solid substrate, carbohydrate repression, high cost of medium, etc., and achieve the goal of overcoming carbohydrate repression, increasing production capacity, and reducing repression effect Effect

Pending Publication Date: 2016-03-23
SHANDONG LONGLIVE BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Patent CN200910091968 provides a method for high-yield cellulase, through feed and dissolved oxygen to control the concentration of bacteria in the fermentation process to improve the activity of cellulase fermentation enzyme fermentation, the feed is very complex ingredients, including expensive Sophorose, lactose, etc. and a variety of trace elements, while the cost of the medium is relatively expensive
How to not only solve the problem of rapid nutrition demand and slow degradation rate of solid substrate in the late stage of liquid submerged fermentation, but also solve the problem of easy carbohydrate repression caused by adding glucose is a difficult problem faced by those skilled in the art

Method used

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  • Production method and application of high-efficient cellulase mixture
  • Production method and application of high-efficient cellulase mixture
  • Production method and application of high-efficient cellulase mixture

Examples

Experimental program
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Effect test

Embodiment 1

[0032] 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.

[0033] 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.

[0034] Feed medium: 30% xylose mother liquor, 0.5% (w / v) magnesium sulfate, 0.005% (w / v) zinc sulfate, its composition of xylose mother liquor comprises 50% (w / v) xylose, 10% ( w / v) glucose, 10% (w / v) arabinose, balance water.

[0035] Penicillium oxalicum JUA10-1 (gifted by Shandong University) was inoculated in the bran medium by streaking at 28-32°C for 7 days, and after the pink spores grew, it was transferred to fresh bran medium and cultured for 5 days as the fungus. kind. Connect 1cm ...

Embodiment 2

[0040] 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.

[0041] Enzyme production medium: 6.5% 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.

[0042] Feed medium: 30% glucose mother solution, 0.6% (w / v) magnesium sulfate and 0.003% (w / v) zinc sulfate; wherein the glucose content in the glucose mother solution is 50%, the disaccharide content is 20%, and the polysaccharide content is 5% .

[0043] Streak inoculation of Trichoderma reesei (China Center for Microbial Culture Collection, collection number CICC40932) in bran medium for 7 days at 28-32°C, and transfer to fresh bran medium for culture after green spores grow. 5 days as...

Embodiment 3

[0059] Crush the straw to 1-3cm, solid-liquid ratio 1:8, 2% sodium hydroxide at 80°C and stir for 1.5h, wash with water to pH 7.0, adjust pH to 4.8 with sulfuric acid, and saccharify with 10% substrate In the experiment, the cellulase enzyme liquid described in the examples and comparative examples was used to carry out the saccharification experiment according to the enzyme activity of 5, 7.5, and 10 U per g of slag. Among them, in example 2 and comparative example 2, β was also added according to the amount of substrate -Glucosidase 3IU / g, measure glucose and calculate conversion rate, the result is as shown in table 3 and table 4, the result shows that adopting the cellulase enzyme liquid enzyme activity prepared by the present invention not only obviously improves, and conversion efficiency is also higher than equivalent enzyme The conversion rate of straw slag in alkaline pretreatment was increased by more than 15%, and the optimal enzyme dosage was reduced from 10U / g slag...

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Abstract

The invention relates to a production method and application of a high-efficient cellulase mixture. Through performing fermentation by using a non-soluble carbon source, not only can fermentation cost of cellulase be saved, but also the production of the cellulase is effectively induced; a reducing sugar control measure is adopted, so that the repression effect of a carbohydrate is reduced, and the production capacity of the cellulase is improved; in addition, a novel material-making up control manner is built through combining the non-soluble carbon source with a material-making up component and a material-making up rate process, so that matching of all components of the produced cellulase mixture is optimized while the yield (production capacity) of the cellulase is promoted, and the activity of the cellulase is improved by not less than 50 percent.

Description

technical field [0001] The invention relates to the technical field of fermentation engineering, in particular to a production method and application of a high-efficiency cellulase mixture. 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 extracellula...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12P19/14C12R1/885C12R1/80
CPCC12N9/2437C12P19/14
Inventor 肖林夏蕊蕊覃树林杨建李红震孙保剑
Owner SHANDONG LONGLIVE BIO TECH CO LTD
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