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High-activity cellulase and its preparation method

A cellulase and manufacturing method technology, applied in the directions of biochemical equipment and methods, enzymes, enzymes, etc., can solve the problems of difficult large-scale industrial production, unsatisfactory effect, affecting popularization and application, etc., to improve quality and efficiency, The effect of facilitating large-scale industrial production and reducing production costs

Inactive Publication Date: 2007-02-28
ACAD OF STATE GRAIN ADMINISTRATION
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AI Technical Summary

Problems solved by technology

Solid fermentation equipment requires less investment, but covers a large area, is prone to contamination by bacteria, and has poor production stability. Moreover, the technology is not easy to master, and it is difficult to carry out large-scale industrial production. At present, most enterprises in my country still use solid fermentation technology to produce cellulase
Liquid fermentation has high production stability, easy to master technology, and is convenient for large-scale industrial production. However, because cellulase is an inducing enzyme, natural cellulose substances such as corn stalks, wheat straw, and rice straw are used as inducers, and the effect is not very satisfactory. Therefore, most enterprises use expensive cellulose as raw material in cellulase liquid fermentation, which greatly increases the production cost and affects the popularization and application of this technology.

Method used

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  • High-activity cellulase and its preparation method

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Effect test

Embodiment 1

[0029] Implementation example 1. Screen and isolate high-activity cellulase-producing bacteria from Hebei cotton field soil samples

[0030] Separation medium: 0.35% CMC (carboxymethyl cellulose), 0.05% MgSO 4 ·7H 2 O, 0.1%KH 2 PO 4 , 1%Na 2 CO 3 (separately sterilized), 1% NaNO 3 , 1% NaCl, 1.8% agar was added for solidification.

[0031] Suspend the soil sample in a sterile 0.85% saline solution, dilute it step by step under sterile conditions, then spread the diluted sample suspension on a hydroxymethylcellulose (CMC)-agar medium, and incubate at about 30°C After 3 days, the strains were initially screened, and the range of colonies capable of producing cellulase produced hydrolysis circles, which were shown as cool areas. The diameter of the cool areas was the measure of CMC enzyme activity, and the strains with larger diameters were selected as rough screening strains.

[0032] Compound medium for enzyme production: bran 1%, cellulose 2%, ammonium sulfate 0.3%, pep...

Embodiment 2-7

[0039] Example 2-7. Preparation of highly active cellulase

[0040] The basidiomycete CGMCCNo.1294 was used as the production strain, and the medium with the following ratio was used, and the liquid strain was sterilized and inserted into the Erlenmeyer flask for liquid submerged fermentation. The medium formula and reaction conditions are shown in Table 1.

[0041] The carbon source is cotton, cotton linters or cotton textile scraps, and the length of the cotton fiber is less than 1cm, or 1-2% hydrochloric acid solution is used, heated and hydrolyzed at 80-90°C for 8-10 hours, and neutralized to a pH of 6.

[0042] The filling capacity of the liquid medium in the fermenter is 70%, the pH value of the medium is 4.0-6.0, the fermentation temperature is 28-30°C, the ventilation rate is 1:0.04-1:1.0, and the fermentation time is 96-120 hours; The fermented liquid is filtered through a plate frame to remove mycelium, the clarified fermented liquid is concentrated 8-10 times in a vac...

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Abstract

The present invention relates to one kind of high activity cellulase and its preparation process, and is especially one kind high activity cellulase prepared with basidiomycetes and its preparation process in microbe preparation. Basidiomycetes CGMCC No. 1294 as the producing strain is inoculated into culture medium with carbon source, wheat bran, bean cake powder, ammonium sulfate, potassium dihydrogen phosphate, magnesium sulfate and Tween-80 in certain proportion for deep liquid fermentation in an Erlenmeyer flask in certain conditions for 96-120 hr; and the fermented liquid is filtered, concentrated and spray dried to obtain the cellulase powder. The present invention has the advantages of stable production process, less foreign bacteria infection and being suitable for industrial production. The produced cellulase has enzyme activity as high as 40000 u / g.

Description

technical field [0001] The invention relates to a high-activity cellulase and a production method thereof, in particular to a high-activity cellulase produced by basidiomycetes and a production method thereof, belonging to the field of microorganism production. Background technique [0002] Cellulase is widely used in food, feed, brewing, textile and other fields, and can be divided into fungal-derived cellulase and bacterial-derived cellulase according to its source. At present, the microbial strains used to produce cellulase are mostly filamentous fungi, and the strains with strong enzyme activity are Trichoderma, Aspergillus and Penicillium. In particular, Trichoder mavirde and its related strains are more typical, and are currently recognized as better cellulase-producing bacteria. Ruminants rely on rumen microorganisms to digest cellulose, such as rumen bacteria (Rumenococcus xanthophyllum, Ruminococcus albicans, etc.) and Clostridium thermocellum, etc., but the yield ...

Claims

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Application Information

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IPC IPC(8): C12N9/42C12N1/14
Inventor 程树峰唐芳伍松陵张晓琳
Owner ACAD OF STATE GRAIN ADMINISTRATION
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