Method for preparing cellulase by bacilli

A technology of bacillus and cellulase, which is applied in the field of biochemistry to achieve the effects of short production cycle, high activity and good stability

Active Publication Date: 2012-11-07
ZHONGRONG TECH CORP LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a gap in domestic production of alkaline cellulase

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: Erlenmeyer flask fermentation prepares cellulase

[0033] 1. Slant culture: under aseptic conditions, use an inoculation loop (2.5mm inner diameter, 10 microliters, the same below) to pick a bacillus (Bacillus Cohn, preservation number CGMCC No.6211) strain and insert it into the test tube In the slant medium, culture at 35°C for 3 days; the mass percentage of each component in the slant medium is: soluble starch 0.5% peptone 1.0% yeast powder 1.0% sodium chloride 0.5% dipotassium hydrogen phosphate 0.1%, CMC (carboxymethyl Sodium cellulose) 0.5%, sodium carbonate 1%, agar 2.0%; the rest is water.

[0034] 2. Oscillating culture: under aseptic conditions, pick two loops of the slant strains into the seed medium of the Erlenmeyer flask, place on a shaker for 36 hours, culture temperature is 36°C, shaker speed is 150r / min. The mass percent of each component in the seed medium is: soluble starch 0.5% peptone 1.0% yeast powder 1.0% bran 0.5% sodium chloride...

Embodiment 2

[0038] Embodiment 2: 10L fermentation tank prepares cellulase

[0039]1. Slant culture: Under aseptic conditions, pick a Bacillus Cohn strain (CGMCC No. 6211) with an inoculation loop and insert it into the slant medium of the test tube, and cultivate it at 37°C for 3 days; the slant medium The mass percentage of each component in the product is: soluble starch 0.3%, peptone 0.8%, yeast powder 0.8%, sodium chloride 0.4%, dipotassium hydrogen phosphate 0.1%, CMC 0.5%, sodium carbonate 1%, agar 2.0%, and the rest for water.

[0040] 2. Oscillating culture: under aseptic conditions, pick two loops of the slant strains into the seed medium of the Erlenmeyer flask, place on a shaker for 36 hours, culture temperature is 36°C, shaker speed is 150r / min. The mass percentage of each component in the seed medium is: soluble starch 0.4%, peptone 1.0%, yeast powder 0.8%, bran 0.5%, sodium chloride 0.4%, dipotassium hydrogen phosphate 0.1%, sodium carbonate 0.6%, and the rest is water. ...

Embodiment 3

[0044] Example 3: 30m 3 Preparation of cellulase in fermenter

[0045] 1. Slant culture: Under aseptic conditions, pick one of the Bacillus Cohn (CGMCC No.6211) strains with an inoculation loop and insert them into the slant medium of the test tube, and cultivate them at 35-37°C for 3-4 days The mass percent of each component in the slant medium is: soluble starch 0.8%, peptone 1.2%, yeast powder 1.2%, sodium chloride 0.6%, dipotassium hydrogen phosphate 0.2%, CMC 0.6%, sodium carbonate 1.2%, agar 2.0%, the rest is water.

[0046] 2. Oscillating culture: under aseptic conditions, put the slant strains into the seed medium of the Erlenmeyer flask with an inoculation ring, put them on a shaker and shake them for 36 hours, the culture temperature is 37°C, and the shaker speed is 150r / h min. The mass percentage of each component in the seed medium is: soluble starch 0.6%, peptone 1.2%, yeast powder 1.2%, bran 0.8%, sodium chloride 0.3%, dipotassium hydrogen phosphate 0.1%, sodi...

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PUM

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Abstract

The invention provides a method for preparing cellulase by bacilli. The method includes the steps of using bacillus as a producing strain, subjecting the producing strain to slant culture, shaking culture and fermenting culture to obtain the cellulase. Fermenting culture medium comprises, by weight percent, 0.5-2% of starch, 1-5% of soybean cake meal, 8-12% of soybean cake meal hydrolysate, 0.3-0.8% of bran, 1-2% of inorganic salt, and 0-0.3% of foaming agent. The cellulase prepared by the method has high activity in neutral pH and alkali conditions and is better than acidic cellulase in terms of water abrasion enzyme application. Starch material is used as a carbon source by the method, and accordingly the cellulase is better than that of the fiber material in terms of transfer, feeding, sterilization and the like. The soybean cake meal hydrolysate is used as a soluble nitrogen source, and accordingly the cellulase is low in cost and convenient to prepare and use. The cellulase prepared by the method is low in cost, high in activity, wide in pH application range, high in stability and short in production cycle.

Description

technical field [0001] The invention belongs to the field of biochemistry, in particular to a preparation method of cellulase. Background technique [0002] Over the years, the biotransformation of cellulose into useful products has been an important research field of biotechnology, but there are very few microorganisms that can directly decompose and use cellulose as a carbon source under natural conditions, especially under neutral conditions. fungi are even rarer. [0003] At present, most of the reports on the research and production of cellulase are about fungi that can decompose cellulosic raw materials under acidic conditions, and there is almost a blank under neutral conditions. The market prospect of neutral cellulase is very good. my country is a big textile country, which needs more than 50,000 tons of cellulase for textile use every year. At present, neutral cellulase is mainly imported, and there is an urgent need to develop a cost-effective and easy-to-operat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12R1/07
Inventor 李秋园代淑梅
Owner ZHONGRONG TECH CORP LTD
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