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Cellulase producing bacteria and application thereof

A cellulase and bacteria-producing technology, applied in the field of biochemistry, can solve the problems of high animal feeding costs, environmental pollution, incomplete degradation of cellulose components, etc., and achieve shortened production cycle, broad application prospects, and pH applicable range. wide effect

Active Publication Date: 2015-11-11
INST OF AQUATIC LIFE ACAD SINICA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in modern aquaculture intensive farming, there are widespread problems of incomplete degradation of a large number of cellulose components. On the one hand, these problems have caused high animal feeding costs, and on the other hand, they have brought a lot of environmental pollution. Therefore, a suitable aquatic Strains and systems that efficiently express neutral cellulase in a neutral environment have great potential significance for the aquatic industry

Method used

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  • Cellulase producing bacteria and application thereof
  • Cellulase producing bacteria and application thereof
  • Cellulase producing bacteria and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Isolation, screening and identification of Bacillus megaterium SMY0001

[0023] Primary screening of bacterial strains: Collect 6 parts of the hindgut content of healthy grass carp, weigh 0.5g samples respectively, add 4.5mL sterile water to prepare a suspension, kill non-spore vegetative bodies in a water bath at 80°C, and then spread on carboxyl carp On the sodium methylcellulose screening plate, cultivate at 37°C for 48 hours, and obtain 18 strains of bacteria.

Embodiment 218

[0024] The identification of embodiment 218 cellulolytic bacteria:

[0025] After collecting the culture fluid of 18 cellulolytic bacteria, the genomic DNA was extracted and used as a template for PCR to amplify the full-length 16S of the bacteria. After the PCR products were recovered and cloned, they were sequenced, and the sequencing results were recorded in the GenBank database using BLAST (BasicLocalAlignmentSearchTool) software. Perform a sequence similarity search to determine the strain information. The results showed that 12 of the 18 bacterial strains were Bacillus.

Embodiment 3

[0026] Embodiment 3, screening the bacterial classification SMY0001 with the highest enzymatic activity:

[0027] Preparation of crude enzyme liquid: Inoculate 12 strains of bacillus into carboxymethylcellulose medium (composition: 1% sodium carboxymethylcellulose, 1% yeast extract, 1.5% inorganic salt, 96.5% water, pH7 .0), cultured at 55°C for 7 days, and centrifuged at 8000r / min for 20min to obtain crude enzyme solution.

[0028] Determination of CMC enzyme activity: use 1% sodium carboxymethylcellulose (CMC) as the substrate, add enzyme solution with appropriate dilution factor, react at 50°C, 80r / min, pH4-10 for 30 minutes, and measure the enzyme produced by enzymolysis Glucose volume. A unit of CMC enzyme activity is defined as: the amount of enzyme that produces one milligram of glucose per milliliter of enzyme solution per hour under standard reaction conditions, expressed in u / ml. From the test results, the Bacillus megaterium strain SMY0001, which highly expresses c...

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Abstract

The invention discloses a cellulase producing bacteria classified and named bacillus megatherium. The bacteria strain is preserved in China Center for Type Culture Collection (CCTCC) at May 21, 2015 and is assigned the accession number of CCTCC M2015320. The bacteria strain is used for preparing high-activity cellulase under a neutral pH and is significantly better than an acidic cellulase in the field of water-washing enzyme application. In the invention, sodium carboxymethylcellulose is employed as a carbon source, which is excellent than a fiber raw material in the aspects of transportation, material feeding and disinfection and the like. With urea as a soluble nitrogen source, the cellulase producing bacteria is low in cost and is convenient to prepare and use. According to the invention, the cellulase is high in activity, is wide in application range and good in stability to pH, and is short in production period.

Description

technical field [0001] The invention belongs to the field of biochemistry, and in particular relates to a cellulase-producing bacterium and application thereof. Background technique [0002] Cellulase is a general term for multi-component enzymes that degrade cellulose to generate glucose. It has huge market potential in the fields of feed, alcohol, textile and food. The source of cellulase is very wide, insects, molluscs, protozoa, bacteria , actinomycetes and fungi can produce cellulase. For a long time, people have studied the acid cellulase produced by fungi the most in-depth and extensively. However, with the expansion of the application field of cellulase, the traditional acid cellulase produced by fungi cannot be applied to all industries, while the acid cellulase produced by bacteria Neutral and alkaline cellulase has increasingly shown its unique advantages in industries such as textiles, detergents, papermaking, and environmental pollution treatment. However, the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/42A23K1/18A23K1/165C12R1/11
Inventor 王桂堂熊凡吴山功邹红李文祥
Owner INST OF AQUATIC LIFE ACAD SINICA
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