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| Isolation of Thermophilic Cellulose-Decomposing Bacteria and Cloning and Expression of an Endoglucanase Gene |
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| KeyWord:thermophilic bacteria; endoglucanase gene; cloning; expression; domain |
| Author Name | Affiliation | | TIAN Wei | Nanjing Institute of Environmental Science of the Ministry of Environmental Protection of China, Nanjing 210042, China | | WANG Lei | Nanjing Institute of Environmental Science of the Ministry of Environmental Protection of China, Nanjing 210042, China | | LI Gang | Nanjing Institute of Environmental Science of the Ministry of Environmental Protection of China, Nanjing 210042, China | | WANG Zhen | Nanjing Institute of Environmental Science of the Ministry of Environmental Protection of China, Nanjing 210042, China | | LI Yan | Nanjing Institute of Environmental Science of the Ministry of Environmental Protection of China, Nanjing 210042, China | | ZHANG Ji-bing | Nanjing Institute of Environmental Science of the Ministry of Environmental Protection of China, Nanjing 210042, China |
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| Abstract: |
| Thermophilic cellulose-decomposing bacteria could shorten digesting time of composts. Here we isolated eight strains of thermopilic cellulose-decomposing bacteria from composts produced from dairy manure and rice chaff using filter paper as a sole carbon source. After 48 hour cultivation on CMC-congo red medium, strains T1 and T2 had bigger clear zones than the other strains. Based on the morphological, physiological and biochemical properties as well as 16S rRNA homology analysis, the strains T1 and T2 were identified as Bacillus cereus and Bacillus subtilis respectively. Endoglucanase gene of T2 was cloned and domains analysis showed that the endoglucanase gene contained an open reading frame of 1500 bp, corresponding to 499 amino acids. The endoglucanase was comprised of two discontinuous domains：one was N terminal catalytic domain belonging to glycosyl hydrolase family 5, and the other was C terminal substrate binding domain belonging to carbohydrate binding domain family 3. Using PET28a as the expression plasmid, the endoglucanase gene of strain T2 was successfully expressed in E. coli BL21（DE3）. SDS-PAGE analysis showed that molecular weight of the expressed protein was about 50 KD and the concentration was about 93.14 mg·L-1. |
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