Cloning and expression of aClostridium thermocellum dna fragment that encodes a protein related to cellulosome component sL
| Autor: | M.P.M. Romaniec, U. T. Gerngross, T. Kobayashi, Arnold L. Demain, U. Fauth |
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| Rok vydání: | 1991 |
| Předmět: |
Glycoside Hydrolases
Protein subunit Restriction Mapping Gene Expression Bioengineering macromolecular substances Molecular cloning Biology Applied Microbiology and Biotechnology Biochemistry Substrate Specificity law.invention Cellulosome chemistry.chemical_compound Restriction map Cellulase law Protein purification Cloning Molecular Cellulose Molecular Biology Clostridium technology industry and agriculture Membrane Proteins General Medicine biology.organism_classification Molecular biology Recombinant Proteins chemistry Recombinant DNA Clostridium thermocellum DNA Biotechnology |
| Zdroj: | Applied Biochemistry and Biotechnology. 31:119-134 |
| ISSN: | 1559-0291 0273-2289 |
| Popis: | Antibodies raised against the SL subunit of the Clostridium thermocellum cellulosome were used to screen a library of C. thermocellum chromosomal DNA fragments constructed in the vector lambda gt11. A DNA fragment that encoded a polypeptide that crossreacted with the anti-SL antibodies was isolated and its restriction map elucidated. No similarity with other previously cloned DNA fragments has been found. The anti-SL crossreacting polypeptide was isolated from recombinant Escherichia coli and found to have a mol mass of 37,000 Da and to possess low levels of CMCase and Avicelase activity. Using CMC as the substrate, a temperature optimum of 55 degrees C and a pH optimum of 6.6 were observed. These properties were compared to those of C. thermocellum SL isolated by electroelution from an SDS gel, which was also found to possess low levels of CMCase and Avicelase activities. In addition, the SL proteins produced in C. thermocellum and E. coli were able to interact positively against Avicel with an endoglucanase (Ss) purified from the C. thermocellum crude cellulase preparation, and with a recombinant protein that crossreacted with anti-Ss antibodies. |
| Databáze: | OpenAIRE |
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