Characterization of endoglucanase cela from the rumen bacterium Clostridium longisporum

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dc.contributor.advisorThomson, Jennifer Annen_ZA
dc.contributor.authorMittendorf, Volkeren_ZA
dc.date.accessioned2016-08-24T12:54:06Z
dc.date.available2016-08-24T12:54:06Z
dc.date.issued1994en_ZA
dc.descriptionBibliography: pages 84-101.en_ZA
dc.description.abstractCellulose is the most abundant organic compound on earth and offers great potential as a source of renewable energy and other chemicals. Cellulases are being studied to elucidate the enzymatic degradation of cellulose. We are interested in the molecular mechanisms of microbial degradation of cellulose in the rumen. The long-term aim is the potential genetic modification of lignocellulolytic activities in the rumen. · Clostridium longisporum was obtained from Varel (1989) because oxygen-resistant endospores might be suitable "vectors" for the introduction of genetically modified enzyme systems into the rumen via animal feeds. It is a sporadically occurring rumen bacterium and its role in ruminal cellulolysis is unclear. The aim of this project was the initial characterization of cellulases produced by C. longisporum. The celA gene was obtained by screening a library of C. longisporum genomic DNA in Escherichia colifor clones expressing CMCase activity. Approximately 38 CMCase-positive clones were obtained and the plasmid pCM4 was isolated from the clone expressing the highest activity. Southern analysis indicated that another plasmid, pCM64, contained a larger insert including the insert of pCM4. A total of 3620 bp were sequenced and a 1548-bp open reading frame, termed celA, was found. This gene showed homology with other endo-B-1,4-glucanases from family 5 (Henrissat & Bairoch, 1993). Plasmid pCM64 was found to contain the whole celA gene encoding endoglucanase CelA, while pCM4 has a 5'-truncated gene, termed celM5', which encodes a fusion protein, CelMN', that was initiated from an ATG codon in the vector. Sequence analysis of celA revealed the presence of a type I cellulose-binding domain (Beguin & Aubert, 1994) at the COOH-terminus of CelA.en_ZA
dc.identifier.apacitationMittendorf, V. (1994). <i>Characterization of endoglucanase cela from the rumen bacterium Clostridium longisporum</i>. (Thesis). University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology. Retrieved from http://hdl.handle.net/11427/21496en_ZA
dc.identifier.chicagocitationMittendorf, Volker. <i>"Characterization of endoglucanase cela from the rumen bacterium Clostridium longisporum."</i> Thesis., University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology, 1994. http://hdl.handle.net/11427/21496en_ZA
dc.identifier.citationMittendorf, V. 1994. Characterization of endoglucanase cela from the rumen bacterium Clostridium longisporum. University of Cape Town.en_ZA
dc.identifier.ris TY - Thesis / Dissertation AU - Mittendorf, Volker AB - Cellulose is the most abundant organic compound on earth and offers great potential as a source of renewable energy and other chemicals. Cellulases are being studied to elucidate the enzymatic degradation of cellulose. We are interested in the molecular mechanisms of microbial degradation of cellulose in the rumen. The long-term aim is the potential genetic modification of lignocellulolytic activities in the rumen. · Clostridium longisporum was obtained from Varel (1989) because oxygen-resistant endospores might be suitable "vectors" for the introduction of genetically modified enzyme systems into the rumen via animal feeds. It is a sporadically occurring rumen bacterium and its role in ruminal cellulolysis is unclear. The aim of this project was the initial characterization of cellulases produced by C. longisporum. The celA gene was obtained by screening a library of C. longisporum genomic DNA in Escherichia colifor clones expressing CMCase activity. Approximately 38 CMCase-positive clones were obtained and the plasmid pCM4 was isolated from the clone expressing the highest activity. Southern analysis indicated that another plasmid, pCM64, contained a larger insert including the insert of pCM4. A total of 3620 bp were sequenced and a 1548-bp open reading frame, termed celA, was found. This gene showed homology with other endo-B-1,4-glucanases from family 5 (Henrissat & Bairoch, 1993). Plasmid pCM64 was found to contain the whole celA gene encoding endoglucanase CelA, while pCM4 has a 5'-truncated gene, termed celM5', which encodes a fusion protein, CelMN', that was initiated from an ATG codon in the vector. Sequence analysis of celA revealed the presence of a type I cellulose-binding domain (Beguin & Aubert, 1994) at the COOH-terminus of CelA. DA - 1994 DB - OpenUCT DP - University of Cape Town LK - https://open.uct.ac.za PB - University of Cape Town PY - 1994 T1 - Characterization of endoglucanase cela from the rumen bacterium Clostridium longisporum TI - Characterization of endoglucanase cela from the rumen bacterium Clostridium longisporum UR - http://hdl.handle.net/11427/21496 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/21496
dc.identifier.vancouvercitationMittendorf V. Characterization of endoglucanase cela from the rumen bacterium Clostridium longisporum. [Thesis]. University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology, 1994 [cited yyyy month dd]. Available from: http://hdl.handle.net/11427/21496en_ZA
dc.language.isoengen_ZA
dc.publisher.departmentDepartment of Molecular and Cell Biologyen_ZA
dc.publisher.facultyFaculty of Scienceen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.subject.otherMolecular and Cell Biologyen_ZA
dc.titleCharacterization of endoglucanase cela from the rumen bacterium Clostridium longisporumen_ZA
dc.typeDoctoral Thesis
dc.type.qualificationlevelDoctoral
dc.type.qualificationnamePhDen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceThesisen_ZA
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