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Characterisation of two bifunctional cellulase-xylanase enzymes isolated from a bovine rumen metagenome library

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dc.contributor.author Rashamuse, KJ
dc.contributor.author Visser, Daniel F
dc.contributor.author Hennessy, F
dc.contributor.author Kemp, J
dc.contributor.author Van der Merwe, MP
dc.contributor.author Badenhorst, J
dc.contributor.author Ronneburg, T
dc.contributor.author Francis-Pope, R
dc.contributor.author Brady, D
dc.date.accessioned 2013-03-25T06:19:26Z
dc.date.available 2013-03-25T06:19:26Z
dc.date.issued 2013-02
dc.identifier.citation Rashamuse, KJ, Visser, DF, Hennessy, F, Kemp, J, Van der Merwe, MP, Badenhorst, J, Ronneburg, T, Francis-Pope, R and Brady D. 2013. Characterisation of two bifunctional cellulase-xylanase enzymes isolated from a bovine rumen metagenome library. Current Microbiology, vol. 66(2), pp 145-151 en_US
dc.identifier.issn 145-151
dc.identifier.uri http://www.ncbi.nlm.nih.gov/pubmed/23086538
dc.identifier.uri http://hdl.handle.net/10204/6586
dc.description Copyright: 2013 Springer Verlag. This is the pre/post print version of the work. The definitive version is published in Current Microbiology, vol. 66(2), pp 145-151 en_US
dc.description.abstract Ruminant digestive tract microbes hydrolyse plant biomass, and the application of metagenomic techniques can provide good coverage of their glycosyl hydrolase enzymes. A metagenomic library of circa 70,000 fosmids was constructed from bacterial DNA isolated from bovine rumen and subsequently screened for cellulose hydrolysing activities on a CMC agar medium. Two clones were selected based on large clearance zones on the CMC agar plates. Following nucleotide sequencing, translational analysis and homology searches, two cellulase encoding genes (cel5A and cel5B) belonging to the glycosyl hydrolyse family 5 were identified. Both genes encoded pre-proteins of about 62 kDa, containing signal leader peptides which could be cleaved to form mature proteins of about 60 kDa. Biochemical characterisation revealed that both enzymes showed alkaline pH optima of 9.0 and the temperature optima of 65 °C. Substrate specificity profiling of the two enzymes using 1,4-ß-D-cello- and xylo-oligosaccharides revealed preference for longer oligosaccharides (n = 3) for both enzymes, suggesting that they are endo-cellulases/xylanases. The bifunctional properties of the two identified enzymes render them potentially useful in degrading the ß-1,4 bonds of both the cellulose and hemicellulose polymers. en_US
dc.language.iso en en_US
dc.publisher Springer Verlag en_US
dc.relation.ispartofseries Workflow;10285
dc.subject Enzymes en_US
dc.subject Cellulose polymers en_US
dc.subject Hemicellulose ploymers en_US
dc.title Characterisation of two bifunctional cellulase-xylanase enzymes isolated from a bovine rumen metagenome library en_US
dc.type Article en_US
dc.identifier.apacitation Rashamuse, K., Visser, D. F., Hennessy, F., Kemp, J., Van der Merwe, M., Badenhorst, J., ... Brady, D. (2013). Characterisation of two bifunctional cellulase-xylanase enzymes isolated from a bovine rumen metagenome library. http://hdl.handle.net/10204/6586 en_ZA
dc.identifier.chicagocitation Rashamuse, KJ, Daniel F Visser, F Hennessy, J Kemp, MP Van der Merwe, J Badenhorst, T Ronneburg, R Francis-Pope, and D Brady "Characterisation of two bifunctional cellulase-xylanase enzymes isolated from a bovine rumen metagenome library." (2013) http://hdl.handle.net/10204/6586 en_ZA
dc.identifier.vancouvercitation Rashamuse K, Visser DF, Hennessy F, Kemp J, Van der Merwe M, Badenhorst J, et al. Characterisation of two bifunctional cellulase-xylanase enzymes isolated from a bovine rumen metagenome library. 2013; http://hdl.handle.net/10204/6586. en_ZA
dc.identifier.ris TY - Article AU - Rashamuse, KJ AU - Visser, Daniel F AU - Hennessy, F AU - Kemp, J AU - Van der Merwe, MP AU - Badenhorst, J AU - Ronneburg, T AU - Francis-Pope, R AU - Brady, D AB - Ruminant digestive tract microbes hydrolyse plant biomass, and the application of metagenomic techniques can provide good coverage of their glycosyl hydrolase enzymes. A metagenomic library of circa 70,000 fosmids was constructed from bacterial DNA isolated from bovine rumen and subsequently screened for cellulose hydrolysing activities on a CMC agar medium. Two clones were selected based on large clearance zones on the CMC agar plates. Following nucleotide sequencing, translational analysis and homology searches, two cellulase encoding genes (cel5A and cel5B) belonging to the glycosyl hydrolyse family 5 were identified. Both genes encoded pre-proteins of about 62 kDa, containing signal leader peptides which could be cleaved to form mature proteins of about 60 kDa. Biochemical characterisation revealed that both enzymes showed alkaline pH optima of 9.0 and the temperature optima of 65 °C. Substrate specificity profiling of the two enzymes using 1,4-ß-D-cello- and xylo-oligosaccharides revealed preference for longer oligosaccharides (n = 3) for both enzymes, suggesting that they are endo-cellulases/xylanases. The bifunctional properties of the two identified enzymes render them potentially useful in degrading the ß-1,4 bonds of both the cellulose and hemicellulose polymers. DA - 2013-02 DB - ResearchSpace DP - CSIR KW - Enzymes KW - Cellulose polymers KW - Hemicellulose ploymers LK - https://researchspace.csir.co.za PY - 2013 SM - 145-151 T1 - Characterisation of two bifunctional cellulase-xylanase enzymes isolated from a bovine rumen metagenome library TI - Characterisation of two bifunctional cellulase-xylanase enzymes isolated from a bovine rumen metagenome library UR - http://hdl.handle.net/10204/6586 ER - en_ZA


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