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Molecular cloning and expression of a Micromonospora chalcae beta-glucosidase encoding gene in Escherichia coli.

Biomedical Sciences Research Institute Computer Science Research Institute Environmental Sciences Research Institute Nanotechnology & Advanced Materials Research Institute

Winters, A, Gallagher, J, Barron, N, Rollan, A and McHale, AP (1996) Molecular cloning and expression of a Micromonospora chalcae beta-glucosidase encoding gene in Escherichia coli. BIOTECHNOLOGY LETTERS, 18 (12). pp. 1387-1390. [Journal article]

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DOI: 10.1007/BF00129340

Abstract

A Sau3A I genomic library from the actinomycete Micromonospora chalae was constructed in Escherichia coli using the expression vector pUC 18. Using the chromogenic substrate 5-bromo-4-chloro-3-indolyl-beta-glucoside (X-glu), a number of positive recombinant colonies were identified. One of those exhibiting the strongest phenotype contained a recombinant plasmid, pANNA1 which harboured a 4.2kb DNA insert. Using restriction endonuclease site mapping and subcloning strategies a 2.3kb DNA fragment encoding the beta-glucosidase activity was identified. Characterization of the strongly expressed recombinant enzyme demonstrated that it had a dramatically increased thermal stability at 50 degrees C. The Km values obtained for the recombinant enzyme and that from M. chalcae using the substrate beta-nitrophenyl-beta-D-glucoside were 0.19mM and 0.25mM, respectively.

Item Type:Journal article
Faculties and Schools:Faculty of Life and Health Sciences
Faculty of Life and Health Sciences > School of Pharmacy and Pharmaceutical Science
Research Institutes and Groups:Biomedical Sciences Research Institute
Biomedical Sciences Research Institute > Pharmaceutical Science and Practice
ID Code:21526
Deposited By:Professor Anthony McHale
Deposited On:28 Mar 2012 16:21
Last Modified:04 Dec 2012 11:52

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