Cloning and expression of a pectate lyase from the oral spirochete Treponema pectinovorum ATCC 33768 |
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Authors: | Walker Stephen G Ryan Maria E |
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Institution: | Department of Oral Biology and Pathology, School of Dental Medicine, State University of New York, Stony Brook, NY 11794-8702, USA. stephen.walker@stonybrook.edu |
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Abstract: | The pelA gene, encoding a pectate lyase, from Treponema pectinovorum ATCC 33768 was isolated by heterologous expression of a cosmid library in Escherichia coli. In vitro transposon mutagenesis identified an open reading frame of 1293 bp capable of encoding a protein of 430 amino acids with a predicted amino-terminal signal sequence of 21 amino acids. Analysis of the amino acid sequence suggested that it is a member of the polysaccharide lyase family 10 of which all characterized members show pectate lyase activity. An amino-terminal His-tagged recombinant form of PelA was expressed and purified from E. coli. The recombinant enzyme has characteristics common to other bacterial pectate lyases such as an alkaline pH optimum, dependence on calcium ions for activity, and inhibition by zinc ions. |
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Keywords: | Pectate lyase Pectinase Oral treponeme Spirochete Periodontal disease Treponema pectinovorum |
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