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Cloning and expression of a pectate lyase from the oral spirochete Treponema pectinovorum ATCC 33768
Article first published online: 9 JAN 2006
DOI: 10.1016/S0378-1097(03)00639-6
1574-6968/asset/cover.gif?v=1&s=069bb2c224e243333d4486580fbd90d9ebeffa6b "FEMS Microbiology Letters")
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How to Cite
Walker, S. G. and Ryan, M. E. (2003), Cloning and expression of a pectate lyase from the oral spirochete Treponema pectinovorum ATCC 33768. FEMS Microbiology Letters, 226: 385–390. doi: 10.1016/S0378-1097(03)00639-6
Publication History
- Issue published online: 9 JAN 2006
- Article first published online: 9 JAN 2006
- Received 7 June 2003, Revised 5 August 2003, Accepted 11 August 2003
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Keywords:
- Pectate lyase;
- Pectinase;
- Oral treponeme;
- Spirochete;
- Periodontal disease;
- Treponema pectinovorum
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.