Expression of Treponema denticola oligopeptidase B in Escherichia coli.
Treponema denticola is a small anaerobic spirochete often isolated from periodontal lesions and closely associated with periodontal diseases. This bacterium possesses a particular arginine peptidase activity (previously called "BANA-peptidase" or "trypsin-like enzyme") that is common to the three cultivable bacterial species most highly associated with severe periodontal disease. We recently reported the identification of the opdB locus that encodes the BANA-peptidase activity of T. denticola through DNA sequencing and mutagenesis studies. In the present study, we report expression of T. denticola OpdB peptidase in Escherichia coli. The opdB PCR product was cloned into pET30b and then transformed into the E. coli BL21 (DE3)/pLysS expression strain. Assays of enzymatic activities in E. coli containing T. denticola opdB showed BANA-peptidase activity similar to that of T. denticola. Availability of this recombinant expression system producing active peptidase will facilitate characterization of the potential role of this peptidase in periodontal disease etiology.
Lee
Si Young
SY
Department of Oral Microbiology, College of Dentistry, Research Institute of Oral Science, Kangnung National University, Kangnung, Korea.
Fenno
J Christopher
JC
D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics*; D001586:Benzoylarginine Nitroanilide / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D012697:Serine Endopeptidases / Q000096:biosynthesis* / Q000235:genetics*; D014169:Transformation, Bacterial; D014208:Treponema / Q000201:enzymology* / Q000235:genetics*
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D001586:Benzoylarginine Nitroanilide; D012697:Serine Endopeptidases; C090703:oligopeptidase B