Cloning and expression of a neutral phosphatase gene from Treponema denticola.
We have isolated and characterized a neutral phosphatase gene, phoN, from Treponema denticola ATCC 35405. The gene was isolated from a T. denticola clone bank constructed in the medium-copy-number plasmid vector pMCL19. Subcloning and nucleotide sequencing of the DNA insert from one phosphatase clone, pTph14, revealed that the activity corresponded to an open reading frame consisting of 1,027 bp coding for a 37.9-kDa protein. Hydrophobicity analysis indicated that the protein exhibits some hydrophobic regions. Indeed, partial purification of the phosphatase suggested that the enzyme was membrane associated both in T. denticola and in the Escherichia coli clone. The pH optimum of the enzyme, approximately pH 6.4, indicated that it corresponded to a neutral phosphatase activity from T. denticola. An examination of possible natural substrates for the enzyme suggested that this enzyme hydrolyzes nucleoside di- and triphosphates. Northern (RNA) blot analysis revealed that this phosphatase gene is not likely to be present in an operon structure.
Ishihara
K
K
Department of Oral Biology, State University of New York at Buffalo 14214.
Kuramitsu
H K
HK
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics* / Q000378:metabolism; D012329:RNA, Bacterial / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D015183:Restriction Mapping; D013379:Substrate Specificity; D014208:Treponema / Q000201:enzymology / Q000235:genetics*
D004269:DNA, Bacterial; D012329:RNA, Bacterial; D012333:RNA, Messenger; D010744:Phosphoric Monoester Hydrolases