Cloning, sequence and characterization of the polyphosphate kinase-encoding gene (ppk) of Klebsiella aerogenes.
Polyphosphate kinase (PPK) catalyzes the formation of polyphosphate (polyP). The PPK-encoding gene (ppk) has been cloned from Klebsiella aerogenes ATCC9621. The gene possessed an open reading frame of 2055 bp capable of encoding a putative polypeptide with a deduced M(r) of 80,157. This polypeptide showed 93% similarity to the Escherichia coli PPK. The nucleotide sequence of the promoter region of K. aerogenes ppk differed from that of the previously sequenced E. coli ppk. A putative pho box sequence was found in the promoter region of K. aerogenes ppk. The expression of lacZ from the ppk promoter was increased in E. coli MV1184 under conditions of phosphate (Pi) limitation, but not in E. coli ANCS3 (phoB-), indicating that the ppk promoter is regulated by the phoB product. Increased levels of specific PPK activity were shown by expressing the cloned ppk at high levels, resulting in increased accumulation of polyP in E. coli.
Kato
J
J
Department of Fermentation Technology, Hiroshima University, Japan.
Yamamoto
T
T
Yamada
K
K
Ohtake
H
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D005798:Genes, Bacterial; D007709:Klebsiella / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D017856:Phosphotransferases (Phosphate Group Acceptor) / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping
D004269:DNA, Bacterial; D017856:Phosphotransferases (Phosphate Group Acceptor); C022424:polyphosphate kinase