Molecular characterization of polyphosphate kinase (ppk) gene from Serratia marcescens.
To understand the mechanism of phosphate accumulation, a gene encoding polyphosphate kinase (PPK) was cloned from the genomic library of Serratia marcescens by Southern hybridization. From the nucleotide sequence of a 4 kb DNA fragment, an open reading frame of 2063 nucleotides was identified encoding a protein of 686 amino acids with molecular mass of 70 kDa. The potential CRP binding site and pho box sequence were found upstream of the putative promoter in the regulatory region. The expression of PPK resulted in the formation of inclusion bodies and the product was active at low temperature. The E. coli strain harboring plasmid pSPK5 with ppk gene increased enzyme activity of polyphosphate kinase, resulting in increased accumulation of polyphosphate in E. coli.
Lee
Seung-Jin
SJ
Division of Biotechnology, Faculty of Natural Resources and Life Science, Dong-A University, Busan 604-714, Korea.
Song
Ok-Ryul
OR
Lee
Young-Choon
YC
Choi
Yong-Lark
YL
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004789:Enzyme Activation; D004926:Escherichia coli / Q000737:chemistry / Q000201:enzymology / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D008969:Molecular Sequence Data; D015394:Molecular Structure; D017856:Phosphotransferases (Phosphate Group Acceptor) / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D012706:Serratia marcescens / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development
D017856:Phosphotransferases (Phosphate Group Acceptor); C022424:polyphosphate kinase