Molecular cloning of a phytase gene (phy M) from Pseudomonas syringae MOK1.
A phytase gene (phy M) was cloned from Pseudomonas syringae MOK1 by two steps of degenerate PCR and inverse PCR. This gene consists of 1,287 nucleotides and encodes a polypeptide of 428 amino acids with a deduced molecular mass of 46,652 kDa. Based on its amino acid sequence, the Phy M shares the active site RHGXRXP and HD sequence motifs, typically characterized by histidine acid phosphatases familly. Each phy M gene fragment encoding mature Phy M with its own signal sequence (pEPSS) and without (pEPSM) was subcloned into the E. coli BL21 (DE3) expression vector, pET22b (+). The enzyme activity in crude extracts of clone pEPSM was 2.514 Umg(-1) of protein, and about 10-fold higher than that of clone pEPSS.
Cho
Jaiesoon
J
Inositide Signaling Group, National Institute of Environmental Health Sciences, NIH, DHSS, Research Triangle Park, PO Box 12233, NC 27709, USA. cho3@niehs.nih.gov
Lee
Changwhan
C
Kang
Seungha
S
Lee
Jaecheon
J
Lee
Honggu
H
Bok
Jinduck
J
Woo
Junghee
J
Moon
Yangsoo
Y
Choi
Yunjaie
Y
D010832:6-Phytase / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D044224:Pseudomonas syringae / Q000201:enzymology / Q000235:genetics*
D010832:6-Phytase