The Chryseobacterium meningosepticum PafA enzyme: prototype of a new enzyme family of prokaryotic phosphate-irrepressible alkaline phosphatases?
Chryseobacterium meningosepticum is an aerobic Gram-negative rod widely distributed in natural environments. Unlike many bacteria, it produces a phosphate-irrepressible periplasmic alkaline phosphatase (AP). This work describes cloning of the gene encoding that enzyme from C. meningosepticum CCUG 4310 (NCTC 10585), and preliminary characterization of its product. The gene, named pafA, encodes a protein (PafA) of 546 amino acids with a calculated molecular mass of the mature peptide of 58682 Da. PafA exhibits high sequence identity with the PhoV AP of Synechococcus PCC 7942 (49.9% identity) and with the Cda Ca(2+)-dependent ATPase of Myroides odoratus (51.9% identity), while being more distantly related to the PhoD AP of Zymomonas mobilis (22.1% identity) and to the PhoA AP of Escherichia coli (14.0% identity). PafA was partially purified; it exhibits optimal activity at pH 8.5 and is active towards a broad spectrum of substrates including both phosphomonoesters and ATP, with preferential activity for the latter compound. The present findings allow definition of a new family of APs including 60 kDa, periplasmic enzymes whose expression is not influenced by freely available P(i) in the medium. Moreover, PafA can be considered an evolutionary intermediate between Ca(2+)-ATPase of M. odoratus and the APs PhoV of Synechococcus PCC 7942 and PhoD of Z. mobilis.
Berlutti
Francesca
F
Dipartimento di Scienze di Sanità Pubblica, Università di Roma 'La Sapienza', Piazzale A. Moro, 5, 00185 Rome, Italy1.
Passariello
Claudio
C
Dipartimento di Scienze di Sanità Pubblica, Università di Roma 'La Sapienza', Piazzale A. Moro, 5, 00185 Rome, Italy1.
Selan
Laura
L
Dipartimento di Scienze di Sanità Pubblica, Università di Roma 'La Sapienza', Piazzale A. Moro, 5, 00185 Rome, Italy1.
Thaller
Maria Cristina
MC
Dipartimento di Biologia, Università di Roma 'Tor Vergata', 00133 Rome, Italy2.
Rossolini
Gian Maria
GM
Dipartimento di Biologia Molecolare, Sezione di Microbiologia, Università di Siena, 53100 Siena, Italy3.
D000469:Alkaline Phosphatase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D003001:Cloning, Molecular; D004794:Enzyme Repression; D005417:Flavobacterium / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D019897:Periplasm / Q000201:enzymology; D010710:Phosphates / Q000494:pharmacology; D010802:Phylogeny; D017422:Sequence Analysis, DNA
D001426:Bacterial Proteins; D010710:Phosphates; D000469:Alkaline Phosphatase; C449204:periplasmic alkaline phosphatase, Chryseobacterium meningosepticum