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Volume 147, Issue 10

The PafA enzyme: prototype of a new enzyme family of prokaryotic phosphate-irrepressible alkaline phosphatases?

The GenBank accession number for reported in this paper is AF157621.

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Abstract

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 CCUG 4310 (NCTC 10585), and preliminary characterization of its product. The gene, named , 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 PCC 7942 (499% identity) and with the Cda Ca-dependent ATPase of (519% identity), while being more distantly related to the PhoD AP of (221% identity) and to the PhoA AP of (140% identity). PafA was partially purified; it exhibits optimal activity at pH 85 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 in the medium. Moreover, PafA can be considered an evolutionary intermediate between Ca-ATPase of and the APs PhoV of PCC 7942 and PhoD of .

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Keyword(s): AP, alkaline phosphatase , BCIP, 5-bromo-4-chloro-3-indolyl phosphate , Ca2+-dependent ATPase , Cda , flavobacteria , p-nitrophenyl phosphate , PDP-MG, phenolphthalein diphosphate-methyl green medium , PhoD , PhoV , pNP, p-nitrophenol , pNPP , TBU, Terrific broth supplemented with 1% glucose and TBUXP, TBU agar containing 50 μg BCIP ml−1
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2001-10-01
2025-04-27
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/content/journal/micro/10.1099/00221287-147-10-2831
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The Chryseobacterium meningosepticum PafA enzyme: prototype of a new enzyme family of prokaryotic phosphate-irrepressible alkaline phosphatases?The GenBank accession number for pafA reported in this paper is AF157621.
Microbiology 147, 2831 (2001); https://doi.org/10.1099/00221287-147-10-2831
/content/journal/micro/10.1099/00221287-147-10-2831
/content/journal/micro/10.1099/00221287-147-10-2831
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