Characterization of arylsulfatase isoenzymes from Pseudomonas aeruginosa

Abstract

A method is described for the separation and isolation of two electrophoretically distinct arylsulfatases (arylsulfatase EC.3.1.6.1) from Pseudomonas aeruginosa.Characterization of the enzymes revealed that they were type I arylsulfatases, with similar kinetic properties. They differed, however, in respect to charge, pH optima for substrate hydrolysis, and activation by anions. In addition, the enzymes displayed dual pH optima curves with both substrates used and the curious property of a shift in pH optima with varied p-nitrophenyl sulfate concentrations.

Riding the sulfur cycle – metabolism of sulfonates and sulfate esters in Gram-negative bacteria

“…Thus, Proteus rettgeri was reported to contain nine arylsulfatase isozymes, with pH optima at 6.7 and 8.3 [105,112], Serratia marcescens contains multiple isozymes with a broad pH optimum of 6.8^8.2 [107], P. aeruginosa has two isozymes with pH optima 9.0 and 8.4 [109,113], and Salmonella typhimurium has ¢ve enzymes, all with optimum pH 6.7 [102]. It should be noted, however, that multiple arylsulfatase bands have also been reported in two species (P. aeruginosa and K. pneumoniae) where later work revealed only one functional arylsulfatase gene.…”

“…For example, if one of the three sulfatases present on the genome of P. aeruginosa were cryptic in one isolate, and expressed in another, this would explain the observed di¡erences in the number of isozymes in di¡erent reports [11,109]. Several species of enteric bacteria, in particular, seem to contain several sulfatase genes.…”

“…Bacterial arylsulfatases have been identi¢ed and studied in a variety of species, including the enterobacteria (Klebsiella [98^100], Salmonella [101,102], Enterobacter [103,104], Proteus [105,106], Serratia [107]), pseudomonads (Pseudomonas [11,108,109], Comamonas [110]), mycobacteria, and cyanobacteria [111]. In recent years, however, genetic and biochemical studies have concentrated primarily on the arylsulfatase enzymes from Pseudomonas aeruginosa and Klebsiella pneumoniae, at least in part because these enzymes are closely related to the human arylsulfatases.…”

“…Two main conclusions can be drawn from the early biochemical studies: (i) most of the strains studied contained multiple arylsulfatase isozymes, and (ii) the arylsulfatase enzymes can be divided into two groups according to pH optimum, with one group showing optimal activity at pH values 6.5^7.1, and the other group at higher pH values of 8.3^9.0. Thus, Proteus rettgeri was reported to contain nine arylsulfatase isozymes, with pH optima at 6.7 and 8.3 [105,112], Serratia marcescens contains multiple isozymes with a broad pH optimum of 6.8^8.2 [107], P. aeruginosa has two isozymes with pH optima 9.0 and 8.4 [109,113], and Salmonella typhimurium has ¢ve enzymes, all with optimum pH 6.7 [102]. It should be noted, however, that multiple arylsulfatase bands have also been reported in two species (P. aeruginosa and K. pneumoniae) where later work revealed only one functional arylsulfatase gene.…”

Purification and properties of arylsulphatase A from rabbit testis

“…Arylsulphatases A and B can be distinguished by their different pH optima, substrate affinities, anomalous kinetics and electrophoretic mobilities (Farooqui & Bachhawat, 1972). Arylsulphatase isoenzymes from bacterial sources can be characterized by their electrophoretic mobilities (Delisle & Milazzo, 1972). Arylsulphatases A and B can be separated by fractional precipitation with acetone (Roy, 1953), by paper electrophoresis (Roy, 1958) and by gel filtration on Sephadex G-200 (Wynn, 1966;.…”