Waddell Grace L, Gilmer Caroline R, Taylor Nicholas G, Reveral John Randolf S, Forconi Marcello, Fox Jennifer L
Department of Chemistry and Biochemistry, College of Charleston, 66 George St., Charleston, SC, USA 29424.
Department of Chemistry and Biochemistry, College of Charleston, 66 George St., Charleston, SC, USA 29424.
Biochem Biophys Res Commun. 2017 Sep 16;491(2):382-387. doi: 10.1016/j.bbrc.2017.07.092. Epub 2017 Jul 15.
The eukaryotic enzyme Bds1 in Saccharomyces cerevisiae is a metallo-β-lactamase-related enzyme evolutionarily originating from bacterial horizontal gene transfer that serves an unknown biological role. Previously, Bds1 was reported to be an alkyl and aryl sulfatase. However, we demonstrate here that Bds1 acts on primary alkyl sulfates (of 6-12 carbon atoms) but not the aryl sulfates p-nitrophenyl sulfate and p-nitrocatechol sulfate. The apparent catalytic rate constant for hydrolysis of the substrate 1-hexyl sulfate by Bds1 is over 100 times lower than that of the reaction catalyzed by its bacterial homolog SdsA1. We show that Bds1 shares a catalytic mechanism with SdsA1 in which the carbon atom of the sulfate ester is the subject of nucleophilic attack, rather than the sulfur atom, resulting in C-O bond lysis. In contrast to SdsA1 and another bacterial homolog with selectivity for secondary alkyl sulfates named Pisa1, Bds1 does not show any substantial activity towards secondary alkyl sulfates. Neither Bds1 nor SdsA1 have any significant activity towards a branched primary alkyl sulfate, primary and secondary steroid sulfates, or phosphate diesters. Therefore, the enzymes homologous to SdsA1 that have been identified and characterized thus far vary in their selectivity towards primary and secondary alkyl sulfates but do not exhibit aryl sulfatase activity.
酿酒酵母中的真核酶Bds1是一种与金属β-内酰胺酶相关的酶,它在进化上起源于细菌水平基因转移,其生物学功能未知。此前,有报道称Bds1是一种烷基和芳基硫酸酯酶。然而,我们在此证明,Bds1作用于(含有6至12个碳原子的)伯烷基硫酸盐,但不作用于芳基硫酸盐对硝基苯硫酸盐和对硝基邻苯二酚硫酸盐。Bds1催化底物1-己基硫酸盐水解的表观催化速率常数比其细菌同源物SdsA1催化的反应低100多倍。我们表明,Bds1与SdsA1具有相同的催化机制,即硫酸酯的碳原子是亲核攻击的对象,而不是硫原子,从而导致C-O键裂解。与SdsA1和另一种对仲烷基硫酸盐具有选择性的细菌同源物Pisa1不同,Bds1对仲烷基硫酸盐没有任何显著活性。Bds1和SdsA1对支链伯烷基硫酸盐、伯和仲类固醇硫酸盐或磷酸二酯均无任何显著活性。因此,迄今为止已鉴定和表征的与SdsA1同源的酶对伯烷基硫酸盐和仲烷基硫酸盐的选择性各不相同,但均不表现出芳基硫酸酯酶活性。