Find in Library

Arylsulfatases exhibit great potential in industry for desulfation applications, but less is known about the metallo-β-lactamase (MBL) fold arylsulfatases. To learn more about them, an MBL fold arylsulfatase from <i>Pseudoalteromonas atlantica</i> T6c (PaAst) was identified and characterized, and its structure was elaborated in this study. PaAst was sequence analyzed, heterologously expressed in <i>E. coli</i>, purified by Ni²⁺-NTA resin affinity chromatography and size-exclusion chromatography, functionally studied by <i>p</i>-nitrophenyl sulfate (<i>p</i>NPS), and crystallized for structure determination. The MBL fold arylsulfatase was identified by sequence analysis and confirmed by enzymatic assay on <i>p</i>NPS with K_m 1.00 mM and V_max 60.80 U/mg at 50 °C and pH 7.5. Furthermore, its crystals were obtained in 0.2 M sodium thiocyamate, 20% PEG3350, and its structure was determined at 2.0 Å that formed a dimer with MBL fold. Our work highlighted the MBL fold arylsulfatases from structural insights and could be the theoretical foundation for investigations into their catalytic mechanism.