Purification, Characterization, and Structural Studies of a Sulfatase from Pedobacter yulinensis

• Published in: Molecules (Basel, Switzerland) Vol. 27; no. 1; p. 87
• Main Authors: Schlachter, Caleb R, O'Malley, Andrea, Grimes, Linda L, Tomashek, John J, Chruszcz, Maksymilian, Lee, L Andrew
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 24.12.2021; MDPI
• Subjects: 17β-Estradiol; 4-methylumbelliferyl sulfate; Amino Acid Sequence; Amino acids; Binding Sites; Biotechnology; Carbohydrates; Catalytic Domain; Chemical Fractionation - methods; Chromatography; Cortisol; Crystal structure; crystallography; Drug abuse; Enzyme Stability; Enzymes; Flavones; formylglycine; Humans; hydrolase; Mass spectrometry; Metabolism; Metabolites; Models, Molecular; Molecular Structure; Pedobacter - enzymology; Pedobacter yulinensis; Peptides; Phosphates; Post-translation; Protein Conformation; Protein Multimerization; Proteins; Recombinant Proteins; Scientific imaging; Sex hormones; Steroid hormones; Steroids; Structural analysis; Structure-Activity Relationship; Substrate Specificity; Substrates; sulfatase; Sulfatases - chemistry; Sulfatases - isolation & purification; Sulfatases - metabolism; Sulfates; Tuberculosis; crystallography; Pedobacter yulinensis; hydrolase; sulfatase; arylsulfatase; 4-methylumbelliferyl sulfate; formylglycine
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules27010087

Sulfatases are ubiquitous enzymes that hydrolyze sulfate from sulfated organic substrates such as carbohydrates, steroids, and flavones. These enzymes can be exploited in the field of biotechnology to analyze sulfated metabolites in humans, such as steroids and drugs of abuse. Because genomic data far outstrip biochemical characterization, the analysis of sulfatases from published sequences can lead to the discovery of new and unique activities advantageous for biotechnological applications. We expressed and characterized a putative sulfatase (PyuS) from the bacterium . PyuS contains the (C/S)XPXR sulfatase motif, where the Cys or Ser is post-translationally converted into a formylglycine residue (FGly). His-tagged PyuS was co-expressed in with a formylglycine-generating enzyme (FGE) from and purified. We obtained several crystal structures of PyuS, and the FGly modification was detected at the active site. The enzyme has sulfatase activity on aromatic sulfated substrates as well as phosphatase activity on some aromatic phosphates; however, PyuS did not have detectable activity on 17α-estradiol sulfate, cortisol 21-sulfate, or boldenone sulfate.