Purification and characterization of Mycobacterium tuberculosis 1D-myo-inosityl-2-acetamido-2-deoxy-alpha-D-glucopyranoside deacetylase, MshB, a mycothiol biosynthetic enzyme.

Abstract	Mycothiol (MSH, AcCys-GlcN-Ins) is the major low molecular weight thiol in actinomycetes and is essential for growth of Mycobacterium tuberculosis. MshB, the GlcNAc-Ins deacetylase, is a key enzyme in MSH biosynthesis. MshB from M. tuberculosis was cloned, expressed, purified, and its properties characterized. Values of k(cat) and K(m) for MshB were determined for the biological substrate, GlcNAc-Ins, and several other good substrates. The substrate specificity of MshB was compared to that of M. tuberculosis mycothiol S-conjugate amidase (Mca), a homologous enzyme having weak GlcNAc-Ins deacetylase activity. Both enzymes are metalloamidases with overlapping amidase activity toward mycothiol S-conjugates (AcCySR-GlcN-Ins). The Ins residue and hydrophobic R groups enhance the activity with both MshB and Mca, but changes in the acyl group attached to GlcN have opposite effects on the two enzymes.
Authors	Gerald L Newton, Mary Ko, Philong Ta, Yossef Av-Gay, Robert C Fahey
Journal	Protein expression and purification (Protein Expr Purif) Vol. 47 Issue 2 Pg. 542-50 (Jun 2006) ISSN: 1046-5928 [Print] United States
PMID	16630724 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
Chemical References	Bacterial Proteins Disaccharides Glycopeptides Metalloproteins Pyrazoles Recombinant Proteins Sulfhydryl Compounds mycothiol Inositol Amidohydrolases N-acetyl-1-D-inosityl-2-amino-2-deoxy-alpha-D-glucopyranoside deacetylase amidase Cysteine
Topics	Amidohydrolases (chemistry, isolation & purification, metabolism) Bacterial Proteins (chemistry, isolation & purification, metabolism) Cysteine Disaccharides (biosynthesis) Glycopeptides Inositol Metalloproteins (chemistry, isolation & purification, metabolism) Mycobacterium tuberculosis (chemistry, enzymology) Pyrazoles Recombinant Proteins (chemistry, isolation & purification, metabolism) Sulfhydryl Compounds

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