Characterization of a novel exported esterase Rv3036c from Mycobacterium tuberculosis

• Published in: Protein expression and purification 2014-12, Vol.104, p.50-56
• Main Authors: Chen, Liping, Dang, Guanghui, Deng, Xiaoxia, Cao, Jun, Yu, Shenye, Wu, Defeng, Pang, Hai, Liu, Siguo
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Enzyme Activation; Escherichia coli; Esterases - biosynthesis; Esterases - isolation & purification; Exported esterase; Mycobacterium tuberculosis; Mycobacterium tuberculosis - enzymology; Nitrophenols - metabolism; Protein Sorting Signals; rv3036c
• ISSN: 1046-5928; 1096-0279
• DOI: 10.1016/j.pep.2014.09.003

•The esterase motif SXXK was identified in Rv3036c from M. tuberculosis.•A highly pure soluble recombinant Rv3036c was obtained.•Rv3036c showed hydrolase activity for water-soluble esters.•Rv3036c was present at the surface of mycobacteria. Mycobacterium tuberculosis possesses an unusually high number of genes involved in the metabolism of lipids. Driven by a newly described esterase motif SXXK in the amino acid sequence and a predicted signal peptide, the gene rv3036c from M. tuberculosis was cloned and characterized biochemically. Rv3036c efficiently hydrolyzes soluble p-nitrophenyl esters but not emulsified lipid. The highest activity of this enzyme was observed when p-nitrophenyl acetate (C2) was used as the substrate. Based on the activities, Rv3036c was classified as a nonlipolytic hydrolase. The results of immunoreactivity studies on the subcellular mycobacterial fractions suggested that the enzyme was present in the cell wall and cell membrane in mycobacteria. In summary, Rv3036c was characterized as a novel cell wall-anchored esterase from M. tuberculosis.