Characterization of a novel esterase Rv0045c from Mycobacterium tuberculosis
| Autor: | Xiangdong Zheng, Hai Pang, Jiubiao Guo, Tingyi Wen, Lipeng Xu, Kehui Xu, Siguo Liu, Shentao Li, Zhongyuan Liu |
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| Rok vydání: | 2010 |
| Předmět: |
Circular dichroism
Science Biochemistry/Biocatalysis Esterase Biochemistry Protein Structure Secondary Protein structure Biochemistry/Protein Chemistry Hydrolase Escherichia coli Biochemistry/Macromolecular Chemistry Peptide sequence Protein secondary structure chemistry.chemical_classification Multidisciplinary Chemistry Esterases Mycobacterium tuberculosis Chromatography Ion Exchange Molecular biology Enzyme structure Recombinant Proteins Enzyme Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Chromatography Gel Medicine Electrophoresis Polyacrylamide Gel Research Article |
| Zdroj: | PLoS ONE PLoS ONE, Vol 5, Iss 10 (2010) |
| ISSN: | 1932-6203 |
| Popis: | BackgroundIt was proposed that there are at least 250 enzymes in M. tuberculosis involved in lipid metabolism. Rv0045c was predicted to be a hydrolase by amino acid sequence similarity, although its precise biochemical characterization and function remained to be defined.Methodology/principal findingsWe expressed the Rv0045c protein to high levels in E. coli and purified the protein to high purity. We confirmed that the prepared protein was the Rv0045c protein by mass spectrometry analysis. Circular dichroism spectroscopy analysis showed that the protein possessed abundant β-sheet secondary structure, and confirmed that its conformation was stable in the range pH 6.0-10.0 and at temperatures ≤ 40 °C. Enzyme activity analysis indicated that the Rv0045c protein could efficiently hydrolyze short chain p-nitrophenyl esters (C₂-C₈), and its suitable substrate was p-nitrophenyl caproate (C₆) with optimal catalytic conditions of 39 °C and pH 8.0.Conclusions/significanceOur results demonstrated that the Rv0045c protein is a novel esterase. These experiments will be helpful in understanding ester/lipid metabolism related to M. tuberculosis. |
| Databáze: | OpenAIRE |
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