Identification and characterization of serine acetyltransferase encoded by the Mycobacterium tuberculosis Rv2335 gene

Qiu,Juanjuan; Wang,Daiqing; Ma,Yufang; Jiang,Tao; Xin,Yi

doi:10.3892/ijmm.2013.1298

May 2013 Volume 31 Issue 5

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May 2013 Volume 31 Issue 5

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Article

Identification and characterization of serine acetyltransferase encoded by the Mycobacterium tuberculosis Rv2335 gene

Authors:
- Juanjuan Qiu
- Daiqing Wang
- Yufang Ma
- Tao Jiang
- Yi Xin
View Affiliations / Copyright

Affiliations: Department of Biotechnology, Dalian Medical University, Dalian 116044, P.R. China, Department of Biochemistry and Molecular Biology, Dalian Medical University, Dalian 116044, P.R. China
Pages: 1229-1233
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Published online on: March 12, 2013

https://doi.org/10.3892/ijmm.2013.1298
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Abstract

Serine acetyltransferase (CysE) is the first enzyme involved in the two-step enzymatic pathway of L-cysteine biosynthesis in bacteria and plants, but not in humans. CysE catalyzes the biosynthesis of O-acetyl-L-serine and CoA from L-serine (L-Ser) and acetyl-CoA (AcCoA). Mycobacterium tuberculosis (M. tuberculosis) Rv2335 was predicted as the cysE gene encoding serine acetyltransferase. In this study, the M. tuberculosis Rv2335 gene was cloned and the CysE protein was expressed in E. coli BL21 (DE3). The M. tuberculosis CysE protein was purified by Ni2+ affinity chromatography and confirmed by SDS-PAGE, western blotting and mass spectrometry. The serine acetyltransferase activity of the M. tuberculosis CysE protein was detected using Ellman's reagent. M. tuberculosis CysE displayed optimal activity at pH 7.5 and 37˚C. The Michaelis constant for AcCoA and L-Ser was 0.0513±0.0050 and 0.0264±0.0006 mM, respectively. The maximum velocity (Vmax) for CysE was 0.0073±0.0005 mM/min. The CysE assay and the determination of the kinetic parameters of M. tuberculosis CysE may be helpful for screening its inhibitors in anti-tuberculosis drug discovery.

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Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Identification and characterization of serine acetyltransferase encoded by the Mycobacterium tuberculosis Rv2335 gene

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