Purification and characterization of peptide deformylase from Helicobacter pylori expressed in Escherichia coli
Main Article Content
Abstract
Helicobacter pylori is the major reason causesing gastrointestinal diseases and is characterized with the rapidly increase of multi drug resistance. In this project, we focused on studying peptide deformylase catalyzing for the crucial N-formyl cleavage from N-formyl methionyl at the beginning of new peptide chains in prokaryotes as an alternative target for screening potential anti-H. pylori compounds. The 24.5 kDa recombinant HpPDF expressed in E. coli BL21-RIL was successfully purified with a 13-fold increase in purity and 37.2% yield. HpPDF formyl removing pecific activity was determined at 145.3 U/mg with Km of 137.16 µM and kcat/Km ratio of 173.42 M-1s-1
Keywords:
Peptide deformylase, Helicobacter pylori
References
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[14] M.D. Lee, C. Antczak, Y. Li, F.M. Sirotnak, W.G. Bornmann, D.A. Scheinberg, A new human peptide deformylase inhibitable by actinonin, Biochem. Biophys. Res. Commun. 312 (2003) 309–315.
[2] Seo J.H., Woo H.O., Youn H.S. and Rhee K.H., Antibiotics resistance of Helicobacter pylori and treatment modalities in children with H. pylori infection, Korean Journal of Pediatrics, 57 2 (2014) 67.
[3] Miesel L., Greene J., Black T.A, Microbial genetics: Genetic strategies for antibacterial drug discovery. Nat Rev Genet 4 (2003) 442–456.
[4] Duckworth M.J., Okoli A.S., Mendz G.L., Novel Helicobacter pylori therapeutic targets: the unusual suspects. Expert Rev Anti Infect Ther 7 (2009) 835–867.
[5] Livingston D.M. and Leder P., Deformylation and protein synthesis, Biochemistry 8 (1969) 435.
[6] Yikun Li, Zhifeng Chen, and Weimin Gong, Enzymatic properties of a new peptide deformylase from pathogenic bacterium Leptospira interrogans, Biochemical and Biophysical Research Communications 295 (2002) 884–889.
[7] Kumar A., Nguyen K.T., Srivathsan S., Ornstein B., Turley S., Hirsh I., Pei D.,
and Hol W.G.J, Crystals of Peptide Deformylase from Plasmodium falciparum Reveal Critical Characteristics of the Active Site for Drug Design. Structure, 10 (2002) 357–367.
[8] Clements J.M., Beckett R.P., Brown A., Catlin G., Lobell M., Palan S., Thomas W., Whittaker M., Wood S., Salama S., Baker P.J, Rodgers H.F., Barynin V., Rice D.W., and Hunter M.G., Antibiotic Activity and Characterization of BB-3497, a Novel Peptide Deformylase inhibitor antimicrobial agents and chemotherapy 45 2 (2002) 563–570.
[9] Đỗ Thị Thanh Trung, Lê Hồng Điệp, Phạm Bảo Yên, Nhân dòng và bước đầu biểu hiện peptide deformylase của chủng vi khuẩn Helicobacter pylori phân lập ở Việt Nam. Tạp chí Khoa học ĐHQGHN: Khoa học Tự nhiên và Công nghệ, 31 4S (2015) 453-460.
[10] Lazennec C. and Meinnel T., Formate Dehydrogenase-Coupled Spectrophotometric Assay of Peptide Deformylase. Anal Biochem 244 (1997) 180–182.
[11] Meinnel T. and Blanquet S., Enzymatic Properties of Escherichia coli Peptide Deformylase. J Bacteriol 177 7 (1995) 1883–1887.
[12] Rajagopalan P.T., Datta A. and Pei D., Purification, characterization, and inhibition of peptide deformylase from Escherichia coli, Biochemistry, 36 (1997) 13910.
[13] Han C., Wang Q., Dong L., Sun H., Peng S., Chen J., Yang Y., Yue J., Shen X. and Jiang H., Molecular cloning and characterization of a new peptide deformylase from human pathogenic bacterium Helicobacter pylori, Biochemical and Biophysical Research Communications, 319 (2004) 1292.
[14] M.D. Lee, C. Antczak, Y. Li, F.M. Sirotnak, W.G. Bornmann, D.A. Scheinberg, A new human peptide deformylase inhibitable by actinonin, Biochem. Biophys. Res. Commun. 312 (2003) 309–315.