Pakistan Journal of Biological Sciences1028-88801812-5735Asian Network for Scientific Information10.3923/pjbs.2016.106.114in vitro Analysis of Its Anticancer Efficacy on Different Human Cancer Cell Lines]]>MuharramM.M. 32016193Recombinant methioninase (rMETase) is an enzyme that has antitumor activity. In this work, METase gene from Pseudomonas putida ATTCC 8209 was cloned to pT7-7 plasmid (yielded, PT7-METase-R7 clone) and expressed in E. coli strain BL21 (DE3). A protein band with a molecular massof 42 kDa was visualized by SDS-PAGE. The applied protocol yielded a total protein of 3.13 g with a recovery of 66.89% and a specific activity of 18.59 U mg1 which considered as a low yield. However, when the METase gene was cloned to the vector (pTrc99A, clone: pTrc99A-MET-3) cells of E. coli JM109 yielded a total protein of 32.63 g with a recovery of 41.62% and a specific activity of 54.86 U mg1 which revealed that the enhancement of METase gene expression by trc promoter was more than the T7 RNA polymerase promoter. The t1/2 of the rMETase was 2 h asanalyzed in mice by IV injection. Antitumor efficacy of rMETase was studied in five human cancer cell lines. At 1 U mL1 the growth rate of treated colon cancer cell lines, Colo205 and SW620, with rMETase was 46 and 32% relative to control, respectively. With the ovarian cancer cell line (A2780) rMETase produced an inhibition effect of 54% at 1.5 U mL1. In addition, the growth rate was reduced to 45 and 53% with the skin cancer cell line (A375) and the breast cancer cell line (MCF-7), respectively. These results indicate the feasibility of rMETase for use as a potent antitumor agent.]]>Anderson, M.E.,1998111-112114Bradford, M.M.,197672248254Cooper, A.J.L.,198352187222El-Sayed, A.S.A.,2009Aspergillus flavipes under solid-state fermentation.]]>49331341El-Sayed, A.S.A.,201086445467Esaki, N. and K. Soda,1987Pseudomonas putida and Aeromonas.]]>143459465Ghoshal, A.K., D.S. Sarma and E. Farber,19861986pp: 283-292pp: 283-292Hoffman, R.M.,19847384987Hoffman, R.M.,2015152131Hori, H., K. Takabayashi, L. Orvis, D.A. Carson and T. Nobori,1996Pseudomonas putida L-methionine-α-deamino-γ-mercaptomethane-lyase.]]>5621162122Howarth, G.S. and H. Wang,201355881Hu, J. and N.K.V. Cheung,2009In vitro and in vivo efficacy against neuroblastoma and its synergism with chemotherapeutic drugs.]]>12417001706Huang, K.Y., H.Y. Hu, Y.L. Tang, F.G. Xia, X.Q. Luo and J.Z. Liu,20151354925507Inoue, H., K. Inagaki, M. Sugimoto, N. Esaki, K. Soda and H. Tanaka,1995Pseudomonas putida.]]>11711201125Inoue, H., K. Inagaki, N. Adachi, T. Tamura, N. Esaki, K. Soda and H. Tanaka,2000Pseudomonas putida.]]>6423362343Kokkinakis, D.M., S.C. Schold Jr., H. Hori and T. Nobori,199729195204Kreis, W. and M. Goodenow,19783822592262Laemmli, U.K.,1970227680685Lishko, V.K., O.V. Lishko and R.M. Hoffman,19931314651468Lu, S. and D.E. Epner,200038123130Megosh, L., S.K. Gilmour, D. Rosson, A.P. Soler, M. Blessing, J.A. Sawicki and T.G. O'Brien,19955542054209Motoshima, H., K. Inagaki, T. Kumasaka, M. Furuichi and H. Inoue et al.,2000Pseudomonas putida.]]>128349354Palwai, N.R., X.P. Zang, R.G. Harrison, D. Benbrook and J.T. Pento,2009L-methioninase-containing fusion protein targeted to the urokinase receptor.]]>84271275Sambrook, J., E.F. Fritsch and T.A. Maniatis,19892nd Edn.,Pages: 397Pages: 397Sinha, R., T.K. Cooper, C.J. Rogers, I. Sinha and W.J. Turbitt et al.,20147416631673Stabler, S.P., P.D. Marcell, E.R. Podell and R.H. Allen,1987162185196Stone, E., O. Paley, J. Hu, B. Ekerdt, N.K. Cheung and G. Georgiou,2012718221829Strekalova, E., D. Malin, D.M. Good and V.L. Cryns,20152127802791Takakura, T., K. Mitsushima, S. Yagi, K. Inagaki and H. Tanaka et al.,2004327233240Takakura, T., T. Ito, S. Yagi, Y. Notsu and T. Itakura et al.,200670183192Tan, Y., X. Sun, M. Xu, X. Tan and A. Sasson et al.,1999521572163Tan, Y., M. Xu, X. Tan, X. Tan and X. Wang et al.,19979233245Tan, Y., M. Xu, H. Guo, X. Sun, T. Kubota and R.M. Hoffman,1996in vivo.]]>1639313936Tanaka, H., N. Esaki and K. Soda,1977Pseudomonas ovalis.]]>16100106Tanaka, H., N. Esaki, T. Yamamoto and K. Soda,1976Pseudomonas ovalis.]]>66307311Thomas, T. and T.J. Thomas,20037113126Towbin, H., T. Staehelin and J. Gordon,19797643504354Van Rite, B.D., Y.A. Lazrak, M.L. Pagnon, N.R. Palwai, L.F. Neves, P.S. McFetridge and R.G. Harrison,2011301177184Yang, Z., J. Wang, Q. Lu, J. Xu and Y. Kobayashi et al.,20046466736678Yano, S., S. Li, Q. Han, Y. Tan, M. Bouvet, T. Fujiwara and R.M. Hoffman,2014587298736Yoshioka, T., T. Wada, N. Uchida, H. Maki and H. Yoshida et al.,1998in vivo and in vitro, synergy with 5-fluorouracil and safety of recombinant methioninase.]]>5825832587Zhao, R., F.E. Domann and W. Zhong,2006532753284