Pakistan Journal of Biological Sciences1028-88801812-5735Asian Network for Scientific Information10.3923/pjbs.2020.561.566Escherichia coli BL21 (DE3) pET21b dpe]]>EdyLianiDeby YurnalizaYurnaliza SaksonoBudi 42020234Background and Objective: The DPEase enzyme from Agrobacterium tumefaciens is more efficient and has a high activity in D-fructose. The dpe gene has been successfully cloned to Escherichia coli BL21 (DE3) pET-21b dpe but the enzyme has not been purified and its character is unknown. The intent of this study was to purify and assign of DPEase enzyme by recombinant E. coli. Materials and Methods: The enzyme was clarified by affinity chromatography and then characterized by following pH, temperature, co-factor parameters. Analysis of molecular weight proteins was done by SDS-PAGE. Results: Through purification, the purified DPEase activity was increased 1,01 times than crude and with 84.2% of yield. The DPEase had an the maximum temperature is 40°C and pH was 8.5. The presence of Mg2+, Mo2+, Cu2+, Ca2+ and Zn2+ inhibited the activity of the enzyme while of Co2+, Mn2+, Fe2+, Ni2+ enhanced the activity. Estimation of molecular weight through SDS-PAGE revealed that weight of DPEase was 32 kDa. Conclusion: Purified DPease enzymes shows clear bands that demonstrate successful purification using affinity chromatography. It is expected that after pure enzymes are obtained the character of the enzymes working will be maximized.]]>Miller, B.S. and T. Swain,196011344348Hossain, M.A., S. Kitagaki, D. Nakano, A. Nishiyama and Y. Funamoto et al.,2011405712Zunino, S.J.,20091391794S1800SHossain, A., F. Yamaguchi, T. Matsunaga, Y. Hirata and K. Kamitori et al.,2012425717723Matsuo, T., H. Suzuki, M. Hashiguchi and K. Izumori,2002487780Hayashi, N., T. Iida, T. Yamada, K. Okuma and I. Takehara et al.,201074510519FDA.,20112011Mu, W., W. Zhang, Y. Feng, B. Jiang and L. Zhou,20129414611467Jia, M., W. Mu, F. Chu, X. Zhang, B. Jiang, L.L. Zhou and T. Zhang,2014Clostridium bolteae for D-psicose production: Cloning, expression, purification and characterization.]]>98717725Chen, X., W. Wang, J. Xu, Z. Yuan and T. Yuan et al.,20171051823Kim, K., H.J. Kim, D.K. Oh, S.S. Cha and S. Rhee,2006Agrobacterium tumefaciens and its complex with true substrate D-fructose: A pivotal role of metal in catalysis, an active site for the non-phosphorylated substrate and its conformational changes.]]>361920931Zhang, W., D. Fang, Q. Xing, L. Zhou, B. Jiang and W. Mu,2013Clostridium scindens 35704.]]>2013Zhu, Y., Y. Men, W. Bai, X. Li, L. Zhang, Y. Sun and Y. Ma,2012Ruminococcus sp. in Escherichia coli and its potential application in d-psicose production.]]>3419011906Choi, J.G., Y.H. Ju, S.J. Yeom and D.K. Oh,2011Agrobacterium tumefaciens by random and site-directed mutagenesis.]]>7773167320Indradewi, R.,2017Agrbacterium tumafaciens sebagai gen penyandi enzim D-psicose 3-Epimerase (DPEase).]]>2017Kierstan, M.P.J.,197820447450Lowry, O.H., N.J. Rosebrough, A.L. Farr and R.J. Randall,1951193265275Gräslund, S., P. Nordlund, J. Weigelt, B.M. Hallberg and J. Bray et al.,20085135146Oh, D.K., N.H. Kim, H.J. Kim, C.S. Park and S.W. Kim et al.,2007Sinorhizobium sp.]]>2007Robinson, P.K.,201559141Lim, B.C., H.J. Kim and D.K. Oh,200944822828Park, C.S., C.S. Park, K.C. Shin and D.K. Oh,2016Agrobacterium tumefaciens.]]>121186190Tseng, W.C., C.N. Chen, C.T. Hsu, H.C. Lee and H.Y. Fang et al.,2018Agrobacterium sp. ATCC 31749 and identification of an important interfacial residue.]]>112767774