Asian Journal of Biotechnology1996-0700xxxx-xxxxKnowledgia Review10.3923/ajbkr.2011.478.485Pseudomonas aeruginosa]]>VijayaraghavanPonnuswamy BrightSurgen A. Nishanth LiptonAnuj VincentS.G. Prakash 5201135The aim of this study was to purify and characterize citric acid cycle enzyme malate dehydrogenase (MDH; EC 1. 1. 1. 37) from Pseudomonas aeruginosa. The purification steps consisted of ammonium sulphate precipitation, ion-exchange chromatography and gel filtration. A typical procedure provided 638-fold purification with 23.0% yield. Single band was observed in both native gradient-and SDS-PAGE. The molecular weight estimated for the native enzyme was 70.5 kDa whereas subunit values of 36 kDa were determined. Hence, MDH is a dimer of identical subunits. The enzyme was highly active at pH 8.0 when NADH was used as the cofactor and was highly stable at pH 7.0. The optimum temperature for the enzyme activity was recorded to be 40°C. Oxaloacetate was determined as the specific substrate with an apparent km of 10 μM. The characteristics of thermo-stability and its high activity at alkaline pH suggest its potential diagnostic, therapeutic and beverage related applications. This MDH may be of value in developing a serological test for pneumonia which is caused by P. aeruginosa.]]>Blum, H., H. Beier and H.J. Gross,198789399Chaffai, R. and E.El-Ferjani,2005Zea mays L.).]]>16473Charnock, C., U.H. Refseth and R. Sirevag,1992Chlorobium vibrioforme, Chlorobium tepidum and Heliobacterium gestii: Purification, characterization and investigation of dinucleotide binding by dehydrogenases by use of empirical methods of protein sequence analysis.]]>17413071313Eprintsev, A.T., M.I. Falaleeva, I.Y. Stepanova and N.V. Parfenova,2003Beggiatoa.]]>68172176Eprintsev, A.T., M.I. Falaleeva, I.Y. Stepanova, N.V. Parfenova and M. Zuzu,2003Beggiatoa leptomitiformis.]]>30243247Eprintsev, A.T., M.I Falaleeva and N.V. Parfenova,2005Vulcanithermus medioatlanticus.]]>7010271030Farfan, M., D. Minana-Galbis, A. Garreta, J.G. Loren and M.C. Fuste,2010Aeromonas.]]>33427435Ge, Y.D., Z.Y. Cao, Z.D. Wang, L.L. Chen, Y.M. Zhu and G.P. Zhu,2010Streptomyces coelicolor.]]>7421942201Genda, T., T. Nakamatsu and H. Ozak,2003Corynebacterium glutamicum.]]>95562566Goward, C.R. and D.J. Nicholls,1994318831888Labrou, N.E. and Y.D. Clonis,1997Pseudomonas stutzeri: Purification and characterization.]]>337103114Lowry, O.H., N.J. Rosebrough, A.L. Farr and R.J. Randall,1951193265275Masuko, T., R. Takahashi and T. Tokuyama,1997505159Minarik, P., N. Tomaskova, M. Kollarova and M. Antalik,200221257265Mori, H., A. Yamashila and S. Maitani,200753128131Nishizawa, H., N. Kita, S. Okimura, E. Takao and Y. Abe,19889803806Oikawa, T., N. Yamamoto, K. Shimoke, S. Uesato, T. Ikeuchi and T. Fujioka,2005Flavobacterium frigidimaris KUC-1.]]>6921462154Roderick, S.L. and L.J. Banaszak,198626194619464Shinagawa, E., T. Fujishima, D. Moonmangmee, G. Theeragool, H. Toyama, K. Matsushita and O. Adachi,2002Acetobacter sp. SKU 14.]]>66298306Van-Kuijk, B.L.M. and A.J.M. Stams,1996144141144Walk, R.A. and B. Hock,1977Citrullus vulgaris) cotyledons. II Kinetic properties of the purified isoenzymes.]]>136221228Yennaco, L.J., Y. Hu and J.F. Holden,2007Pyrobaculum islandicum.]]>11741746You, K.S. and N.O. Kaplan,1975Pseudomonas testosteroni.]]>123704716Yueh, A.Y., C.S. Chung and Y.K. Lai,1989Nitzschia alba.]]>258221228Laemmli, U.K.,1970227680685