title: Characterization of a Mn-dependent fructose-1,6-bisphosphate aldolase in Deinococcus radiodurans.
abstract: The key enzyme of the glycolytic pathway of Deinococcus radiodurans, fructose-1,6-bisphosphate aldolase, could be induced independently by glucose and Mn. The enzyme exhibited the characteristics of the metal-dependent Class II aldolases. Unlike most Class II aldolases, the deinococcal aldolase preferred Mn, not Zn, as a cofactor. The fbaA gene encoding the deinococcal aldolase was cloned and the protein overproduced in various Escherichia coli expression hosts. However, the overexpressed deinococcal enzyme aggregated and formed inclusion bodies. Dissolving these inclusion bodies by urea and subsequent purification by nickel affinity chromatography, resulted in a protein fraction that exhibited aldolase activity only in the presence of Mn. This active aldolase fraction exhibited masses of about 70 kDa and 35 kDa by gel filtration and by SDS gel electrophoresis, respectively, suggesting that the active aldolase was a dimer.
authors.affiliation: Department of Microbiology and Molecular Cell Sciences, University of Memphis, Memphis, TN 38152, USA.
authors.forename: Yong-Mei
authors.initials: YM
authors.lastname: Zhang
authors.forename: Jong-Kang
authors.initials: JK
authors.lastname: Liu
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Shouri
authors.forename: Tit-Yee
authors.initials: TY
authors.lastname: Wong
mesh_terms: D002850:Chromatography, Gel / Q000379:methods; D034301:Deinococcus / Q000201:enzymology* / Q000254:growth & development; D019281:Dimerization; D004789:Enzyme Activation / Q000187:drug effects; D005634:Fructose-Bisphosphate Aldolase / Q000096:biosynthesis* / Q000737:chemistry* / Q000187:drug effects; D008345:Manganese / Q000737:chemistry* / Q000378:metabolism; D008970:Molecular Weight; D012680:Sensitivity and Specificity; D013329:Structure-Activity Relationship; D014508:Urea / Q000494:pharmacology
chemical_list: D008345:Manganese; D014508:Urea; D005634:Fructose-Bisphosphate Aldolase