Ribokinase from E. coli: Expression, purification, and substrate specificity.
 |
Related Articles |
Ribokinase from E. coli: Expression, purification, and substrate specificity.
Bioorg Med Chem. 2006 Jun 17;
Authors: Chuvikovsky DV, Esipov RS, Skoblov YS, Chupova LA, Muravyova TI, Miroshnikov AI, Lapinjoki S, Mikhailopulo IA
Ribokinase (RK) was expressed in the Escherichia coli ER2566 cells harboring the constructed expression plasmid encompassing the rbsK gene, encoding ribokinase. The recombinant enzyme was purified from sonicated cells by double chromatography to afford a preparation that was ca. 90% pure and had specific activity of 75mumol/minmg protein. Catalytic activity of RK: (i) is strongly dependent on the presence of monovalent cations (potassium>>>ammonium>cesium), and (ii) is cooperatively enhanced by divalent magnesium and manganese ions. Besides d-ribose and 2-deoxy-d-ribose, RK was found to catalyze the 5-O-phosphorylation of d-arabinose, d-xylose, and d-fructose in the presence of ATP, and potassium and magnesium ions; l-ribose and l-arabinose are not substrates for the recombinant enzyme. A new radiochemical method for monitoring the formation of d-pentofuranose-5-[(32)P]phosphates in the presence of [gamma-(32)P]ATP and RK is reported.
PMID: 16784868 [PubMed - as supplied by publisher]
Related posts:
- Cloning, expression, purification and characterization of recombinant (+)-germacrene D synthase from Zingiber officinale.
- In vitro refolded napin-like protein of Momordica charantia expressed in Escherichia coli displays properties of native napin.
- Expression, purification, crystallization and preliminary X-ray analysis of strictosidine glucosidase, an enzyme initiating biosynthetic pathways to a unique diversity of indole alkaloid skeletons.
- Purification, molecular cloning, and cell-specific gene expression of the alkaloid-accumulation associated protein CrPS in Catharanthus roseus.
- The P4 metal binding site in RNase P RNA affects active site metal affinity through substrate positioning.