| Summary: Ribulokinases (transferase; EC 2.7.1.16) convert L-ribuloseto L-ribulose 5 phosphateand D-ribuloseto D-ribulose 5 phosphate. Theproducts as a result of reactionscatalyzedbythisenzymeareused in pentosephosphatepathway.
Thisstudydescribesthecloning of a ribulokinase gene fromAnoxybacilluskestanbolensisAC26 Sari, isolation, andcharacterization of the gene product. A 1695 nt gene codingforribulokinasefromAnoxybacilluskestanbolensis AC26 Sarisequencedandclonedinto pET-28a(+)vector. Vectorencodingribulokinase gene wastransformedintoE.coli BL21 (DE3), expressedandpurified. Thepurifiedribulokinaseshowedoptimalactivity at 60°C in pH 9.Kmvaluefor D-ribulose was determinedtobe 0.94 mM. Effectsofmetalions (Mg2+, Ca2+, Li, K, Zn2+), β-mercaptoethanoland EDTA on theenzymeactivitywerestudied. Enzymeactivitywasincreasedby Mg2+and β-mercaptoethanol, andinhibitedbyZn2+. Inaddition, Ca2+, Li, K, and EDTA had noeffect ontheenzymeactivity. Molecularmass of A. kestanbolensis AC26 Sariribulokinasewasdeterminedto be 61.96 kDa.
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