%0 Journal Article
%T Cloning, expression and characterization of glucokinase gene involved in the glucose-6-phosphate formation in Staphylococcus aureus
%A Hanumanthu Prasanna Lakshmi
%A Sthanikam Yeswanth
%A Uppu Venkateswara Prasad
%A Dudipeta Vasu
%J Bioinformation
%D 2013
%I 
%X Glucose-6-phosphate (G-6-P) formation in Staphylococcus aureus is catalysed by glucokinase (glkA) gene under high glucose concentration leading to upregulation of various pathogenic factors; therefore the present study is aimed in the cloning and characterization of glk A gene from S. aureus ATCC12600. The glk A gene was cloned in the Sma I site of pQE 30, sequenced (Accession number: JN645812) and expressed in E. coli DH5a. The recombinant glk A expressed from the resultant glk A 1 clone was purified using nickel metal chelate chromatography, the pure enzyme gave single band in SDS-PAGE with molecular weight of 33kDa. The rglk A showed very high affinity to glucose Km 5.1¡À0.06mM with Hill coefficient of 1.66¡À0.032mM. Analysis of glucokinase sequence of S. aureus showed presence of typical ATP binding site and ROK motif CNCGRSGCIE. Sequentially and phylogenetically S. aureus glk A exhibited low identity with other bacterial glk A and 21% homology with human glucokinase (GCK). Functionally, S. aureus glk A showed higher rate of G-6-P formation compared to human GCK which may have profound role in the pathogenesis.
%K glk A
%K pQE 30
%K ROK
%K Km
%U http://bioinformation.net/009/97320630009169.htm