| Autor: |
Lakshmi HP; Department of Biotechnology, Sri Venkateswara Institute of Medical Sciences, Tirupati, AP, India 517 507., Yeswanth S, Prasad UV, Vasu D, Swarupa V, Kumar PS, Narasu ML, Krishna Sarma PV |
| Jazyk: |
angličtina |
| Zdroj: |
Bioinformation [Bioinformation] 2013; Vol. 9 (4), pp. 169-73. Date of Electronic Publication: 2013 Feb 21. |
| DOI: |
10.6026/97320630009169 |
| Abstrakt: |
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 DH5α. 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±0.06mM with Hill coefficient of 1.66±0.032mM. 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. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
|
