Expression, purification, characteristics and homology modeling of the HMGS from Streptococcus pneumoniae / 生物医学与环境科学(英文)
Biomedical and Environmental Sciences
;
(12): 229-236, 2009.
Article
in English
| WPRIM
| ID: wpr-360671
ABSTRACT
<p><b>OBJECTIVE</b>To understand the molecular basis for a potential reaction mechanism and develop novel antibiotics with homology modeling for 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) synthase (HMGS).</p><p><b>METHODS</b>The genetic engineering technology and the composer module of SYBYL7.0 program were used, while the HMGS three-dimensional structure was analyzed by homology modeling.</p><p><b>RESULTS</b>The mvaS gene was cloned from Streptococcus pneumoniae and overexpressed in Escherichia coli from a pET28 vector. The expressed enzyme (about 46 kDa) was purified by affinity chromatography with a specific activity of 3.24 micromol/min/mg. Optimal conditions were pH 9.75 and 10 mmol/L MgCl2 at 37 degrees C. The V(max) and K(m) were 4.69 micromol/min/mg and 213 micromol/L respectively. The 3D model of S. pneumoniae HMGS was established based on structure template of HMGS of Enterococcus faecalis.</p><p><b>CONCLUSION</b>The structure of HMGS will facilitate the structure-based design of alternative drugs to cholesterol-lowering therapies or to novel antibiotics to the Gram-positive cocci, whereas the recombinant HMGS will prove useful for drug development against a different enzyme in the mevalonate pathway.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Physiology
/
Protein Conformation
/
Streptococcus pneumoniae
/
Molecular Sequence Data
/
Base Sequence
/
Models, Molecular
/
Gene Expression Regulation, Bacterial
/
Chemistry
/
Amino Acid Sequence
/
Cloning, Molecular
Type of study:
Prognostic study
Language:
English
Journal:
Biomedical and Environmental Sciences
Year:
2009
Type:
Article
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