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Biomed Environ Sci ; 22(3): 229-36, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19725466

ABSTRACT

OBJECTIVE: 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). METHODS: 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. RESULTS: 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. CONCLUSION: 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.


Subject(s)
Gene Expression Regulation, Bacterial/physiology , Hydroxymethylglutaryl-CoA Synthase/chemistry , Hydroxymethylglutaryl-CoA Synthase/metabolism , Streptococcus pneumoniae/enzymology , Amino Acid Sequence , Base Sequence , Cloning, Molecular , Hydroxymethylglutaryl-CoA Synthase/genetics , Models, Molecular , Molecular Sequence Data , Mutation , Protein Conformation , Streptococcus pneumoniae/genetics
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