Expression, Purification, Characteristics and Homology Modeling of the HMGS from Streptococcus pneumoniae
| Autor: | Qingye Zhang, Chen Li, Jian Wan, Rui Han, Gu-Zhen Cui, Ya-Li Ben, Jie Zhang, Deli Liu |
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| Rok vydání: | 2009 |
| Předmět: |
Hydroxymethylglutaryl-CoA Synthase
Models Molecular Protein Conformation Health Toxicology and Mutagenesis Coenzyme A Molecular Sequence Data medicine.disease_cause Enterococcus faecalis law.invention chemistry.chemical_compound Affinity chromatography law medicine Amino Acid Sequence Homology modeling Cloning Molecular Escherichia coli chemistry.chemical_classification Base Sequence biology Public Health Environmental and Occupational Health Gene Expression Regulation Bacterial biology.organism_classification Streptococcus pneumoniae Enzyme chemistry Biochemistry Mutation Recombinant DNA Mevalonate pathway |
| Zdroj: | Biomedical and Environmental Sciences. 22:229-236 |
| ISSN: | 0895-3988 |
| DOI: | 10.1016/s0895-3988(09)60050-7 |
| Popis: | 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 μmol/min/mg. Optimal conditions were pH 9.75 and 10 mmol/L MgCl2 at 37 ℃. The Vmax and Km were 4.69 μmol/min/mg and 213 μmol/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. |
| Databáze: | OpenAIRE |
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