Cloning, purification and characterization of geobacillus stearothermophilus v uroporphyrinogen-iii c-methyltransferase: evaluation of its role in resistance to potassium tellurite in escherichia coli
| Autor: | Claudia P. Saavedra, Radhika Burra, Manuel A. Araya, Iván L. Calderón, Derie E. Fuentes, Claudio C. Vásquez, Thomas G. Chasteen, José M. Pérez, Juan C. Tantaleán |
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| Jazyk: | angličtina |
| Rok vydání: | 2009 |
| Předmět: |
S-Adenosylmethionine
Molecular Sequence Data Uroporphyrinogens Biology Molecular cloning medicine.disease_cause Microbiology Geobacillus stearothermophilus chemistry.chemical_compound Uroporphyrinogen medicine Escherichia coli Amino Acid Sequence Cloning Molecular Molecular Biology chemistry.chemical_classification General Medicine Methyltransferases biology.organism_classification Enterobacteriaceae Tellurate Recombinant Proteins Enzyme chemistry Biochemistry Uroporphyrinogen III Mutagenesis Site-Directed Tellurium |
| Zdroj: | RESEARCH IN MICROBIOLOGY Artículos CONICYT CONICYT Chile instacron:CONICYT |
| Popis: | The Geobacillus stearothermophilus V cobA gene encoding uroporphyrinogen-III C-methyltransferase (also referred to as SUMT) was cloned into Escherichia coli and the recombinant enzyme was overexpressed and purified to homogeneity. The enzyme binds S-adenosyl-L-methionine and catalyzes the production of III methyl uroporphyrinogen in vitro. E. coli cells expressing the G. stearothermophilus V cobA gene exhibited increased resistance to potassium tellurite and potassium tellurate. Site-directed mutagenesis of cobA abolished tellurite resistance of the mesophilic, heterologous host and SUMT activity in vitro. No methylated, volatile derivatives of tellurium were found in the headspace of tellurite-exposed cobA-expressing E. coli, suggesting that the role of SUMT methyltransferase in tellurite(ate) detoxification is not related to tellurium volatilization. |
| Databáze: | OpenAIRE |
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