Thermostable farnesyl diphosphate synthase of Bacillus stearothermophilus: crystallization and site-directed mutagenesis
| Autor: | Tanetoshi Koyama, Ayumi Takeshita, Kazuhiro Saito, Kyozo Ogura, Tokuzo Nishino, Masami Osabe, Shusei Obata |
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| Rok vydání: | 1994 |
| Předmět: |
Hot Temperature
Molecular Sequence Data Restriction Mapping Mutant Arginine medicine.disease_cause General Biochemistry Genetics and Molecular Biology Geobacillus stearothermophilus Serine Farnesyl diphosphate synthase Valine Enzyme Stability medicine Humans Amino Acid Sequence Cloning Molecular Site-directed mutagenesis Escherichia coli Conserved Sequence Base Sequence Sequence Homology Amino Acid ATP synthase biology Chemistry Fungi Dimethylallyltranstransferase Recombinant Proteins Kinetics Oligodeoxyribonucleotides Biochemistry Mutagenesis Site-Directed biology.protein Thermodynamics Electrophoresis Polyacrylamide Gel Crystallization Plasmids Cysteine |
| Zdroj: | Acta Biochimica Polonica. 41:281-292 |
| ISSN: | 1734-154X 0001-527X |
| DOI: | 10.18388/abp.1994_4717 |
| Popis: | The gene for thermostable farnesyl diphosphate synthase from Bacillus stearothermophilus was cloned, sequenced, and overexpressed in Escherichia coli. The synthase was purified to homogeneity and crystallized. The enzyme carried only two cysteine residues in contrast to its counterparts from other sources, which have four to six cysteine residues. Either or both of the cysteine residues can be replaced with serine without causing a loss of the catalytic activity. The conserved arginine residue that occupies the third position from the C-terminus was also replaced with valine without significant loss of activity, but the valine mutant showed a weakened affinity for isopentenyl diphosphate. |
| Databáze: | OpenAIRE |
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