Phosphoenolpyruvate carboxylase: Alteration of catalytic and regulatory properties by site-directed mutagenesis and isolation of the gene from an extreme thermophile
| Autor: | Issei Yoshioka, Kazutoyo Terada, Masato Yano, Tsutomu Nakamura, Kunimi Abe, Akio Kihara, Mamoru Takahashi, Katsura Izui |
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| Rok vydání: | 1995 |
| Předmět: |
chemistry.chemical_classification
Renewable Energy Sustainability and the Environment Allosteric regulation Mutant Mutagenesis Energy Engineering and Power Technology Biology medicine.disease_cause Pyruvate carboxylase Fuel Technology Enzyme Nuclear Energy and Engineering Biochemistry chemistry medicine Phosphoenolpyruvate carboxylase Site-directed mutagenesis Escherichia coli |
| Zdroj: | Energy Conversion and Management. 36:751-754 |
| ISSN: | 0196-8904 |
| DOI: | 10.1016/0196-8904(95)00113-r |
| Popis: | The roles of several conserved amino acid residues in phosphoenol-pyruvate carboxylase (PEPC, EC4.1.1.31) of E. coli were studied by site-directed mutagenesis. Mutant enzymes H138N (His138 replaced by Asn) and R587S lost the original catalytic activity but revealed the weak activity of HCO 3 − -dependent hydrolysis of PEP to yield pyruvate. By the use of H138N the formation of carboxyphosphate, a postulated reaction intermediate, was demonstrated for the first time. K620S and R438C were almost insensitive to an allosteric feedback inhibitor, aspartate, and the latter showed a tendency to dissociate to dimer. Furthermore, the gene for extremely thermostable PEPC was cloned and expressed in E. coli . |
| Databáze: | OpenAIRE |
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