Sodium chloride-induced modulation of the activity and thermal stability of short-chain oxidoreductase from the archaeon Thermococcus sibiricus
| Autor: | Vadim M. Gumerov, Ekaterina Yu. Bezsudnova, Vladimir Popov, T.N. Stekhanova, Andrey V. Mardanov, Serguey Y. Kleymenov, Natalya Artemova |
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| Rok vydání: | 2013 |
| Předmět: |
Circular dichroism
Protein Denaturation Molar concentration biology Sodium Temperature chemistry.chemical_element Bioengineering General Medicine Sodium Chloride biology.organism_classification Applied Microbiology and Biotechnology Biochemistry Michaelis–Menten kinetics Thermococcus Crystallography Differential scanning calorimetry chemistry Enzyme Stability Coenzyme binding Oxidoreductases Molecular Biology Biotechnology Thermostability |
| Zdroj: | Applied biochemistry and biotechnology. 171(7) |
| ISSN: | 1559-0291 |
| Popis: | Recently, we have studied properties and structural features of the thermostable halotolerant alcohol dehydrogenase from archaeon Thermococcus sibiricus (TsAdh319). In the present work, the effect of sodium chloride on activity and thermostability was explored using circular dichroism, fluorescent spectroscopy, and differential scanning calorimetry. The activity of TsAdh319 increased in the presence of NaCl and remained at the elevated level up to 4 M of NaCl. Sodium chloride at molar concentrations reduced the optimal reaction temperature, increased both Michaelis constant (K m) and k cat values for the substrates tested, decreased affinity for the coenzyme, and stoichiometry of coenzyme binding. No changes were revealed in a secondary or quaternary structure of the protein in the presence of NaCl up to 90 °C. According to differential scanning calorimetry, the irreversible unfolding started around 90 °C, the addition of NaCl decreased T m from 104.2 to 102.2 °C, and reduced ΔH from 438 to 348 kJ/mol. Kinetic studies revealed positive effect of NaCl on the TsAdh319 thermostability. The results are interpreted in regard to TsAdh319 structural data. |
| Databáze: | OpenAIRE |
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