التفاصيل البيبلوغرافية
| العنوان: |
NAD+-Dependent Formate Dehydrogenase from Themotolerant Yeast Ogataea parapolymorpha: Properties and Protein Engineering of the N-Terminal Sequence. |
| المؤلفون: |
Pometun, Anastasia A., Shaposhnikov, Leonid A., Zubanova, Sofiya A., Kovalevskii, Rostislav P., Atroshenko, Denis L., Pometun, Evgenii V., Savin, Svyatoslav S., Tishkov, Vladimir I. |
| المصدر: |
Biochemistry (00062979); Sep2023, Vol. 88 Issue 9, p1378-1389, 12p |
| مصطلحات موضوعية: |
PROTEIN engineering, AMINO acid residues, NAD (Coenzyme), N-terminal residues, YEAST, THERMAL stability, DEHYDROGENASES, SIRTUINS |
| مستخلص: |
Previously, the gene of formate dehydrogenase (FDH, EC 1.2.1.2) from the thermotolerant methylotrophic yeast Ogataea parapolymorpha DL 1 (OpaFDH) was cloned in our laboratory. Recombinant enzyme with additional glycine amino acid residue (OpaFDH_GK) was obtained in Escherichia coli cells in active and soluble form with a yield of more than 1 g per liter of the medium. In the present work, a detailed comparison of this enzyme with FDHs from other sources was carried out. Among eukaryotic formate dehydrogenases, OpaFDH has the highest thermal stability. To elucidate effect of N-terminal residue on the properties of the enzyme, OpaFDH_K (identical to natural) and OpaFDH_AK variants containing an additional Ala residue at the N-terminus were also obtained. It was shown that addition of an Ala residue to the N-terminus reduces four-fold the rate constant of thermal inactivation compared with the addition of a Gly residue. Addition of six more histidine residues to the N-terminus of OpaFDH_AK leads to acceleration of purification, practically does not affect kinetic parameters, but somewhat reduces thermal stability, which, however, can be restored to the level of OpaFDH_AK stability by adding 0.5 M NaCl. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |