Details

Autor(en) / Beteiligte

• Cheng, Zhongyi
• Lan, Yao
• Guo, Junling
• Ma, Dong
• Jiang, Shijin
• Lai, Qianpeng
• Zhou, Zhemin
• Peplowski, Lukasz

Titel

Computational Design of Nitrile Hydratase from Pseudonocardia thermophila JCM3095 for Improved Thermostability

Ist Teil von

• Molecules (Basel, Switzerland), 2020-10, Vol.25 (20), p.4806

Ort / Verlag

Switzerland: MDPI AG

Erscheinungsjahr

2020

Link zum Volltext

Quelle

EZB Free E-Journals

Beschreibungen/Notizen

• High thermostability and catalytic activity are key properties for nitrile hydratase (NHase, EC 4.2.1.84) as a well-industrialized catalyst. In this study, rational design was applied to tailor the thermostability of NHase from JCM3095 ( NHase) by combining FireProt server prediction and molecular dynamics (MD) simulation. Site-directed mutagenesis of non-catalytic residues provided by the rational design was subsequentially performed. The positive multiple-point mutant, namely, M10 (αI5P/αT18Y/αQ31L/αD92H/βA20P/βP38L/βF118W/βS130Y/βC189N/βC218V), was obtained and further analyzed. The Melting temperature ( ) of the M10 mutant showed an increase by 3.2 °C and a substantial increase in residual activity of the enzyme at elevated temperatures was also observed. Moreover, the M10 mutant also showed a 2.1-fold increase in catalytic activity compared with the wild-type NHase. Molecular docking and MD simulations demonstrated better substrate affinity and improved thermostability for the mutant.