Evidence for the participation of an extra α-helix at β-subunit surface in the thermal stability of Co-type nitrile hydratase

Applied Microbiology and Biotechnology
Xiaolin PeiAnming Wang

Abstract

Nitrile hydratase (NHase) has attracted considerable attention since it can efficiently catalyze the hydration of nitriles to valuable amides. However, the poor stability of NHase is one of the main drawbacks in the industrial application. In this study, we compared the structural difference between Fe-type and Co-type NHase and found that an extra α helix existed at the β-subunit surface of Co-type NHase (defined as the β-6th helix). Then, the effects of the β-6th helix were investigated on the thermal stability and the catalytic kinetics of a Co-type NHase from Aurantimonas manganoxydans ATCC BAA-1229 (NHase1229). When the β-6th helix was deleted or disrupted, the thermal stability of NHase1229 was reduced to 17.6 and 12.9% of that of wild NHase1229, respectively. Thus, the β-6th helix is important for the thermal stability of Co-type NHase. Based on the structural characteristics of Co-type NHase, the β-6th helix may be interacted with another helix at the α-subunit (defined as the α-2nd helix) by hydrophobic network just as a "magnetic suction buckle" on the enzyme surface to stabilize the binding of α- and β-subunits. The β-6th helix is located at the mouth of the substrate and product tunnel, so it plays crucial roles in ...Continue Reading

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Citations

Dec 21, 2019·Applied Microbiology and Biotechnology·Song JiaoZhongyao Shen
May 12, 2020·Frontiers in Bioengineering and Biotechnology·Zhongyi ChengZhemin Zhou
Oct 23, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Zhongyi ChengLukasz Peplowski
Sep 2, 2021·Journal of Agricultural and Food Chemistry·Shilei SunMiaomiao Sheng

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Datasets Mentioned

BETA
BAA-1229
KP236109
JCM3095
HV233495

Methods Mentioned

BETA
PCR
electrophoresis
Protein Assay
circular dichroism

Software Mentioned

PyMOL
Porter
Accelry Discovery Studio
Clustal Omega
PROCHECK
SCRATCH

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