Insight into the broadened substrate scope of nitrile hydratase by static and dynamic structure analysis

• DOI: 10.1039/d2sc02319a
• Publication Date: 2022-07-06T08:45:30Z
• Authors: Dong Ma; Zhongyi Cheng; Lukasz Peplowski; Laichuang Han; Yuanyuan Xia; Xiaodong Hou; Junling Guo; Dejing Yin; Yijian Rao; Zhemin Zhou
• Keywords: Scope (computer science); Nitrile hydratase

The narrow substrate scope limits the wide industrial application of enzymes. Here, we successfully broadened a nitrile hydratase (NHase) through mutation two tunnel entrance residues based on rational calculation. Two variants, with increased specific activity, especially toward bulky substrates, were obtained. Crystal structure analysis revealed that mutations led to expansion entrance, which might be conducive entry. More importantly, molecular dynamics simulations illustrated introduced anti-correlated movements regions around and active site, would promote access during dynamic process catalysis. Additionally, corresponding other NHases also enhanced their activity substrates. These results not only located at enzyme surface key factor in catalytic performance, but provided evidence for insight into broadening.