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An effective computational-screening strategy for simultaneously improving both catalytic activity and thermostability of α-L-rhamnosidase
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  • Lijun Li,
  • Wenjing Li,
  • Jianye Gong,
  • Yanyan Xu,
  • Zheyu Wu,
  • Zedong Jiang,
  • Yi-sheng Cheng,
  • Qingbiao Li,
  • Hui Ni
Lijun Li
Jimei University

Corresponding Author:[email protected]

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Wenjing Li
Jimei University
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Jianye Gong
Jimei University
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Yanyan Xu
Xiamen University
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Zheyu Wu
Jimei University
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Zedong Jiang
Jimei University
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Yi-sheng Cheng
National Taiwan University
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Qingbiao Li
Xiamen University
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Hui Ni
Jimei University
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Abstract

Catalytic efficiency and thermostability are the two most important characteristics of enzymes. However, it is always tough to improve both catalytic efficiency and thermostability of enzymes simultaneously. In the present study, a computational strategy with double-screening steps was proposed to simultaneously improve both catalysis efficiency and thermostability of enzymes; and a fungal α-L-rhamnosidase was used to validate the strategy. As the result, by molecular docking and sequence alignment analysis within the binding pocket, seven mutant candidates were predicted with better catalytic efficiency. By energy variety analysis, three among the seven mutant candidates were predict with better thermostability. The expression and characterization results showed the mutant D525N had significant improvements in both enzyme activity and thermostability. Molecular dynamics simulations indicated that the mutations located within the 5 Å range of the catalytic domain, which could improve RMSD, electrostatic, Van der Waal interaction and polar salvation values, and formed water bridge between the substrate and the enzyme. The study indicated that the computational strategy based on the binding energy, conservation degree and mutation energy analyses was effective to develop enzymes with better catalysis and thermostability, providing practical approach for developing industrial enzymes.
27 Jan 2021Submitted to Biotechnology and Bioengineering
27 Jan 2021Submission Checks Completed
27 Jan 2021Assigned to Editor
28 Jan 2021Reviewer(s) Assigned
09 Feb 2021Editorial Decision: Revise Minor
09 Feb 2021Review(s) Completed, Editorial Evaluation Pending
04 Mar 20211st Revision Received
04 Mar 2021Submission Checks Completed
04 Mar 2021Assigned to Editor
12 Mar 2021Reviewer(s) Assigned
18 Mar 2021Review(s) Completed, Editorial Evaluation Pending
18 Mar 2021Editorial Decision: Accept