An effective computational-screening strategy for simultaneously
improving both catalytic activity and thermostability of
α-L-rhamnosidase
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.