Abstract
AbstractTo design D‐amino acid dehydrogenase (DAADH) for enhanced stability, the interactions of the subunit interfaces of DAADH were analyzed. Interaction network analysis of DAADH indicated that there are only weak interactions between the A and B subunits. Several co‐evolved residue pairs were selected for mutation to enhance interfacial interactions of subunits, and 11 designed DAADHs were obtained. DA06 and DA11 were selected for experimental verification for their salt bridges are 1.4 and 1.2‐fold of that of DAwild, respectively. DA11 can maintain 93% activity in 80°C, while it was only 40% for DAwild. Thermostability study indicated the half‐life of DA11 was 2‐fold of DAwild. Molecular dynamics simulations revealed that the extraordinary stability of the DA11 was due to the formation of extra interfacial salt bridges. The article provides a strategy of mutations outside the active site of enzyme by co‐evolutionary analysis which can reduce the effect of the activity‐thermostability trade‐off.
Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
