@misc{10481/62664,
year = {2020},
month = {5},
url = {http://hdl.handle.net/10481/62664},
abstract = {Among the broad repertory of protein engineering methods that set out to improve
stability, consensus design has proved to be a powerful strategy to stabilize enzymes
without compromising their catalytic activity. Here, we have applied an in-house
consensus method to stabilize a laboratory evolved high-redox potential laccase.
Multiple sequence alignments were carried out and computationally refined by applying
relative entropy and mutual information thresholds. Through this approach, an ensemble
of 20 consensus mutations were identified, 18 of which were consensus/ancestral
mutations. The set of consensus variants was produced in Saccharomyces cerevisiae
and analyzed individually, while site directed recombination of the best mutations did
not produce positive epistasis. The best single variant carried the consensus-ancestral
A240G mutation in the neighborhood of the T2/T3 copper cluster, which dramatically
improved thermostability, kinetic parameters and secretion.},
organization = {This study is based upon work funded by and the Spanish
Government projects BIO2013-43407-R-DEWRY and BIO2016-
79106-R-Lignolution. BG-F was supported by a FPI national
fellowship BES-2014-068887.},
publisher = {Frontiers Media},
keywords = {Consensus design},
keywords = {High-redox potential laccase},
keywords = {Ancestor mutation},
keywords = {Thermostability},
keywords = {Activity},
title = {Consensus Design of an Evolved High-Redox Potential Laccase},
doi = {10.3389/fbioe.2020.00354},
author = {Gomez-Fernandez, Bernardo J. and Risso, Valeria Alejandra and Sánchez Ruiz, José Manuel},
}