Ancestral Resurrection and Directed Evolution of Fungal Mesozoic Laccases
Authors/Creators
- 1. aDepartment of Biocatalysis, Institute of Catalysis and Petrochemistry, CSIC, Madrid, Spain, bEvoEnzyme, S.L., Madrid, Spain cDepartamento de Química Física, Facultad de Ciencias, Universidad de Granada, Granada, Spain dINRS–Institut Armand-Frappier, Université du Québec, Laval, Quebec, Canada
Description
Ancestral sequence reconstruction and resurrection provides useful information
for protein engineering, yet its alliance with directed evolution has been
little explored. In this study, we have resurrected several ancestral nodes of fungal
laccases dating back !500 to 250 million years. Unlike modern laccases, the resurrected
Mesozoic laccases were readily secreted by yeast, with similar kinetic parameters,
a broader stability, and distinct pH activity profiles. The resurrected Agaricomycetes
laccase carried 136 ancestral mutations, a molecular testimony to its origin,
and it was subjected to directed evolution in order to improve the rate of 1,3-
cyclopentanedione oxidation, a !–diketone initiator commonly used in vinyl polymerization
reactions.
Files
2020-Applied&EnvirnomentalMicrobiology.pdf
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