Directed evolution of a type A feruloyl esterase for increased thermostability and organic solvent tolerance
Julien Durand, Victor Chaumeton, Olga Gherbovet, Mélanie Ragon, Peter Jütten, Joana L.A. Bras, Catarina I.P.D. Guerreiro, Carlos M.G.A. Fontes, Alexander Piechot, Régis Fauré, Sophie Bozonnet, Michael J. O’Donohue
Taros
Abstract:
Feruloyl esterases (FAE) are carboxylic esterases (EC 3.1.1) that cleave the bond between ferulic acid and L-arabinofuranosyl residues of arabinoxylans. These enzymes, in addition to helping the econstruction of lignocellulosic biomass, have a natural capacity to synthesize a wide range of bioactive molecules with interesting properties (notably anti-oxidant) in the fields of food, pharmaceuticals or cosmetics.
Within the OPTIBIOCAT European project, a methodology to express, generate and screen diversity for Aspergillus niger feruloyl esterase A was developed, tested and implemented in a high-throughput fashion, using ad hoc chromogenic probes [1]. A total of ~10,000 mutant clones were generated and screened to isolate 13 mutants with improved thermal and/or solvent stability. The most interesting mutations were individually studied while their random recombination was performed in a second round of evolution through Staggered Extension Process (StEP) [2].

Looks interesting?
As a leading contract research organization in discovery chemistry and custom synthesis, Taros provides comprehensive solutions to meet our clients’ needs from diverse branches and applications
Our experienced chemists practice a pragmatic culture and customer-centric approach in order to exceed project expectations, despite challenges, bridging very narrow timelines. They are trained to consider economic, safety and ecological boundaries and conditions laid out for all stages of the project, starting from the development of synthetic routes to…