Tailoring the specificity of the type C feruloyl esterase FoFaeC from Fusarium oxysporum towards methyl sinapate by rational redesign based on small molecule docking simulations

Validerad;2018;Nivå 2;2018-05-28 (andbra)
The type C feruloyl esterase FoFaeC from Fusarium oxysporum is a newly discovered enzyme with high potential for use in the hydrolysis of lignocellulosic biomass but it shows low activity towards sinapates. In this work, small molecule docking simulations were employed in order to identify important residues for the binding of the four model methyl esters of hydroxycinnamic acids, methyl ferulate/caffeate/sinapate/p-coumarate, to the predicted structure of FoFaeC. Subsequently rational redesign was applied to the enzyme’ active site in order to improve its specificity towards methyl sinapate. A double mutation (F230H/T202V) was considered to provide hydrophobic environment for stabilization of the methoxy substitution on sinapate and a larger binding pocket. Five mutant clones and the wild type were produced in Pichia pastoris and biochemically characterized. All clones showed improved activity, substrate affinity, catalytic efficiency and turnover rate compared to the wild type against methyl sinapate, with clone P13 showing a 5-fold improvement in catalytic efficiency. Although the affinity of all mutant clones was improved against the four model substrates, the catalytic efficiency and turnover rate decreased for the substrates containing a hydroxyl substitution.

Hunt, CameronLuleå tekniska universitet,Kemiteknik(Swepub:ltu)camhun (författare)
Cerullo, GabriellaDepartment of Chemical Sciences, University of Naples "Federico II" (författare)
Varriale, SimonaDepartment of Chemical Sciences, University of Naples “Federico II” (författare)
Gerogianni, AlexandraLuleå tekniska universitet,Kemiteknik(Swepub:ltu)aleger (författare)
Faraco, VincenzaDepartment of Chemical Sciences, University of Naples "Federico II" (författare)
Rova, UlrikaLuleå tekniska universitet,Kemiteknik(Swepub:ltu)ulrok (författare)
Christakopoulos, PaulLuleå tekniska universitet,Kemiteknik(Swepub:ltu)pauchr (författare)
Luleå tekniska universitetKemiteknik (creator_code:org_t)

Av författaren/redakt...: Antonopoulou, Io ...; Hunt, Cameron; Cerullo, Gabriel ...; Varriale, Simona; Gerogianni, Alex ...; Faraco, Vincenza; visa fler...; Rova, Ulrika; Christakopoulos, ...; visa färre...

Om ämnet

TEKNIK OCH TEKNOLOGIER: TEKNIK OCH TEKNO ...; och Industriell biot ...; och Bioprocessteknik

Kungliga biblioteket hanterar dina personuppgifter i enlighet med EU:s dataskyddsförordning (2018), GDPR. Läs mer om hur det funkar här.
Så här hanterar KB dina uppgifter vid användning av denna tjänst.