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Tailoring the speci...
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Antonopoulou, Io,1989-Luleå tekniska universitet,Kemiteknik
(författare)
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
- Artikel/kapitelEngelska2018
Förlag, utgivningsår, omfång ...
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2018-05-24
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Public Library of Science,2018
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printrdacarrier
Nummerbeteckningar
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LIBRIS-ID:oai:DiVA.org:ltu-68904
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https://urn.kb.se/resolve?urn=urn:nbn:se:ltu:diva-68904URI
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https://doi.org/10.1371/journal.pone.0198127DOI
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Språk:engelska
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Sammanfattning på:engelska
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Ämneskategori:ref swepub-contenttype
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Ämneskategori:art swepub-publicationtype
Anmärkningar
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Validerad;2018;Nivå 2;2018-05-28 (andbra)
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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.
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Biuppslag (personer, institutioner, konferenser, titlar ...)
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Hunt, CameronLuleå tekniska universitet,Kemiteknik(Swepub:ltu)camhun
(författare)
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Cerullo, GabriellaDepartment of Chemical Sciences, University of Naples "Federico II"
(författare)
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Varriale, SimonaDepartment of Chemical Sciences, University of Naples “Federico II”
(författare)
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Gerogianni, AlexandraLuleå tekniska universitet,Kemiteknik(Swepub:ltu)aleger
(författare)
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Faraco, VincenzaDepartment of Chemical Sciences, University of Naples "Federico II"
(författare)
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Rova, UlrikaLuleå tekniska universitet,Kemiteknik(Swepub:ltu)ulrok
(författare)
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Christakopoulos, PaulLuleå tekniska universitet,Kemiteknik(Swepub:ltu)pauchr
(författare)
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Luleå tekniska universitetKemiteknik
(creator_code:org_t)
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Ingår i:PLOS ONE: Public Library of Science13:51932-6203
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