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The Role of Substra...
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Enugala, Thilak ReddyUppsala universitet,Biokemi,Widersten
(author)
The Role of Substrate-Coenzyme Crosstalk in Determining Turnover Rates in Rhodococcus ruber Alcohol Dehydrogenase
- Article/chapterEnglish2020
Publisher, publication year, extent ...
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2020-07-16
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American Chemical Society (ACS),2020
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electronicrdacarrier
Numbers
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LIBRIS-ID:oai:DiVA.org:uu-416371
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https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-416371URI
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https://doi.org/10.1021/acscatal.0c01654DOI
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Language:English
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Summary in:English
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Subject category:ref swepub-contenttype
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Subject category:art swepub-publicationtype
Notes
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Eight related alcohol dehydrogenases that had been originally isolated by laboratory evolution of ADH-A from Rhodococcus ruber DSM44541 for modified substrate scopes, were together with their parent wild-type, subjected to biochemical characterization of possible activities with a panel of chiral alcohols and pro-chiral ketones. Determinations of rates of catalyzed alcohol oxidations and ketone reductions, and of cofactor release, pointed out to the role of a W295A substitution as being decisive in steering enantioselectivity in the oxidation of arylated 1-methyl substituted alcohols. Molecular dynamics simulations of enzyme-substrate interactions in the Michaelis complexes of wild-type and a Y294F/W295A double mutant could rationalize the experimentally observed shift in enantioselectivity and differences in catalytic activity with 4-phenyl-2-butanol. Finally, we present herein evidence for apparent inter-dependency between substrate/product and the cofactor in the ternary complex, that directly affects the NADH dissociation rates, and thus that this substrate-coenzyme crosstalk plays a direct role in determining the turnover rates.
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Corbella Morató, MarinaUppsala universitet,Biokemi,Kamerlin(Swepub:uu)marco394
(author)
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Kamerlin, Shina C. L.,1981-Uppsala universitet,Biokemi,Kamerlin(Swepub:uu)lynka392
(author)
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Widersten, MikaelUppsala universitet,Biokemi,Widersten(Swepub:uu)mikawide
(author)
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Uppsala universitetBiokemi
(creator_code:org_t)
Related titles
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In:ACS Catalysis: American Chemical Society (ACS)10:16, s. 9115-91282155-5435
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