New Stereoselective Biocatalysts for Carboligation and Retro-Aldol Cleavage Reactions Derived from D-Fructose 6-Phosphate Aldolase

D-Fructose 6-phosphate aldolase (FSA) catalyzes the asymmetric cross-aldol addition of phenylacetaldehyde and hydroxyacetone. We conducted structure guided saturation mutagenesis of noncatalytic active-site residues to produce new FSA variants, with the goal of widening the substrate scope of the wild-type enzyme toward a range of para- and meta-substituted arylated aldehydes. After a single generation of mutagenesis and selection, enzymes with diverse substrate selectivity scopes were identified. The kinetic parameters and stereoselectivities for a subset of enzyme/substrate combinations were determined for the reactions in both the aldol addition and cleavage reaction directions. The achieved collection of new aldolase enzymes provides new tools for controlled asymmetric synthesis of substituted aldols.

Engel, SarahWidersten (författare)
Enugala, Thilak ReddyUppsala universitet,Biokemi,Widersten(Swepub:uu)thien191 (författare)
Al-Smadi, Derar,1983-Uppsala universitet,Organisk kemi,Widersten(Swepub:uu)deral860 (författare)
Widersten, MikaelUppsala universitet,Biokemi,Widersten(Swepub:uu)mikawide (författare)
Uppsala universitetBiokemi (creator_code:org_t)

Ingår i:Biochemistry: American Chemical Society (ACS)57:40, s. 5877-58850006-29601520-4995

Av författaren/redakt...: Ma, Huan; Engel, Sarah; Enugala, Thilak ...; Al-Smadi, Derar, ...; Widersten, Mikae ...

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