{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"volume":["57(32)"],"submitter":["Junker S"],"pubmed_abstract":["A structure-guided engineering of fructose-6-phosphate aldolase was performed to expand its substrate promiscuity toward aliphatic nucleophiles, that is, unsubstituted alkanones and alkanals. A \"smart\" combinatorial library was created targeting residues D6, T26, and N28, which form a binding pocket around the nucleophilic carbon atom. Double-selectivity screening was executed by high-performance TLC that allowed simultaneous determination of total activity as well as a preference for acetone versus propanal as competing nucleophiles. D6 turned out to be the key residue that enabled activity with non-hydroxylated nucleophiles. Altogether 25 single- and double-site variants (D6X and D6X/T26X) were discovered that show useful synthetic activity and a varying preference for ketone or aldehyde as the aldol nucleophiles. Remarkably, all of the novel variants had completely lost their native activity for cleavage of fructose 6-phosphate."],"journal":["Angewandte Chemie (International ed. in English)"],"pagination":["10153-10157"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC6099348"],"repository":["biostudies-literature"],"pubmed_title":["Complete Switch of Reaction Specificity of an Aldolase by Directed Evolution In Vitro: Synthesis of Generic Aliphatic Aldol Products."],"pmcid":["PMC6099348"],"pubmed_authors":["Junker S","Fessner WD","Joosten HJ","Clapes P","Roldan R"],"additional_accession":[]},"is_claimable":false,"name":"Complete Switch of Reaction Specificity of an Aldolase by Directed Evolution In Vitro: Synthesis of Generic Aliphatic Aldol Products.","description":"A structure-guided engineering of fructose-6-phosphate aldolase was performed to expand its substrate promiscuity toward aliphatic nucleophiles, that is, unsubstituted alkanones and alkanals. A \"smart\" combinatorial library was created targeting residues D6, T26, and N28, which form a binding pocket around the nucleophilic carbon atom. Double-selectivity screening was executed by high-performance TLC that allowed simultaneous determination of total activity as well as a preference for acetone versus propanal as competing nucleophiles. D6 turned out to be the key residue that enabled activity with non-hydroxylated nucleophiles. Altogether 25 single- and double-site variants (D6X and D6X/T26X) were discovered that show useful synthetic activity and a varying preference for ketone or aldehyde as the aldol nucleophiles. Remarkably, all of the novel variants had completely lost their native activity for cleavage of fructose 6-phosphate.","dates":{"release":"2018-01-01T00:00:00Z","publication":"2018 Aug","modification":"2021-02-20T06:38:06Z","creation":"2019-03-26T23:52:24Z"},"accession":"S-EPMC6099348","cross_references":{"pubmed":["29882622"],"doi":["10.1002/anie.201804831"]}}