title: Substrate structure-activity relationships guide rational engineering of modular polyketide synthase ketoreductases.
abstract: Modular polyketide synthase ketoreductases can set two chiral centers through a single reduction. To probe the basis of stereocontrol, a structure-activity relationship study was performed with three α-methyl, β-ketothioester substrates and four ketoreductases. Since interactions with the β-ketoacyl moiety were found to be most critical, residues implicated in contacting this moiety were mutated. Two mutations were sufficient to completely reverse the stereoselectivity of the model ketoreductase EryKR1, converting it from an enzyme that generates (2S,3R)-products into one that yields (2S,3S)-products.
authors.affiliation: Department of Chemistry, University of Texas at Austin, 105 E. 24th St. Stop A5300, Austin, TX 78712, USA. adriankc@utexas.edu.
authors.forename: Constance B
authors.initials: CB
authors.lastname: Bailey
authors.affiliation: Department of Chemistry, University of Texas at Austin, 105 E. 24th St. Stop A5300, Austin, TX 78712, USA. adriankc@utexas.edu.
authors.forename: Marjolein E
authors.initials: ME
authors.lastname: Pasman
authors.affiliation: Department of Chemistry, University of Texas at Austin, 105 E. 24th St. Stop A5300, Austin, TX 78712, USA. adriankc@utexas.edu and Department of Molecular Biosciences, University of Texas at Austin, 2506 Speedway Stop A5000, Austin, TX 78712, USA.
authors.forename: Adrian T
authors.initials: AT
authors.lastname: Keatinge-Clay
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013237:Stereoisomerism; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D000429:Alcohol Oxidoreductases; C087192:polyketide synthase ketoreductase