title: Crotonase catalysis enables flexible production of functionalized prolines and carbapenams.
abstract: The biocatalytic versatility of wildtype and engineered carboxymethylproline synthases (CMPSs) is demonstrated by the preparation of functionalized 5-carboxymethylproline derivatives methylated at C-2, C-3, C-4, or C-5 of the proline ring from appropriately substituted amino acid aldehydes and malonyl-coenzyme A. Notably, compounds with a quaternary center (at C-2 or C-5) were prepared in a stereoselective fashion by engineered CMPSs. The substituted-5-carboxymethyl-prolines were converted into the corresponding bicyclic β-lactams using a carbapenam synthetase. The results demonstrate the utility of the crotonase superfamily enzymes for stereoselective biocatalysis, the amenability of carbapenem biosynthesis pathways to engineering for the production of new bicyclic β-lactam derivatives, and the potential of engineered biocatalysts for the production of quaternary centers.
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, Oxford OX1 3TA, United Kingdom.
authors.forename: Refaat B
authors.initials: RB
authors.lastname: Hamed
authors.forename: Luc
authors.initials: L
authors.lastname: Henry
authors.forename: J Ruben
authors.initials: JR
authors.lastname: Gomez-Castellanos
authors.forename: Jasmin
authors.initials: J
authors.lastname: Mecinović
authors.forename: Christian
authors.initials: C
authors.lastname: Ducho
authors.forename: John L
authors.initials: JL
authors.lastname: Sorensen
authors.forename: Timothy D W
authors.initials: TD
authors.lastname: Claridge
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Schofield
mesh_terms: D055162:Biocatalysis; D015780:Carbapenems / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D019755:Carbon-Carbon Lyases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008745:Methylation; D008958:Models, Molecular; D011392:Proline / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods*
chemical_list: D015780:Carbapenems; D011392:Proline; D019755:Carbon-Carbon Lyases; C484323:carboxymethylproline synthase