title: Characterizing and predicting carboxylic acid reductase activity for diversifying bioaldehyde production.
abstract: Chemicals with aldehyde moieties are useful in the synthesis of polymerization reagents, pharmaceuticals, pesticides, flavors, and fragrances because of their high reactivity. However, chemical synthesis of aldehydes from carboxylic acids has unfavorable thermodynamics and limited specificity. Enzymatically catalyzed reductive bioaldehyde synthesis is an attractive route that overcomes unfavorable thermodynamics by ATP hydrolysis in ambient, aqueous conditions. Carboxylic acid reductases (Cars) are particularly attractive, as only one enzyme is required. We sought to increase the knowledge base of permitted substrates for four Cars. Additionally, the Lys2 enzyme family was found to be mechanistically the same as Cars and two isozymes were also tested. Our results show that Cars prefer molecules where the carboxylic acid is the only polar/charged group. Using this data and other published data, we develop a support vector classifier (SVC) for predicting Car reactivity and make predictions on all carboxylic acid metabolites in iAF1260 and Model SEED.
authors.affiliation: Department of Chemical and Biological Engineering, Northwestern University, Evanston, 60208, Illinois.
authors.forename: Matthew
authors.initials: M
authors.lastname: Moura
authors.affiliation: Department of Chemical and Biological Engineering, Northwestern University, Evanston, 60208, Illinois.
authors.forename: Dante
authors.initials: D
authors.lastname: Pertusi
authors.affiliation: Department of Chemical and Biological Engineering, Northwestern University, Evanston, 60208, Illinois.
authors.forename: Stephen
authors.initials: S
authors.lastname: Lenzini
authors.affiliation: Department of Chemical and Biological Engineering, Northwestern University, Evanston, 60208, Illinois.
authors.forename: Namita
authors.initials: N
authors.lastname: Bhan
authors.affiliation: Department of Chemical and Biological Engineering, Northwestern University, Evanston, 60208, Illinois. broadbelt@northwestern.edu.
authors.forename: Linda J
authors.initials: LJ
authors.lastname: Broadbelt
authors.affiliation: Department of Chemical and Biological Engineering, Northwestern University, Evanston, 60208, Illinois. k-tyo@northwestern.edu.
authors.forename: Keith E J
authors.initials: KE
authors.lastname: Tyo
mesh_terms: D000447:Aldehydes / Q000378:metabolism*; D002264:Carboxylic Acids / Q000378:metabolism*; D003198:Computer Simulation; D007218:Industrial Microbiology / Q000379:methods; D008954:Models, Biological; D009161:Mycobacterium / Q000201:enzymology*; D009249:NADP / Q000378:metabolism; D009615:Nocardia / Q000201:enzymology*; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism*; D013379:Substrate Specificity; D060388:Support Vector Machine; D013816:Thermodynamics
chemical_list: D000447:Aldehydes; D002264:Carboxylic Acids; D009249:NADP; D010088:Oxidoreductases; C019825:carboxylic acid reductase
keywords: carboxylic acid reductase; enzyme promiscuity; reductive aldehyde synthesis; support vector machines