title: Truncated FAD synthetase for direct biocatalytic conversion of riboflavin and analogs to their corresponding flavin mononucleotides.
abstract: The preparation of flavin mononucleotide (FMN) and FMN analogs from their corresponding riboflavin precursors is traditionally performed in a two-step procedure. After initial enzymatic conversion of riboflavin to flavin adenine dinucleotide (FAD) by a bifunctional FAD synthetase, the adenyl moiety of FAD is hydrolyzed with snake venom phosphodiesterase to yield FMN. To simplify the protocol, we have engineered the FAD synthetase from Corynebacterium ammoniagenes by deleting its N-terminal adenylation domain. The newly created biocatalyst is stable and efficient for direct and quantitative phosphorylation of riboflavin and riboflavin analogs to their corresponding FMN cofactors at preparative-scale.
authors.affiliation: Department of Chemistry, Emory University, 1515 Dickey Drive, Atlanta, GA 30322, USA.
authors.forename: Samantha M
authors.initials: SM
authors.lastname: Iamurri
authors.forename: Ashley B
authors.initials: AB
authors.lastname: Daugherty
authors.forename: Dale E
authors.initials: DE
authors.lastname: Edmondson
authors.forename: Stefan
authors.initials: S
authors.lastname: Lutz
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003352:Corynebacterium / Q000201:enzymology / Q000235:genetics; D005486:Flavin Mononucleotide / Q000378:metabolism*; D008958:Models, Molecular; D009713:Nucleotidyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012256:Riboflavin / Q000031:analogs & derivatives / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D005486:Flavin Mononucleotide; D009713:Nucleotidyltransferases; C020143:FMN adenylyltransferase; D012256:Riboflavin
keywords: FAD synthetase; biocatalysis; cofactor analogs; protein engineering