Reaction coordinate of isopenicillin N synthase: oxidase versus oxygenase activity.
Isopenicillin N synthase (IPNS) can have both oxidase and oxygenase activity depending on the substrate. For the native substrate, ACV, oxidase activity exists; however, for the substrate analogue ACOV, which lacks an amide nitrogen, IPNS exhibits oxygenase activity. The potential energy surfaces for the O-O bond elongation and cleavage were calculated for three different reactions: homolytic cleavage via traditional Fenton chemistry, heterolytic cleavage, and nucleophilic attack. These surfaces show that the hydroperoxide-ferrous intermediate, formed by O(2)-activated H atom abstraction from the substrate, can exploit different reaction pathways and that interactions with the substrate govern the pathway. The hydrogen bonds from hydroperoxide to the amide nitrogen of ACV polarize the sigma* orbital of the peroxide toward the proximal oxygen, facilitating heterolytic cleavage. For the substrate analogue ACOV, this hydrogen bond is no longer present, leading to nucleophilic attack on the substrate intermediate C-S bond. After cleavage of the hydroperoxide, the two reaction pathways proceed with minimal barriers, resulting in the closure of the beta-lactam ring for the oxidase activity (ACV) or formation of the thiocarboxylate for oxygenase activity (ACOV).
Brown-Marshall
Christina D
CD
Department of Chemistry, Stanford University, Stanford, California 94305, USA.
Diebold
Adrienne R
AR
Solomon
Edward I
EI
D001233:Aspergillus nidulans / Q000201:enzymology*; D004579:Electron Transport; D004789:Enzyme Activation; D005296:Ferrous Compounds / Q000737:chemistry / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry* / Q000378:metabolism; D006860:Hydrogen Bonding; D006868:Hydrolysis; D010088:Oxidoreductases / Q000737:chemistry* / Q000378:metabolism; D010105:Oxygenases / Q000737:chemistry* / Q000378:metabolism; D010406:Penicillins; D010453:Peptide Synthases / Q000737:chemistry / Q000378:metabolism; D010545:Peroxides / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity; D013816:Thermodynamics
D005296:Ferrous Compounds; D005656:Fungal Proteins; D010406:Penicillins; D010545:Peroxides; D010088:Oxidoreductases; D010105:Oxygenases; C038370:isopenicillin N synthetase; D010453:Peptide Synthases; C095368:alpha-aminoadipyl-cysteinyl-valine synthetase; C006414:penicillin N