How pH Modulates the Reactivity and Selectivity of a Siderophore-Associated Flavin Monooxygenase
نویسندگان
چکیده
Flavin-containing monooxygenases (FMOs) catalyze the oxygenation of diverse organic molecules using O2, NADPH, and the flavin adenine dinucleotide (FAD) cofactor. The fungal FMO SidA initiates peptidic siderophore biosynthesis via the highly selective hydroxylation of L-ornithine, while the related amino acid L-lysine is a potent effector of reaction uncoupling to generate H2O2. We hypothesized that protonation states could critically influence both substrate-selective hydroxylation and H2O2 release, and therefore undertook a study of SidA's pH-dependent reaction kinetics. Consistent with other FMOs that stabilize a C4a-OO(H) intermediate, SidA's reductive half reaction is pH independent. The rate constant for the formation of the reactive C4a-OO(H) intermediate from reduced SidA and O2 is likewise independent of pH. However, the rate constants for C4a-OO(H) reactions, either to eliminate H2O2 or to hydroxylate L-Orn, were strongly pH-dependent and influenced by the nature of the bound amino acid. Solvent kinetic isotope effects of 6.6 ± 0.3 and 1.9 ± 0.2 were measured for the C4a-OOH/H2O2 conversion in the presence and absence of L-Lys, respectively. A model is proposed in which L-Lys accelerates H2O2 release via an acid-base mechanism and where side-chain position determines whether H2O2 or the hydroxylation product is observed.
منابع مشابه
Effect of NaNO2, HNO3 and H2SO4 on the structure and reactivity of gamma-alumina
The influence of NaNO2, HNO3 and H2SO4 on the structure and morphology of gamma-alumina was investigated by XRD, BET, SEM and FT-IR spectroscopy. Selected amounts of NaNO2, HNO3 and H2SO4 were added to a solution of aluminum isopropoxide (as a precursor for the boehmite synthesis). The boehmite samples were calcined at 350 and 600 ℃ to form the semi-crystalline high surface area gamma-Al2O3. Th...
متن کاملCharacterization of the Ornithine Hydroxylation Step in Albachelin Biosynthesis.
N-Hydroxylating monooxygenases (NMOs) are involved in siderophore biosynthesis. Siderophores are high affinity iron chelators composed of catechol and hydroxamate functional groups that are synthesized and secreted by microorganisms and plants. Recently, a new siderophore named albachelin was isolated from a culture of Amycolatopsis alba growing under iron-limiting conditions. This work focuses...
متن کاملاثر سیدروفور دسفرال (DFOB) بر جذب کادمیم بهوسیله کانی مسکویت
Siderophores are low molecular weight iron-binding ligands produced by aerobic soil microorganisms and some plants roots. They may also develop complexes with other metals such as Cd, thereby influencing the bio-availability, toxicity and mobility of Cd in the soils and aqueous environments. The main objective of this study was to investigate the effects of desferal siderophore (DFOB) on the ad...
متن کاملNoncanonical Reactions of Flavoenzymes
Enzymes containing flavin cofactors are predominantly involved in redox reactions in numerous cellular processes where the protein environment modulates the chemical reactivity of the flavin to either transfer one or two electrons. Some flavoenzymes catalyze reactions with no net redox change. In these reactions, the protein environment modulates the reactivity of the flavin to perform novel ch...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
دوره 53 شماره
صفحات -
تاریخ انتشار 2014