Unifying and versatile features of flavin-dependent monooxygenases: Diverse catalysis by a common C4a-(hydro)peroxyflavin

Journal of Biological Chemistry, Journal Year: 2023, Volume and Issue: 299(12), P. 105413 - 105413

Published: Nov. 2, 2023

Flavin-dependent monooxygenases (FDMOs) are known for their remarkable versatility and crucial roles in various biological processes applications. Extensive research has been conducted to explore the structural functional relationships of FDMOs. The majority reported FDMOs utilize C4a-(hydro)peroxyflavin as a reactive intermediate incorporate an oxygen atom into wide range compounds. This review discusses analyzes recent advancements our understanding mechanistic features governing enzyme functions. State-of-the-art discoveries related common distinct properties catalytic selected discussed. Specifically, mechanisms hydroxylation, dehalogenation, halogenation, light-emitting reactions by highlighted. We also provide new analysis based on these enzymes gain insights how same can be harnessed perform variety reactions. Challenging questions obtain further breakthroughs proposed. flavin derivatives—mostly FMN or adenine dinucleotide (FAD)—which derived from vitamin B2, cofactors substrates. These exhibit versatility, they undergo one- two-electron transfers, allowing adopt multiple redox states (1Romero E. Gómez Castellanos J.R. Gadda G. Fraaije M.W. Mattevi A. Same substrate, many reactions: activation flavoenzymes.Chem. Rev. 2018; 118: 1742-1769Google Scholar, 2Toplak M. Teufel R. Three rings rule them all: versatile flavoenzymes orchestrate diversification natural products.Biochemistry. 2022; 61: 47-56Google 3Pimviriyakul P. Chaiyen Chapter one - overview flavin-dependent enzymes.in: Tamanoi F. Enzymes. Academic Press, Cambridge, MA2020: 1-36Google Scholar). ability flavins exist transient radical state (i.e. semiquinone) either through cycle (reduced substrate) light enables challenging reactivities such activation, C-C-bond formation, cleavage, C-N bond formation (4Sandoval B.A. Meichan A.J. Hyster T.K. Enantioselective hydrogen transfer: discovery promiscuity 'ene'-reductases.J. Am. Chem. Soc. 2017; 139: 11313-11316Google 5Foja Walter Jandl C. Thyrhaug Hauer J. Storch Reduced molecular single-electron reductants after photoexcitation.J. 144: 4721-4726Google 6Black M.J. Biegasiewicz K.F. Oblinsky D.G. Kudisch B. Scholes G.D. et al.Asymmetric redox-neutral cyclization catalysed 'ene'-reductases.Nat. 2020; 12: 71-75Google 7Chaiyen enigmatic reaction with oxygen.Trends Biochem. Sci. 2012; 37: 373-380Google 8Zhang Z. Feng Yang Cui H. Harrison W. Zhong D. al.Photoenzymatic enantioselective intermolecular hydroamination.Nat. Catalysis. 2023; 6: 687-694Google play pivotal diverse involved mostly metabolisms xenobiotic detoxification, biosynthetic pathways plants biosynthesis microbial secondary metabolites, neural development humans, bacteria. Non-redox galactofuranose synthesis bacteria fungi catalyzed 9Joosten V. van Berkel W.J.H. Flavoenzymes.Curr. Opin. Biol. 2007; 11: 195-202Google 10van Flavoenzymes, Chemistry of, Wiley Encyclopedia Chemical Biology. Wiley, Hoboken, NJ2008Google 11Palfey McDonald C.A. Control catalysis monooxygenases.Arch. Biophys. 2010; 493: 26-36Google 12Tanner J.J. Boechi L. Andrew McCammon Sobrado Structure, mechanism, dynamics UDP-galactopyranose mutase.Arch. 2014; 544: 128-141Google enzymes, especially (FDMOs), have drawn significant attention fields biochemistry biotechnology due involvement broad spectrum function incorporating single great potential use applications including valuable chemicals, well drug metabolisms, biodetoxification, bioremediation, biosensors (3Pimviriyakul 13Ceccoli Bianchi Rial Flavoprotein oxidative biocatalysis: recombinant expression hosts applications.Front. Microbiol. 5: 25Google 14Reis R.A.G. Li Johnson New frontiers 2021; 699108765Google comprise group that structures Based protein components, divided two main types: single-component two-component monooxygenases. Single-component bind constitutively; reduction NAD NAD(P)H substrate monooxygenation occur at active site. In contrast, require reductase generate reduced (15Chenprakhon Wongnate T. Monooxygenation aromatic compounds monooxygenases.Protein 2019; 28: 8-29Google 16Paul C.E. Eggerichs Westphal A.H. Tischler monooxygenases: biocatalysts.Biotechnol. Adv. 51107712Google is then transferred (can via diffusion) monooxygenase subsequent (17Sucharitakul Tinikul Mechanisms transfer 555-556: 33-46Google 18Sucharitakul Phongsak Entsch Svasti Ballou D.P. Kinetics p-hydroxyphenylacetate hydroxylase explain oxygenase.Biochemistry. 46: 8611-8623Google categorized eight groups (groups A-H) characteristics (16Paul 19Mascotti M.L. Juri Ayub Furnham N. Thornton J.M. Laskowski R.A. Chopping changing: evolution monooxygenases.J. Mol. 2016; 428: 3131-3146Google They activate intermediates oxygenation reactions, forming N5-peroxide oxygenating reagents (Fig. 1) (7Chaiyen 20Toplak Matthews devil details: chemical basis flavoprotein 698108732Google 21Teufel Stull F.W. Meehan Michaudel Q. Dorrestein P.C. Palfey al.Biochemical establishment characterization EncM's flavin-N5-oxide cofactor.J. 2015; 137 25: 8078-8085Google Among FDMOs, most form C4a-(hydro)peroxyflavin, which plays critical role donator nucleophilic electrophilic Herein, we focus intermediate. allows catalyze Baeyer–Villiger oxidation, sulfoxidation, epoxidation, denitration, halogenation emission 22van Kamerbeek N.M. monooxygenases, class biocatalysts.J. Biotechnol. 2006; 124: 670-689Google Understanding different control diversity would allow engineered impactful thus highlights current state-of-knowledge extensively investigated properties, particularly unique identified used explaining manifest N-hydroxylation, emission. A deep fine-tune should future designing engineering creating biocatalysts overall folding three types Rossmann fold, TIM-barrel acyl-CoA dehydrogenase fold Despite variations structures, share trait—the utilization cofactor activation. foldings arrangements facilitate recognition binding. FAD FMN; specificity predominantly determined factors (1): spatial accommodation availability space accommodate (2) binding site environment interactions within enable ADP portion molecule. Notably, this pattern distinctive characteristic 3-hydroxy-benzoate 4-hydroxylase (PHBH) Pseudomonas fluorescens halogenase (Thal) Streptomyces albogriseolus 2A) 23Entsch Massey Flavin-oxygen derivatives hydroxylation p-hydroxybenzoate hydroxylase.J. 1976; 251: 2550-2563Google 24Phintha Prakinee K. Jaruwat Lawan Visitsatthawong S. Kantiwiriyawanitch al.Dissecting low capability halogenases.J. 296100068Google 25Hanukoglu I. Proteopedia: fold: beta-alpha-beta sites.Biochem. Educ. 43: 206-209Google shared among recognizing include between polar and/or charged residues pyrophosphate ribose sugar moieties 2A). interaction prominent having mentioned above. Interestingly, possess instead FAD—such D: 4-hydroxyphenylacetate (HPA) 3-monooxygenase Escherichia coli (HpaB) (26Deng Y. Faivre Back O. Lombard Pecqueur Fontecave Structural 3-hydroxylase coli.ChemBioChem. 21: 163-170Google Scholar) dehalogenase (HadA) Ralstonia pickettii DTP0602 Scholar)—these similar those Rossman segment designated "flavin loop" 2B). loop aids ADP, enabling specifically FADH-. Conversely, (e.g., C H FDMOs) contain sites where isoalloxazine ring binds deeply structure bacterial luciferase (LuxAB) 2C). Because configuration lacks necessary binding, only FMNH- substrate. An intriguing exception observed some cases D 3-HPA Acinetobacter baumannii (HPAH, oxygenase component HPAH C2) 4-HPA (TtHpaB) Thermus thermophilus HB8, pocket situated near surface 2D) arrangement side chain extend outward structure, both FADH- utilized flavin-binding typically comprises spacious capable accommodating cofactor. exhibits combination hydrophobic regions create optimal ring. Simultaneously, finely tuned interact specific around N1, N3, C2-carbonyl oxygen, C4-carbonyl environments organized chains amino acid residue backbones. substrate-binding domain diversity, proficiently differences sequence, maintain consistent positioning above reacting C4a position re-side. area proposed takes place, facilitating generation 2, E–H) (explained more detail next section) mode commonly all occurs terminal onto appears across substrates size, polarity, steric characteristics. Examples halide ions tryptophan halogenases, hydroxylases dehalogenases, cyclohexyl ketone cyclohexanone (CHMO) (27Blasiak L.C. Drennan C.L. perspective enzymatic halogenation.Acc. Res. 2009; 42: 147-155Google 28Agarwal Miles Z.D. Winter Eustáquio A.S. El Gamal A.A. Moore B.S. Enzymatic dehalogenation pervasive mechanistically diverse.Chem. 117: 5619-5674Google 29Yachnin B.J. Sprules McEvoy M.B. Lau P.C.K. Berghuis A.M. substrate-bound crystal baeyer–villiger criegee-like conformation.J. 134: 7788-7795Google 30Alfieri Fersini Ruangchan Prongjit Structure monooxygenase.Proc. Natl. Acad. U. 104: 1177-1182Google 31Pimviriyakul Chitnumsub HadA inhibition quadruple π-stacking.J. 297100952Google likely facilitates robust convenient It noted re-side C4a-(hydro)peroxyflavin-forming not N5-oxide. case N5-oxide, was found si-face flavin, while approach (32Matthews Saleem-Batcha Sanders J.N. Houk K.N. Aminoperoxide adducts expand repertoire monooxygenases.Nat. 16: 556-563Google explained results computational calculations demonstrated mechanism N5-oxide brought closer N5-inversion With greater information regarding it interesting conduct comparative investigations broader utilizations. may deeper controlling lead modify As described above, arrangement, positioned fundamental question arises: does buried secluded solvent react intermediate? shed process. diffuses tunnels, reaching oxygen-binding located ring, efficient intermediate, shields surrounding environment, its prompt Recent stabilizing enterocin (flavin monooxygenase) maritimus (33Saleem-Batcha al.Enzymatic dioxygen functionalization cofactor.Proc. 115: 4909-4914Google pyrimidine (RutA) location 2I). data confirming other 2J) (30Alfieri 34Visitsatthawong Chenprakhon Surawatanawong Mechanism monooxygenase: nearly barrierless C4a-hydroperoxyflavin proton-coupled electron transfer.J. 137: 9363-9374Google significantly advance demonstrate effectively order essential functionalization. feature during stability varies enzymes. cases, detected using rapid kinetic studies (35Yeh Blasiak Koglin Walsh C.T. Chlorination long-lived halogenases.Biochemistry. 1284-1292Google 36Ruangchan Tongsook Sucharitakul pH-dependent reveal 3-hydroxylase.J. 2011; 286: 223-233Google stabilize key distinctions flavoenzyme oxidases could cycles, because formed rapidly decays (37Massey Activation flavoproteins.J. 1994; 269: 22459-22462Google 38Mattevi To oxidase: reactivity flavoenzymes.Trends 31: 276-283Google 39Chakraborty Ortiz-Maldonado Studies aeruginosa: system composed small large 49: 372-385Google exceptions were pyranose 2-oxidase Trametes multicolor, half-reaction, choline oxidase C4a-adduct single-crystal spectroscopic method crystallization (40Sucharitakul Haltrich Detection oxidase.Biochemistry. 2008; 47: 8485-8490Google 41Orville Lountos G.T. Finnegan Prabhakar Crystallographic, spectroscopic, C4a−Oxygen adduct 48: 720-728Google Comparison instrumental identifying C4a-N5 locus Site-directed mutagenesis significance Ser171, close proximity to, H-bond N5, stabilization 2K). Replacing Ala resulted dramatic decrease (42Thotsaporn Stabilization achieved N5 atoms.J. 28170-28180Google Similar Thr TtHpaB (43Kim S.H. Hisano Takeda Iwasaki Ebihara Miki Crystal HB8.J. 282: 33107-33117Google Scholar), chlorophenol 4-monooxygenase (TftD) Burkholderia cepacia AC1100 (44Webb B.N. Ballinger J.W. Kim Belchik S.M. Lam K.-S. Youn al.Characterization NADH:FAD oxidoreductase (TftC) AC1100.J. 285: 2014-2027Google (31Pimviriyakul cycles 45Pimviriyakul Thotsaporn Kinetic dechlorinating HadA.J. 292: 4818-4832Google H-bonding thought important stabilization. (BVMOs) N-hydroxylating (NMOs), despite absence NADP+ required (for periods up hours, is, CHMO sp. NCIMB 9871 (46Sheng Mechanistic monooxy

Language: Английский

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Expanding the toolbox of Baeyer–Villiger and flavin monooxygenase biocatalysts for the enantiodivergent green synthesis of sulfoxides

Green Chemistry, Journal Year: 2024, Volume and Issue: 26(15), P. 8685 - 8693

Published: Jan. 1, 2024

Two new monooxygenase biocatalysts, the Baeyer-Villiger BVMO145 and flavin FMO401 from Almac library, have been found to catalyse enantiodivergent oxidation of sulfides bearing N-heterocyclic substituents into sulfoxides under mild green conditions. The biocatalyst provides (

Language: Английский

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Hydroxyl Radical Mediated Heterogeneous Photocatalytic Baeyer‐Villiger Oxidation over Covalent Triazine/Heptazine‐Based Frameworks

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 18, 2024

The Baeyer-Villiger (B-V) oxidation of ketones to the corresponding lactones/esters is a classic and essential reaction in chemical industry. However, this process has not yet been achieved ambient conditions with aid oxygen heterogeneous photocatalysts. In study, we developed an organic photocatalytic system using covalent triazine/heptazine-based frameworks (CTF-TB/CHF-TB) enable B-V under mild through cascade pathway. Experimental data theoretical calculations showed that heptazine/triazine units can "chelate" decompose situ generated H

Language: Английский

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Evolution of Glucose Dehydrogenase for Cofactor Regeneration in Bioredox Processes with Denaturing Agents

ChemBioChem, Journal Year: 2020, Volume and Issue: 21(18), P. 2680 - 2688

Published: April 23, 2020

Glucose dehydrogenase (GDH) is a general tool for driving nicotinamide (NAD(P)H) regeneration in synthetic biochemistry. An increasing number of bioreactions are carried out media containing high amounts organic cosolvents or hydrophobic substrates/products, which often denature native enzymes, including those cofactor regeneration. In this work, we attempted to improve the chemical stability Bacillus megaterium GDH (BmGDHM0 ) presence large 1-phenylethanol by directed evolution. Among resulting mutants, BmGDHM6 (Q252L/E170K/S100P/K166R/V72I/K137R) exhibited 9.2-fold increase tolerance against 10 % (v/v) 1-phenylethanol. Moreover, was also more stable than BmGDHM0 when exposed and enzyme-inactivating compounds such as acetophenone, ethyl 2-oxo-4-phenylbutyrate, (R)-2-hydroxy-4-phenylbutyrate. Coupled with Candida glabrata carbonyl reductase, successfully used asymmetric reduction deactivating 2-oxo-4-phenylbutyrate total turnover 1800 cofactor, thus making it attractive commercial application. Overall, evolution chemically robust facilitates its wider use NAD(P)H biocatalysis.

Language: Английский

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The Unexplored Importance of Fleeting Chiral Intermediates in Enzyme-Catalyzed Reactions

Journal of the American Chemical Society, Journal Year: 2021, Volume and Issue: 143(37), P. 14939 - 14950

Published: Sept. 7, 2021

Decades of extensive research efforts by biochemists, organic chemists, and protein engineers have led to an understanding the basic mechanisms essentially all known types enzymes, but in a formidable number cases essential aspect has been overlooked. The occurrence short-lived chiral intermediates formed symmetry-breaking prochiral precursors enzyme catalyzed reactions systematically neglected. We designate these elusive species as fleeting analyze such crucial questions "Do occur homochiral form?" If so, what is absolute configuration, why did Nature choose that particular stereoisomeric form, even when isolable final product may be achiral? Does configuration depend any way on precursor? How does this affect catalytic proficiency enzyme? issues continue unexplored, then many remains incomplete. systematized according their structures types. This followed critical analyses selected case studies conclusions perspectives. hope fascinating concept will attract attention scientists, thereby opening exciting new field.

Language: Английский

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