A5073588774- Pharmacogenetics and Drug Metabolism
- Metal-Catalyzed Oxygenation Mechanisms
- Porphyrin and Phthalocyanine Chemistry
- Hemoglobin structure and function
- Cancer Treatment and Pharmacology
- Porphyrin Metabolism and Disorders
- Computational Drug Discovery Methods
- Eicosanoids and Hypertension Pharmacology
- Heme Oxygenase-1 and Carbon Monoxide
- Enzyme Catalysis and Immobilization
- Analytical Chemistry and Chromatography
- Asymmetric Hydrogenation and Catalysis
- Chemical Reactions and Isotopes
- Microbial Metabolic Engineering and Bioproduction
- DNA and Nucleic Acid Chemistry
- Crystallization and Solubility Studies
- X-ray Diffraction in Crystallography
- RNA Interference and Gene Delivery
- Synthesis and Catalytic Reactions
- Advanced biosensing and bioanalysis techniques
- Oxidative Organic Chemistry Reactions
- Cancer, Hypoxia, and Metabolism
- Mass Spectrometry Techniques and Applications
- Metal complexes synthesis and properties
- Chemical Synthesis and Analysis
Nagoya University
2016-2025
Japan Science and Technology Agency
2016-2023
Graduate School USA
2015-2020
Centre for Research in Engineering Surface Technology
2017
Science and Technology Corporation (United States)
2017
Centre de Recherche en Économie et Statistique
2017
Ewha Womans University
2009
Nara Institute of Science and Technology
2005-2006
Osaka University
2005
Chiba University
2001-2003
Playing tricks on enzymes: Direct hydroxylation of benzene to phenol was catalyzed by wild-type P450BM3 in the presence perfluorinated carboxylic acids as decoy molecules. The catalytic turnover rate reached 120 min−1 per P450. selectivity towards production very high and no overoxidation products were detected. Phenol is a key intermediate industry for synthesis drugs, dyes, functional polymers. Because currently produced cumene process,1 which involves energy consumption significant...
It has long been believed that the fatty acid hydroxylase wild-type P450BM3 is unable to oxidize gaseous alkanes. However, simple addition of a perfluorocarboxylic as dummy substrate initiate catalytic cycle enabled efficient hydroxylation butane and propane (see picture).
Cytochrome P450(SPα) (CYP152B1) isolated from Sphingomonas paucimobilis is the first P450 to be classified as a H(2)O(2)-dependent P450. hydroxylates fatty acids with high α-regioselectivity. Herein we report crystal structure of palmitic acid substrate at resolution 1.65 Å. The revealed that C(α) bound in one alternative conformations 4.5 Å heme iron. This conformation explains highly selective α-hydroxylation observed P450(SPα). Mutations active site and F-G loop did not impair its...
Time-resolved serial femtosecond crystallography using an X-ray free electron laser (XFEL) in conjunction with a photosensitive caged-compound offers crystallographic method to track enzymatic reactions. Here we demonstrate the application of this fungal NO reductase, heme-containing enzyme, at room temperature. Twenty milliseconds after caged-NO photolysis, identify NO-bound form which is initial intermediate slightly bent Fe-N-O coordination geometry resolution 2.1 Å. The compatible those...
Abstract We report a unique strategy for the development of H 2 O ‐dependent cytochrome P450BM3 system, which catalyzes monooxygenation non‐native substrates with assistance dual‐functional small molecules (DFSMs), such as N ‐(ω‐imidazolyl fatty acyl)‐ l ‐amino acids. The acyl amino acid group DFSM is responsible bounding to enzyme an anchoring group, while imidazolyl plays role general acid–base catalyst in activation . This system affords best peroxygenase activity epoxidation styrene,...
Trick or treat: Cytochrome P450BSβ was transformed into a monooxygenase suitable for practical use by employing simple substrate trick. The specificity of altered drastically decoy molecule, while its intrinsic advantage, the hydrogen peroxide, retained. catalytic activities and enantioselectivity H2O2–P450BSβ system are highly dependent on structure molecule. Supporting information this article is available WWW under http://www.wiley-vch.de/contents/jc_2002/2007/z700068_s.pdf from author....
Ferrocene-bridged trisporphyrin (2) was synthesized by two-steps condensation of corresponding aldehydes and dipyrromethanes, its self-assembling behavior based on the complementary coordination motif imidazolylporphyrinatozinc(II) investigated in conjunction with hinge-like flexibility given freely rotating cyclopentadienyl rings ferrocene connector. spontaneously exclusively generated dimeric ring (7) upon simple zinc(II) insertion, indicating that hinge connector favored smallest...
The direct hydroxylation of alkanes under mild conditions is a key issue in catalytic chemistry that addresses an increasing number industrial and economic requirements. Cytochrome P450s are monooxygenases capable oxidizing less reactive C–H bonds; however, wild-type unavailable for many important nonnative substrates such as gaseous alkanes. Here, we report the enhanced activities crystallographic evidence role decoy molecules P450BM3-catalyzed ethane propane by using next generation...
The selective hydroxylation of benzene to phenol, without the formation side products resulting from overoxidation, is catalyzed by cytochrome P450BM3 with assistance amino acid derivatives as decoy molecules. catalytic turnover rate and total number reached 259 min-1 P450BM3-1 40 200 when N-heptyl-l-proline modified l-phenylalanine (C7-l-Pro-l-Phe) was used molecule. This work shows that a totally different structure fatty acids can be molecules for aromatic wild-type P450BM3. method...
Abstract Nuclear import receptors (NIRs) not only transport RNA-binding proteins (RBPs) but also modify phase transitions of RBPs by recognizing nuclear localization signals (NLSs). Toxic arginine-rich poly-dipeptides from C9orf72 interact with NIRs and cause nucleocytoplasmic deficit. However, the molecular basis for toxicity toward function as modifiers remains unidentified. Here we show that impede ability to RBPs. Isothermal titration calorimetry size-exclusion chromatography revealed...
Coordination-assembled porphyrin macrocycles reinforced with covalent bondings were deposited on a metal surface by pulse injection method, and their scanning tunneling microscopy (STM) images recorded under ultrahigh vacuum conditions at liquid nitrogen temperature. The decamer ring consisting of 30 porphyrins gave clear circular STM hollow structure, whereas that without linking did not give images, showing the coordination pairs ring-closing metathesis reaction was effective to reinforce...
H<sub>2</sub>O<sub>2</sub>-dependent cytochrome P450s that can catalyze monooxygenation of nonnative substrates were constructed by one-point mutagenesis.
Cytochrome P450BM3 (P450BM3) is a long-alkyl-chain fatty acid hydroxylase that shows an extremely high catalytic turnover rate and coupling efficiency of NADPH for product formation (product per consumption rate). Although exclusively hydroxylates acids, we have found simple addition perfluorocarboxylic acids (PFs) as inert dummy substrates (decoy molecules) turns into small alkane hydroxylase. For example, PF-bound oxidizes propane butane to 2-propanol 2-butanol, respectively. The...
An Escherichia coli whole-cell biocatalyst for the direct hydroxylation of benzene to phenol has been developed. By adding amino acid derivatives as decoy molecules culture medium, wild-type cytochrome P450BM3 (P450BM3) expressed in E.coli can be activated and non-native substrates hydroxylated, without supplementing with NADPH. The yield reached 59 % when N-heptyl-l-prolyl-l-phenylalanine (C7-Pro-Phe) was employed molecule. It shown that molecules, especially those lacking fluorination,...
ConspectusBacterial cytochrome P450s (P450s) are at the focus of attention as potential biocatalysts for applications in green synthetic chemistry, they possess high activity hydroxylation inert substrate C–H bonds. The bacterial P450s, such P450BM3, is chiefly due to their specificity, and consequently, catalytic P450BM3 toward non-native substrates very low, limiting utility biocatalysts. To enable oxidation by without any mutagenesis, we have developed a series "decoy molecules", dummy...
Biological methane oxidation is a highly desirable method for the conversion of natural gas into liquid to meet increasing demand fuel and chemical feedstock as well reducing potent greenhouse effects emissions. Because hemoenzymes that can catalyze methanol have not been found, it has long considered hemoenzymes, including cytochrome P450s (P450s), cannot oxidative methane. Herein, we report catalytic by wild-type P450BM3, without any mutagenesis, in presence chemically evolved dummy...
Cytochrome P450s are haem-containing enzymes, catalysing the regio- and stereospecific oxidation of non-activated hydrocarbons. Among these, bacterial P450BM3 is a promising biocatalyst due to its high enzymatic activity. Given significant conformational flexibility this enzyme, understanding protein-substrate interactions associated structural dynamics crucial for designing P450BM3-based biocatalysts. Herein, employing an X-ray free electron laser in combination with freeze-trap...
Significance The short-lived intermediate formed during the reduction of nitric oxide (NO) to nitrous (N 2 O) in denitrification, microbial anaerobic respiration, is a key state for understanding generation mechanism N O, known not only as greenhouse gas but also an ozone-depleting substance on global level. This paper combined state-of-the-art, time-resolved techniques, such flow-flash infrared spectroscopy and X-ray free electron laser-based crystallography, captured P450-type NO reductase...
The addition of perfluorinated fatty acids to the rate accelerating KT2 mutant P450Bm3 resulted in highly active oxidation cyclohexane and benzenes whilst maintaining product selectivity.
Abstract The heme acquisition system A protein secreted by Pseudomonas aeruginosa (HasA p ) can capture several synthetic metal complexes other than heme. crystal structures of HasA harboring revealed only small perturbation the overall structure. An inhibitory effect upon bearing was examined monitoring growth PAO1. bound to iron–phthalocyanine inhibits in presence heme‐bound as an iron source.
The benzylic hydroxylation of non-native substrates was catalysed by cytochrome P450BM3, wherein “decoy molecules” controlled the stereoselectivity reactions.