The Rieske non-heme iron oxygenases (Rieske oxygenases) comprise a class of metalloenzymes that are involved in the biosynthesis of complex natural products and the biodegradation of aromatic pollutants. Despite this desirable catalytic repertoire, industrial implementation of Rieske oxygenases has been hindered by the multicomponent nature of these enzymes and their requirement for expensive reducing equivalents in the form of a reduced nicotinamide adenine dinucleotide cosubstrate (NAD(P)H). Fortunately, however, some Rieske oxygenases co-occur with accessory proteins, that through a downstream reaction, recycle the needed NAD(P)H for catalysis. As these pathways and accessory proteins are attractive for bioremediation applications and enzyme engineering campaigns, herein, we describe methods for assembling Rieske oxygenase pathways in vitro. Further, using the TsaMBCD pathway as a model system, in this chapter, we provide enzymatic, spectroscopic, and crystallographic methods that can be adapted to explore both Rieske oxygenases and their co-occurring accessory proteins.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/bs.mie.2024.05.015 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
-Dihydroxylation by Synthetic Iron(III)-Peroxo Intermediates and Rieske Dioxygenases: Experimental and Theoretical Approaches Reveal the Key O-O Bond Activation Step.
J Am Chem Soc
November 2024
Department of Chemistry and Nano Science, Ewha Womans University, Seoul 03760, Korea.
Dioxygen (O) activation by iron-containing enzymes and biomimetic compounds generates iron-oxygen intermediates, such as iron-superoxo, -peroxo, -hydroperoxo, and -oxo, that mediate oxidative reactions in biological and abiological systems. Among the iron-oxygen intermediates, iron(III)-peroxo species are less frequently implicated as active intermediates in oxidation reactions. In this study, we present the combined experimental and theoretical investigations on -dihydroxylation reactions mediated by synthetic mononuclear nonheme iron-peroxo intermediates, demonstrating the importance of supporting ligands and metal centers in activating the peroxo ligand toward the O-O bond homolysis for the -dihydroxylation reactions.
View Article and Find Full Text PDFEfficient sulfamethoxazole biotransformation and detoxification by newly isolated strain Hydrogenophaga sp. SNF1 via a ring ortho-hydroxylation pathway.
J Hazard Mater
December 2024
Key Lab of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, 1799 Jimei Road, Xiamen 361021, PR China; University of Chinese Academy of Sciences, 19A Yuquan Road, Beijing 100049, PR China.
Sulfonamides are frequently detected with high concentrations in various environments and was regarded as a serious environmental risk by fostering the dissemination of antibiotic resistance genes. This study for the first time reported a strain SNF1 affiliated with Hydrogenophaga can efficiently degrade sulfamethoxazole (SMX). Strain SNF1 prefers growing under extra carbon sources and neutral condition, and could degrade 500 mg/L SMX completely within 16 h.
View Article and Find Full Text PDFIntramolecular C-H Oxidation in Iron(V)-oxo-carboxylato Species Relevant in the γ-Lactonization of Alkyl Carboxylic Acids.
ACS Catal
September 2024
Institut de Química Computacional i Catàlisi (IQCC), Departament de Química, Universitat de Girona, C/Ma Aurèlia Capmany 69, 17003 Girona, Catalonia, Spain.
High-valent oxoiron species have been invoked as oxidizing agents in a variety of iron-dependent oxygenases. Taking inspiration from nature, selected nonheme iron complexes have been developed as catalysts to elicit C-H oxidation through the mediation of putative oxoiron(V) species, akin to those proposed for Rieske oxygenases. The addition of carboxylic acids in these iron-catalyzed C-H oxidations has proved highly beneficial in terms of product yields and selectivities, suggesting the direct involvement of iron(V)-oxo-carboxylato species.
View Article and Find Full Text PDFExpression, purification, kinetics, and crystallization of non-heme mononuclear iron enzymes: Biphenyl, Phthalate, and Terephthalate dioxygenases.
Methods Enzymol
September 2024
Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand, India. Electronic address:
Non-heme iron oxygenases constitute a versatile enzyme family that is crucial for incorporating molecular oxygen into diverse biomolecules. Despite their importance, only a limited number of these enzymes have been structurally and functionally characterized. Surprisingly, there remains a significant gap in understanding how these enzymes utilize a typical architecture and reaction mechanism to catalyze a wide range of reactions.
View Article and Find Full Text PDFMethods used to determine the structure of the oxygenase component of naphthalene 1,2 dioxygenase.
Methods Enzymol
September 2024
Department of Biological Sciences, Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, United States. Electronic address:
Rieske non-heme iron oxygenases are ubiquitously expressed in prokaryotes. These enzymes catalyze a wide variety of reactions, including cis-dihydroxylation, mono-hydroxylation, sulfoxidation, and demethylation. They contain a Rieske-type [2Fe-2S] cluster and an active site with a mono-nuclear iron bound to a 2-His carboxylate triad.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!