The Rieske Oxygenase SnoT Catalyzes 2′′‐Hydroxylation of l ‐Rhodosamine in Nogalamycin Biosynthesis

ChemBioChem ◽  
2020 ◽  
Vol 21 (21) ◽  
pp. 3062-3066 ◽  
Author(s):  
Benjamin Nji Wandi ◽  
Vilja Siitonen ◽  
Kaisa Palmu ◽  
Mikko Metsä‐Ketelä
Keyword(s):  
Rieske Oxygenase

scholarly journals Further Studies on the 3-Ketosteroid 9α-Hydroxylase of Rhodococcus ruber Chol-4, a Rieske Oxygenase of the Steroid Degradation Pathway

2021 ◽  
Vol 9 (6) ◽  
pp. 1171
Author(s):  
Sara Baldanta ◽  
Juana María Navarro Llorens ◽  
Govinda Guevara
Keyword(s):  
Cholesterol Metabolism ◽  
Genetic Regulation ◽  
Degradation Pathway ◽  
Rhodococcus Ruber ◽  
Promoter Regions ◽  
Steroid Nucleus ◽  
Steroid Substrate ◽  
Fine Regulation ◽  
Rieske Oxygenase ◽  
Steroid Catabolism

The biochemistry and genetics of the bacterial steroid catabolism have been extensively studied during the last years and their findings have been essential to the development of biotechnological applications. For instance, metabolic engineering of the steroid-eater strains has allowed to obtain intermediaries of industrial value. However, there are still some drawbacks that must be overcome, such as the redundancy of the steroid catabolism genes in the genome and a better knowledge of its genetic regulation. KshABs and KstDs are key enzymes involved in the aerobic breakage of the steroid nucleus. Rhodococcus ruber Chol-4 contains three kshAs genes, a single kshB gene and three kstDs genes within its genome. In the present work, the growth of R. ruber ΔkshA strains was evaluated on different steroids substrates; the promoter regions of these genes were analyzed; and their expression was followed by qRT-PCR in both wild type and ksh mutants. Additionally, the transcription level of the kstDs genes was studied in the ksh mutants. The results show that KshA2B and KshA1B are involved in AD metabolism, while KshA3B and KshA1B contribute to the cholesterol metabolism in R. ruber. In the kshA single mutants, expression of the remaining kshA and kstD genes is re-organized to survive on the steroid substrate. These data give insight into the fine regulation of steroid genes when several isoforms are present.

Dicamba Monooxygenase: Structural Insights into a Dynamic Rieske Oxygenase that Catalyzes an Exocyclic Monooxygenation

2009 ◽  
Vol 392 (2) ◽  
pp. 481-497 ◽  
Author(s):  
Robert L. D'Ordine ◽  
Timothy J. Rydel ◽  
Michael J. Storek ◽  
Eric J. Sturman ◽  
Farhad Moshiri ◽  
...  
Keyword(s):  
Rieske Oxygenase ◽  
Structural Insights

scholarly journals Stereospecific Synthesis of 23-Hydroxyundecylprodiginines and Analogues and Conversion to Antimalarial Premarineosins via a Rieske Oxygenase Catalyzed Bicyclization

2014 ◽  
Vol 79 (23) ◽  
pp. 11674-11689 ◽  
Author(s):  
Papireddy Kancharla ◽  
Wanli Lu ◽  
Shaimaa M. Salem ◽  
Jane Xu Kelly ◽  
Kevin A. Reynolds
Keyword(s):  
Stereospecific Synthesis ◽  
Rieske Oxygenase

scholarly journals The Sterol-C7 Desaturase from the Ciliate Tetrahymena thermophila Is a Rieske Oxygenase, Which Is Highly Conserved in Animals

2013 ◽  
Vol 30 (7) ◽  
pp. 1630-1643 ◽  
Author(s):  
Sebastián R. Najle ◽  
Alejandro D. Nusblat ◽  
Clara B. Nudel ◽  
Antonio D. Uttaro
Keyword(s):  
Tetrahymena Thermophila ◽  
Rieske Oxygenase

scholarly journals Regio- and stereodivergent antibiotic oxidative carbocyclizations catalysed by Rieske oxygenase-like enzymes

2011 ◽  
Vol 3 (5) ◽  
pp. 388-392 ◽  
Author(s):  
Paulina K. Sydor ◽  
Sarah M. Barry ◽  
Olanipekun M. Odulate ◽  
Francisco Barona-Gomez ◽  
Stuart W. Haynes ◽  
...  
Keyword(s):  
Rieske Oxygenase

scholarly journals Design principles for site-selective hydroxylation by a Rieske oxygenase

2022 ◽  
Vol 13 (1) ◽  
Author(s):  
Jianxin Liu ◽  
Jiayi Tian ◽  
Christopher Perry ◽  
April L. Lukowski ◽  
Tzanko I. Doukov ◽  
...  
Keyword(s):  
Structural Information ◽  
Diverse Range ◽  
Bond Functionalization ◽  
Site Selectivity ◽  
Site Selective ◽  
Common Architecture ◽  
Rieske Oxygenase ◽  
Rieske Oxygenases ◽  
Native Site ◽  
Selective Hydroxylation

AbstractRieske oxygenases exploit the reactivity of iron to perform chemically challenging C–H bond functionalization reactions. Thus far, only a handful of Rieske oxygenases have been structurally characterized and remarkably little information exists regarding how these enzymes use a common architecture and set of metallocenters to facilitate a diverse range of reactions. Herein, we detail how two Rieske oxygenases SxtT and GxtA use different protein regions to influence the site-selectivity of their catalyzed monohydroxylation reactions. We present high resolution crystal structures of SxtT and GxtA with the native β-saxitoxinol and saxitoxin substrates bound in addition to a Xenon-pressurized structure of GxtA that reveals the location of a substrate access tunnel to the active site. Ultimately, this structural information allowed for the identification of six residues distributed between three regions of SxtT that together control the selectivity of the C–H hydroxylation event. Substitution of these residues produces a SxtT variant that is fully adapted to exhibit the non-native site-selectivity and substrate scope of GxtA. Importantly, we also found that these selectivity regions are conserved in other structurally characterized Rieske oxygenases, providing a framework for predictively repurposing and manipulating Rieske oxygenases as biocatalysts.

scholarly journals Artificial Light‐Harvesting Complexes Enable Rieske Oxygenase Catalyzed Hydroxylations in Non‐Photosynthetic cells

2020 ◽  
Vol 59 (10) ◽  
pp. 3982-3987 ◽  
Author(s):  
F. Feyza Özgen ◽  
Michael E. Runda ◽  
Bastien O. Burek ◽  
Peter Wied ◽  
Jonathan Z. Bloh ◽  
...  
Keyword(s):  
Light Harvesting ◽  
Artificial Light ◽  
Light Harvesting Complexes ◽  
Rieske Oxygenase ◽  
Photosynthetic Cells

scholarly journals Engineering Burkholderia xenovorans LB400 BphA through Site-Directed Mutagenesis at Position 283

2020 ◽  
Vol 86 (19) ◽  
Author(s):  
Junde Li ◽  
Jun Min ◽  
Yuan Wang ◽  
Weiwei Chen ◽  
Yachao Kong ◽  
...  
Keyword(s):  
Steady State ◽  
Substrate Specificity ◽  
Structural Features ◽  
Site Directed Mutagenesis ◽  
Burkholderia Xenovorans ◽  
Burkholderia Xenovorans Lb400 ◽  
Steady State Kinetics ◽  
Steady State Kinetic ◽  
Degradation Activity ◽  
Rieske Oxygenase

ABSTRACT Biphenyl dioxygenase (BPDO), which is a Rieske-type oxygenase (RO), catalyzes the initial dioxygenation of biphenyl and some polychlorinated biphenyls (PCBs). In order to enhance the degradation ability of BPDO in terms of a broader substrate range, the BphAES283M, BphAEp4-S283M, and BphAERR41-S283M variants were created from the parent enzymes BphAELB400, BphAEp4, and BphAERR41, respectively, by a substitution at one residue, Ser283Met. The results of steady-state kinetic parameters show that for biphenyl, the kcat/Km values of BphAES283M, BphAEp4-S283M, and BphAERR41-S283M were significantly increased compared to those of their parent enzymes. Meanwhile, we determined the steady-state kinetics of BphAEs toward highly chlorinated biphenyls. The results suggested that the Ser283Met substitution enhanced the catalytic activity of BphAEs toward 2,3′,4,4′-tetrachlorobiphenyl (2,3′,4,4′-CB), 2,2′,6,6′-tetrachlorobiphenyl (2,2′,6,6′-CB), and 2,3′,4,4′,5-pentachlorobiphenyl (2,3′,4,4′,5-CB). We compared the catalytic reactions of BphAELB400 and its variants toward 2,2′-dichlorobiphenyl (2,2′-CB), 2,5-dichlorobiphenyl (2,5-CB), and 2,6-dichlorobiphenyl (2,6-CB). The biochemical data indicate that the Ser283Met substitution alters the orientation of the substrate inside the catalytic site and, thereby, its site of hydroxylation, and this was confirmed by docking experiments. We also assessed the substrate ranges of BphAELB400 and its variants with degradation activity. BphAES283M and BphAEp4-S283M were clearly improved in oxidizing some of the 3-6-chlorinated biphenyls, which are generally very poorly oxidized by most dioxygenases. Collectively, the present work showed a significant effect of mutation Ser283Met on substrate specificity/regiospecificity in BPDO. These will certainly be meaningful elements for understanding the effect of the residue corresponding to position 283 in other Rieske oxygenase enzymes. IMPORTANCE The segment from positions 280 to 283 in BphAEs is located at the entrance of the catalytic pocket, and it shows variation in conformation. In previous works, results have suggested but never proved that residue Ser283 of BphAELB400 might play a role in substrate specificity. In the present paper, we found that the Ser283Met substitution significantly increased the specificity of the reaction of BphAE toward biphenyl, 2,3′,4,4′-CB, 2,2′,6,6′-CB, and 2,3′,4,4′,5-CB. Meanwhile, the Ser283Met substitution altered the regiospecificity of BphAE toward 2,2′-dichlorobiphenyl and 2,6-dichlorobiphenyl. Additionally, this substitution extended the range of PCBs metabolized by the mutated BphAE. BphAES283M and BphAEp4-S283M were clearly improved in oxidizing some of the more highly chlorinated biphenyls (3 to 6 chlorines), which are generally very poorly oxidized by most dioxygenases. We used modeled and docked enzymes to identify some of the structural features that explain the new properties of the mutant enzymes. Altogether, the results of this study provide better insights into the mechanisms by which BPDO evolves to change and/or expand its substrate range and its regiospecificity.

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