Functional and Structural Characterization of Diverse NfsB Chloramphenicol Reductase Enzymes from Human Pathogens

Mullowney, Michael W.; Maltseva, N.; Endres, M.; Kim, Young Chang; Joachimiak, A.; Crofts, Terence Spencer

doi:10.1128/spectrum.00139-22

Cited by 9 publications

(4 citation statements)

References 52 publications

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“…Strongly depending on the affinity for the substrates, different product distributions are observed for NsfB. Indeed, NfsB is known to contribute to chloramphenicol resistance in Gram-negative bacteria by reduction of chloramphenicol to its corresponding inactive aminochloramphenicol, [54,55] but likewise induces susceptibility of Gram-negative bacteria towards nitrometronidazol by conversion into its hydroxylamine [55] involved in radical processes damaging the bacterial DNA. In our current mechanistic understanding of the overall reaction, the initial reduction of psGSLPEG(NO2)-N3 to the corresponding nitroso compound 22 (only detected in traces, Scheme 4) and subsequently to the hydroxylamine 21 as major reduction product is fast.…”

Section: Resultsmentioning

confidence: 99%

Bioresponsive pseudoGlucosinolates (psGSLs) release Isothiocyanates (ITCs) in the Presence of Nitroreductases

Jimidar,

Ganskow,

Kagho

et al. 2024

Preprint

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Glucosinolates (GSLs) are secondary metabolites produced as part of an herbivore defence system in plants of the order Brassicales. GSLs release isothiocyanates (ITCs) upon activation by the myrosinase. Beyond their herbivore feeding deterrent properties, these ITCs have multiple interesting bioactivities. However, their release is limited by the presence of myrosinase. Here, we report the concept of pseudoglucosinolates (psGSLs) hijacking the natural release mechanism of GSLs for the release of ITCs and adapting it to nitroreductase as triggering enzymes. We provide the proof-of-concept for nitroreductase-responsive psGSLs and demonstrate their potential for peptide labelling and ITC-prodrug approaches.

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Section: Resultsmentioning

confidence: 99%

Bioresponsive pseudoGlucosinolates (psGSLs) release Isothiocyanates (ITCs) in the Presence of Nitroreductases

Jimidar,

Ganskow,

Kagho

et al. 2024

Preprint

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“…Resistance may emerge directly from elements with other metabolic functions or indicate a shared common ancestor. − The recent finding that soil bacteria are capable of subsisting on antibiotics as the sole carbon source further supports a close association between antibiotic resistance and bacterial metabolism. − Given the astounding number of bacterial cells on the planet (>10 30 ), the variety of metabolic enzymes that can serve as resistance elements, and the rapidity of bacterial cell growth, we believe that environmental bacteria capable of metabolizing aminoglycosides may also be widespread. Analyzing these bacteria could reveal aminoglycoside-degrading enzymes.…”

Section: Introductionmentioning

confidence: 84%

Deglycosylation Inactivation Initiated by a Novel Periplasmic Dehydrogenase Complex Provides a Novel Strategy for Eliminating the Recalcitrant Antibiotic Kanamycin

Chen

Liu

Chen

et al. 2023

Environ. Sci. Technol.

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Biodegradation using enzyme-based systems is a promising approach to minimize antibiotic loads in the environment. Aminoglycosides are refractory antibiotics that are generally considered non-biodegradable. Here, we provide evidence that kanamycin, a common aminoglycoside antibiotic, can be degraded by an environmental bacterium through deglycosylation of its 4′-amino sugar. The unprecedented deglycosylation inactivation of kanamycin is initiated by a novel periplasmic dehydrogenase complex, which we designated AquKGD, composed of a flavin adenine dinucleotidedependent dehydrogenase (AquKGDα) and a small subunit (AquKGDγ) containing a twin-arginine signal sequence. We demonstrate that the formation of the AquKGDα−AquKGDγ complex is required for both the degradation activity of AquKGD and its translocation into the periplasm. Native AquKGD was successfully expressed in the periplasmic space of Escherichia coli, and physicochemical analysis indicated that AquKGD is a stable enzyme. AquKGD showed excellent degradation performance, and complete elimination of kanamycin from actual kanamycin manufacturing waste was achieved with immobilized AquKGD. Ecotoxicity and cytotoxicity tests suggest that AquKGD-mediated degradation produces less harmful degradation products. Thus, we propose a novel enzymatic antibiotic inactivation strategy for effective and safe treatment of recalcitrant kanamycin residues.

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“…and Haemophilus influenzae , has been proposed in multiple reports in the early stages (Smith et al, 2007). However, only Mullowney et al (2022) identified the nitroreductase NfsB from the strain H. influenzae , which confers resistance to CHL by reducing the nitro group to the corresponding amine group.…”

Section: Introductionmentioning

confidence: 99%

ChlOR, a GMC family oxidoreductase that evolved independently from the actinomycete, confers resistance to amphenicol antibiotics

Qian,

Cheng,

Lai

et al. 2023

Environmental Microbiology

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Overuse of the amphenicol antibiotics chloramphenicol (CHL) and thiamphenicol (TAP) poses a great threat to ecosystem safety and human health. The strain, Nocardioides sp. LMS‐CY, Nocardioides sp. QY071 and Nocardioides sp. L‐11A, classified as a gram‐positive actinomycete, harbours a complete CHL metabolic pathway. However, the metabolic genes (clusters) involved in the entire pathway in gram‐positive actinomycetes are still limited. Here, chlORLMS, chlORQY071 and chlORL‐11A completely from the actinomycete Nocardioides spp. were found to act on the C1‐OH of the CHL/TAP side chain, directly converting CHL/TAP to 4‐nitrobenzaldehyde (PNBD)/4‐methylsulfonyl benzaldehyde (PMBD) and transforming PNBD/PMBD into 4‐nitrobenzyl alcohol (PNBM)/4‐methylsulfonyl phenyl methanol (PMBM). Furthermore, oxidoreductases can transform PNBM into 4‐nitrobenzoate (PNBA). The oxidoreductases ChlORLMS, ChlORQY071 and ChlORL‐11A were all classified as cellobiose dehydrogenases from the glucose methanol choline (GMC) family. Based on the Swiss‐Prot database, ChlORQY071 exhibited a lower identity (27.12%–35.10% similarity) with the reported oxidoreductases. Enzymatic and molecular docking analyses showed that ChlORQY071 and ChlORL‐11A from the two similar genomes were remarkably more effective in metabolizing CHL than ChlORLMS. Overall, the detailed resistance mechanism of CHL/TAP by actinomycete strains isolated from soil and livestock manure will provide insights into the occurrence of CHL/TAP resistance genes in the environment, resistance risk and bioremediation of CHL/TAP‐contaminated environments.

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Functional and Structural Characterization of Diverse NfsB Chloramphenicol Reductase Enzymes from Human Pathogens

Cited by 9 publications

References 52 publications

Bioresponsive pseudoGlucosinolates (psGSLs) release Isothiocyanates (ITCs) in the Presence of Nitroreductases

Bioresponsive pseudoGlucosinolates (psGSLs) release Isothiocyanates (ITCs) in the Presence of Nitroreductases

Deglycosylation Inactivation Initiated by a Novel Periplasmic Dehydrogenase Complex Provides a Novel Strategy for Eliminating the Recalcitrant Antibiotic Kanamycin

ChlOR, a GMC family oxidoreductase that evolved independently from the actinomycete, confers resistance to amphenicol antibiotics

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