The Redox Cofactor F 420 Protects Mycobacteria from Diverse Antimicrobial Compounds and Mediates a Reductive Detoxification System
Open accessAbstract
A defining feature of mycobacterial redox metabolism is the use of an unusual deazaflavin cofactor, F 420 . This cofactor enhances the persistence of environmental and pathogenic mycobacteria, including after antimicrobial treatment, although the molecular basis for this remains to be understood. In this work, we explored our hypothesis that F 420 enhances persistence by serving as a cofactor in antimicrobial-detoxifying enzymes. To test this, we performed a series of phenotypic, biochemical, and analytical chemistry studies in relation to the model soil bacterium Mycobacterium smegmatis . Mutant strains unable to synthesize or reduce F 420 were found to be more susceptible to a wide range of antibiotic and xenobiotic compounds. Compounds from three classes of antimicrobial compounds traditionally resisted by mycobacteria inhibited the growth of F 420 mutant strains at subnanomolar concentrations, namely, furanocoumarins (e.g., methoxsalen), arylmethanes (e.g., malachite green), and quinone analogues (e.g., menadione). We demonstrated that promiscuous F 420 H 2 -dependent reductases directly reduce these compounds by a mechanism consistent with hydride transfer. Moreover, M. smegmatis strains unable to make F 420 H 2 lost the capacity to reduce and detoxify representatives of the furanocoumarin and arylmethane compound classes in whole-cell assays. In contrast, mutant strains were only slightly more susceptible to clinical antimycobacterials, and this appeared to be due to indirect effects of F 420 loss of function (e.g., redox imbalance) rather than loss of a detoxification system. Together, these data show that F 420 enhances antimicrobial resistance in mycobacteria and suggest that one function of the F 420 H 2 -dependent reductases is to broaden the range of natural products that mycobacteria and possibly other environmental actinobacteria can reductively detoxify. IMPORTANCE This study reveals that a unique microbial cofactor, F 420 , is critical for antimicrobial resistance in the environmental actinobacterium Mycobacterium smegmatis . We show that a superfamily of redox enzymes, the F 420 H 2 -dependent reductases, can reduce diverse antimicrobials in vitro and in vivo . M. smegmatis strains unable to make or reduce F 420 become sensitive to inhibition by these antimicrobial compounds. This suggests that mycobacteria have harnessed the unique properties of F 420 to reduce structurally diverse antimicrobials as part of the antibiotic arms race. The F 420 H 2 -dependent reductases that facilitate this process represent a new class of antimicrobial-detoxifying enzymes with potential applications in bioremediation and biocatalysis.
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Mycobacteria encode multiple flavin/deazaflavin oxidoreductases and hydride transferases to survive these stresses.
“encode multiple flavin/deazaflavin oxidoreductases and hydride transferases to survive these stresses (171–174)”
From Mycobacterium smegmatis: The Vanguard of Mycobacterial Research (2023) · cites this paper as [173]
Mycobacteria can metabolize hydrogen gas.
“Studies using the M. smegmatis toolbox have shown that mycobacteria can metabolize hydrogen gas, detoxify carbon monoxide, and encode multiple flavin/deazaflavin oxidoreductases and hydride transferases to survive these stresses (171–174).”
From Mycobacterium smegmatis: The Vanguard of Mycobacterial Research (2023) · cites this paper as [173]
The Results and Discussion (p. 7) address F420-dependent detoxification, not hydrogen-gas metabolism. A References entry (pp. 8–9) names a separate study of atmospheric H2 scavenging by a soil actinobacterium, but our copy does not state that mycobacteria metabolize hydrogen gas. Searches for hydrogenases and hydrogen oxidation found no passage establishing the claim. (LLM, conf=0.70)
Experimental — accuracy not yet measuredMycobacteria can detoxify carbon monoxide.
“mycobacteria can metabolize hydrogen gas, detoxify carbon monoxide, and encode multiple flavin/deazaflavin oxidoreductases and hydride transferases to survive these stresses (171–174)”
From Mycobacterium smegmatis: The Vanguard of Mycobacterial Research (2023) · cites this paper as [173]
The Results and Discussion (p. 7) describe mycobacterial detoxification of antimicrobial compounds, not carbon monoxide. Searches for carbon monoxide detoxification and CO oxidation found no passage in our copy addressing the attributed claim. (LLM, conf=0.75)
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Mycobacterium smegmatis: The Vanguard of Mycobacterial Research
Sparks, I. L., Derbyshire, K. M., Jacobs, W. R., & Morita, Y. S. (2023) · Journal of Bacteriology · doi:10.1128/jb.00337-22 · Cited by 149 · see citing works
- Authors
- Jirapanjawat, T., Ney, B., Taylor, M. C., Warden, A. C., Afroze, S., Russell, R. J., Lee, B. M., Jackson, C. J., Oakeshott, J. G., Pandey, G., & Greening, C.
- Affiliations
- Australian National University (AU)
- Commonwealth Scientific and Industrial Research Organisation (AU)
- ORCID
- Venue
- Applied and Environmental Microbiology
- Publisher
- American Society for Microbiology
- Published
- 2016-12
- Volume
- 82
- Issue
- 23
- Pages
- 6810-6818
- ISSN
- 0099-2240 (ISSN-L)
- 1098-5336
- PMC
- PMC5103081
- OpenAlex
- W2520057004
- Cited by
- 45
- References
- 47
- Citations by year
- 2026: 4 · 2025: 2 · 2024: 7 · 2023: 5 · 2022: 4 · 2021: 6 · 2020: 7 · 2019: 3 · 2018: 2 · 2017: 4 · 2016: 1
- FWCI
- 3.08
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Applied and Environmental Microbiology (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- ANU Open Research (Australian National University) (submittedVersion)
- Funders
- Australian Research Council (DE120102673, DP130102144)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
- Health Sciences › Medicine › Epidemiology › Mycobacterium research and diagnosis
- Health Sciences › Medicine › Pharmacology › Microbial Natural Products and Biosynthesis
- Keywords
- F420, Mycobacterium smegmatis, antimicrobial resistance, hydride transfer, furocoumarins, 8-methoxypsoralen, malachite green, menadione, actinobacteria, redox metabolism
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