Assessment of Metabolic Changes in Mycobacterium smegmatis Wild-Type and alr Mutant Strains: Evidence of a New Pathway of d-Alanine Biosynthesis
Open accessAbstract
In mycobacteria, d -alanine is an essential precursor for peptidoglycan biosynthesis. The only confirmed enzymatic pathway to form d -alanine is through the racemization of l -alanine by alanine racemase (Alr, EC 5.1.1.1). Nevertheless, the essentiality of Alr in Mycobacterium tuberculosis and Mycobacterium smegmatis for cell survivability in the absence of d -alanine has been a point of controversy with contradictory results reported in the literature. To address this issue, we examined the effects of alr inactivation on the cellular metabolism of M. smegmatis . The M. smegmatis alr insertion mutant TAM23 exhibited essentially identical growth to wild-type mc 2 155 in the absence of d -alanine. NMR metabolomics revealed drastically distinct phenotypes between mc 2 155 and TAM23. A metabolic switch was observed for TAM23 as a function of supplemented d -alanine. In the absence of d -alanine, the metabolic response directed carbon through an unidentified transaminase to provide the essential d -alanine required for survival. The process is reversed when d -alanine is available, in which the d -alanine is directed to peptidoglycan biosynthesis. Our results provide further support for the hypothesis that Alr is not an essential function of M. smegmatis and that specific Alr inhibitors will have no bactericidal action.
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NMR was employed to investigate the effect of alr inactivation on Mycobacterium smegmatis cellular metabolism, solving a long-debated controversy regarding the role of this enzyme in d-alanine biosynthesis.
“For instance, NMR was employed to investigate the effect of alr (alanine racemase) inactivation on Mycobacterium smegmatis cellular metabolism, solving a long-debated controversy regarding the role of this enzyme in d-alanine biosynthesis.147”
From Connecting metabolome and phenotype: recent advances in functional metabolomics tools for the identification of bioactive natural products (2024) · cites this paper as [147]
The Introduction (page not provided) says the authors used NMR metabolomics to investigate the M. smegmatis alr mutant and reports evidence suggesting an alternative route to D-alanine. The Abstract (page not provided) identifies Alr essentiality as a controversy, but says the results provide 'further support' for Alr being nonessential in M. smegmatis; describing the controversy as solved overstates that conclusion. (LLM, conf=0.84)
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Vitale, G. A., Geibel, C., Minda, V., Wang, M., Aron, A. T., & Petras, D. (2024) · Natural Product Reports · doi:10.1039/d3np00050h · Cited by 66 · see citing works
- Authors
- Marshall, D. D., Halouska, S., Zinniel, D. K., Fenton, R. J., Kenealy, K., Chahal, H. K., Rathnaiah, G., Barletta, R. G., & Powers, R.
- Affiliations
- University of Nebraska–Lincoln (US)
- Corresponding author
- Barletta, Raúl G.
- Powers, Robert
- Venue
- Journal of Proteome Research
- Publisher
- American Chemical Society (ACS)
- Published
- 2017-02-14
- Volume
- 16
- Issue
- 3
- Pages
- 1270-1279
- ISSN
- 1535-3893 (ISSN-L)
- 1535-3907
- PMC
- PMC6599645
- OpenAlex
- W2581610548
- Cited by
- 13
- References
- 54
- Citations by year
- 2026: 1 · 2025: 1 · 2024: 1 · 2023: 3 · 2021: 1 · 2020: 1 · 2019: 2 · 2017: 3
- FWCI
- 1.26
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Journal of Proteome Research (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Lincoln (University of Nebraska) (submittedVersion)
- Funders
- National Center for Research Resources (P20 RR-17675)
- National Institute of Food and Agriculture (NEB 39-162)
- American Heart Association (0860033Z)
- National Institute of Allergy and Infectious Diseases (AI087668)
- National Institute of General Medical Sciences (P30 GM103335)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Health Sciences › Medicine › Pharmacology › Microbial Natural Products and Biosynthesis
- Keywords
- Mycobacterium smegmatis, alanine racemase, peptidoglycan biosynthesis, NMR metabolomics, metabolic switch, transaminases, bacterial metabolism, Mycobacterium tuberculosis
- MeSH terms
- Alanine, Alanine Racemase, Transaminases, Bacterial Proteins, Mutation, Peptidoglycan, Mycobacterium smegmatis, Metabolic Networks and Pathways
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