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Mycobacterium smegmatis possesses (N,N-dimethyl-4-nitrosoaniline)-dependent (NDMA) methanol dehydrogenase (Mno) to establish methylotrophy by utilizing methanol as the source of both carbon and energy. In this study, we show that Mno forms decamer and has NADPH as the bound cofactor. Interestingly, Mno uses NDMA and not NADP as an electron acceptor in in vitro reactions. We further show that the operon mftAD required for the biosynthesis of mycofactocin, a ribosomally-synthesized electron carrier, is indispensable for the growth of M. smegmatis on methanol. Our data obtained from 2,6-Dichlorophenolindophenol reduction assays also suggest that Mno uses mycofactocin as an in vivo electron acceptor for the oxidation of methanol to formaldehyde. We thus provide here biochemical evidence for mycofactocin as an electron carrier in mycobacterial physiology.
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http://dx.doi.org/10.1016/j.bbrc.2019.07.008 | DOI Listing |
Biochemistry
September 2025
Department of Molecular Nutrition, CSIR-Central Food Technological Research Institute (CFTRI), Mysuru, Karnataka 570020, India.
Chromosome organization and segregation are fundamental processes across all domains of life. In bacteria, the mechanisms governing nucleoid organization remain poorly understood. This study investigates the function of an alternative structural maintenance of chromosomes (SMC) complex, MksBEF, in .
View Article and Find Full Text PDFMicrobiol Spectr
September 2025
Institute of Respiratory Health, Frontiers Science Center for Disease-related Molecular Network, West China Hospital, Sichuan University, Chengdu, China.
Efficient DNA delivery is essential for genetic manipulation of mycobacteria and for dissecting their physiology, pathogenesis, and drug resistance. Although electroporation enables transformation efficiencies exceeding 10⁵ CFU per µg DNA in and , it remains highly inefficient in many nontuberculous mycobacteria (NTM), including . Here, we discovered that NTM such as exhibit exceptional tolerance to ultra-high electric field strengths and that hypertonic preconditioning partially protects cells from electroporation-induced damage.
View Article and Find Full Text PDFInt J Biol Macromol
September 2025
Department of Biomedical Science, Acharya Narendra Dev College, University of Delhi, Govindpuri, Kalkaji, 110019, New Delhi, India. Electronic address:
Mycobacteriophage-encoded LysinB enzymes target mycolyl ester linkages in mycolyl-arabinogalactan-peptidoglycan of mycobacterium hosts and generally exhibit a globular architecture. Here, we present the structural and functional characterization of a novel Mycobacterium fortuitum prophage-encoded modular LysinB (LysinB_MF), which contains the α/β hydrolase domain and a distinct peptidoglycan-binding domain (PGBD). The enzyme's active site features the conserved Ser-Asp-His catalytic triad common to esterases and forms a funnel-like topology.
View Article and Find Full Text PDFJ Med Chem
September 2025
Xiangya International Academy of Translational Medicine, Central South University, Changsha, Hunan 410013, China.
KasA and KasB are promising drug targets against and infectious nontuberculous mycobacteria, while most lead compounds are in the preclinical development stage. Herein, a platensimycin (PTM) analogue library consisting of 340 members was first screened to identify 46 PTM thioethers with superior activity compared to that of PTM against . Next, 19 PTM thioethers were chosen and semisynthesized from PTM oxirane (), together with seven PTM ether derivatives and 6-ido, 6-bromo-, and 6-thiocyanato PTM.
View Article and Find Full Text PDFInt J Biol Macromol
September 2025
Biochemistry and Structural Biology Division, CSIR-Central Drug Research Institute, Lucknow, India; Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002, India. Electronic address:
The nucleotide excision repair (NER) pathway in Mycobacterium tuberculosis (Mtb) is important for DNA damage repair and bacterial survival under stress, yet specific inhibitors targeting its components remain scarce. Here, we targeted the UvrB protein, a central component of the Mtb UvrABC NER pathway, and identified novel small molecule inhibitors against its nucleotide binding domain (NBD). Using in silico structure-based screening involving the Maybridge library (~54,000 compounds), Molecular dynamics (MD) simulations, and Biolayer interferometry (BLI), we identified four potent inhibitors: SPB08143, RJC04069, NRB00936, and DP00786 with IC50 values of 9.
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