Metabolic profiling of dormant Mycolicibacterium smegmatis cells’ reactivation reveals a gradual assembly of metabolic processes

Shotgun proteomic profiling of dormant, ‘non-culturable’Mycobacterium tuberculosis

“…A) Pathway visualization, based on KO annotation: the detected enzymes (either stored in the dormant state, or those whose abundancy decreased) were placed on a metabolic map and the corresponding processes highlighted – orange for enzymes stored in dormancy (FC > 0), dark blue for degraded enzymes (FC < 0); line thickness corresponds to statistical significance – the thickest lines depict statistically valuable processes; B) GO enrichment analysis for the pathways assembled by the enzymes stored in dormancy; C) reconstruction of central carbon metabolism pathways of dormant M. tuberculosis cells – Z-scores are mapped on the corresponding Rv coding enzymes; D) mycothiol salvage pathway; E) reconstruction of ‘ de novo ’ UMP pathway. UMP accumulation in the dormant state was similarly recently observed in the same model conditions in M. smegmatis cells (14). …”

“…The classical flow of the glycolytic pathway is hampered by deficient glyceraldehyde-3-phosphate dehydrogenase (Rv1436, p < 0.05, log 2 FC = −0.43) and phosphoglycerate kinase (Rv1437, p < 0.05, log 2 FC = −0.61); however, there is a bypass, consisting of the formation of glycerone phosphate, which can further spontaneously convert into methylglyoxal, which in further detoxification pathways leads to pyruvate through lactate formation. Similarly, all enzymes of glycerol metabolism were detected – the catabolism of glycerol or glycerol-3-phosphate is conjugated with the toxic carbonyl compounds methylglyoxal and lactaldehyde (the latter was detected experimentally for M. smegmatis (14)). However, functioning aldehyde dehydrogenases (Rv0147, Rv2858c) or glyoxylase (Rv0577) detoxify these intermediates, ultimately to lactate.…”

“…The latter can be converted to fumarate by the detected succinate dehydrogenase (Rv3318, Rv0247c). If this pathway is functioning, the accumulation of organic acids (contributing to sustainment of the energized level of transmembrane potential in dormancy (46, 47)) in the cells and medium should be expected; that, however, was found experimentally for dormant cells of M. smegmatis , obtained in the same model of self-acidification of growth medium (14). Contributing to the accumulation of these acids and to detoxification of surplus acetyl-CoA is the glyoxylate pathway, whose enzymes isocitrate lyase (Rv0467) and malate synthase (Rv1837c) were found to be accumulated in dormant cells.…”

Corynebacterium jeikeium Dormant Cell Formation and Photodynamic Inactivation

“…In this study, we ascribe conditions needed for corynebacterial cells to transition to a state of dormancy, which was accompanied by: (1) the formation of stress-resistant forms intended for longterm survival that have been identified in many spore-forming and non-spore-forming bacteria (Zhang, 2004;Mulyukin et al, 2010Mulyukin et al, , 2014Lennon and Jones, 2011) and (2) the acquisition of an NC state during the prolonged incubation of post-stationary cultures, which was previously described for Micrococcus luteus , Rhodococcus rhodochrous (Shleeva et al, 2002) and mycobacteria (M. tuberculosis and M. smegmatis) (Shleeva et al, 2003(Shleeva et al, , 2004(Shleeva et al, , 2011(Shleeva et al, , 2015Kudykina et al, 2011). The dormant forms of mycobacteria, which form when the external medium undergoes gradual acidification, can be distinguished from active bacteria based on their distinct proteomic (Trutneva et al, 2018(Trutneva et al, , 2020 and metabolomic (Nikitushkin et al, 2020) profiles.…”

PE_PGRS31-S100A9 Interaction Promotes Mycobacterial Survival in Macrophages Through the Regulation of NF-κB-TNF-α Signaling and Arachidonic Acid Metabolism

“…Ms is a rapidly growing bacterium (Gupta et al, 2018;Yamada et al, 2018;Nikitushkin et al, 2020) that is frequently used as a substitute for M. tb in studies (Doddam et al, 2019;Kaur and Kaur, 2019;Maslov et al, 2019;Wang et al, 2019). The full-length Rv1768 gene (1768 bp) was cloned into the mycobacterial shuttle vector pMV261 and transformed into Ms to construct a recombinant Ms expressing Rv1768 [Ms (pMV-1768)] (Supplementary Figures S1F,G).…”