Vitamin K₂ mediates electron transport from NADH dehydrogenase 2 to bd-type quinol oxidase in Lacticaseibacillus rhamnosus CM MSU 529

Тo activate respiratory metabolism Lacticaseibacillus rhamnosus CM MSU 529 was grown in a batch culture under intensive aeration in the presence of 38 μM hemin and 18 μM vitamin K₂ as a source of menaquinone. Unsupplemented aerobic culture served as a control. Supplementation of the growth medium with hemin or menaquinone separately had no significant effect on culture growth. Biomass concentration of 2,86 ± 0,05 g dw cells/l and the yield coefficient for biomass Y_P/S of 25,6 ± 1,5 g dw cells/mol glucose consumed were determined after 24 h and 18 h of cultivation under respiratory conditions (hemin + K₂) respectively. Both values were 27% higher compared to those for aerobic conditions. Spectral analysis revealed the presence of cytochromes b- and d-type in membranes of L. rhamnosus CM MSU 529. The activity of bacterial electron transport chain was investigated by polarographic technique. Membrane preparations obtained from cells grown aerobically on hemin-containing medium intensively consumed oxygen in the presence of 1 mM NADH. Addition of 0.2 mM menaquinone to reaction mixture caused the increase of NADH oxidation rate by 4.6 fold. Enzymes presumably involved in NADH oxidation by membranes were identified using MALDI-TOF MS/MS: pyridine nucleotide-disulfide oxidoreductase (Nox-2), NADH dehydrogenase 2 (Ndh-2), and ubiquinol oxidase bd subunit I (CydA). Thus, during NADH oxidation 80% of electron transport from NADH to oxygen went via Ndh-2, menaquinone, bd-type quinol oxidase and only 20% – via Nox-2. The study presents experimental evidence for electron transport chain functioning in L. rhamnosus CM MSU 529 during aerobic cultivation with hemin and menaquinone. The NADH oxidation rates of membrane preparations of lactic acid bacteria were measured for the first time. The property of exogenous menaquinone to transfer electrons from Ndh-2 to bd-type quinol oxidase was demonstrated for the first time in vitro in lactic acid bacteria.

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