Previous studies showed no improvement in bacterial biomass for Candidatus Puniceispirillum marinum IMCC1322 under light regimes. Nevertheless, in nutrient-replete cultures with higher inoculating cell densities, strain IMCC1322 exhibited proteorhodopsin photoheterotrophy. Increasing both inoculum size and the amino acid pool can eliminate quorum sensing and starvation responses in strain IMCC1322. Light regimes affected IMCC1322 cultures in stationary/death phases, where cellular ATP levels ranged from 0.0331 to 1.74 mM, with ATP/cell ranging from 13.9 to 367 zeptomoles. In nutrient-depleted conditions, strain IMCC1322 may suffer from excessive protons generated by proteorhodopsin under light conditions. IMCC1322 may tolerate excessive periplasmic protons through ATP-dependent proton pumping and protonation of augmented amino acids. Meanwhile, acid stress could also be mitigated by refining membrane permeability through unsaturation and cyclopropanation of phospholipids. Oceanic bacteria such as IMCC1322 and SAR11 preferred anaplerotic TCA cycles over glycolysis and rely on the Entner-Doudoroff (ED) pathway for growth. Although ATP generation is less efficient in the ED pathway, it offers advantages during rapid growth owing to its strong thermodynamic driving force. The metabolism of IMCC1322 favors gluconeogenesis over glycolysis, aligning with the metabolism of SAR11 reported in previous studies. However, the additional light-driven, PR-dependent ATP synthesis in IMCC1322 is expected to be insufficient to support protein turnover after the log phase, as well as in nutrient-limited conditions. Stable isotope measurements showed no significant differences in the inorganic carbon assimilation between constant light and constant dark cultures in late log phase.
Keywords: ATP; Candidatus Puniceispirillum marinum; inoculum density; light regime; nutrient repletion; proteorhodopsin.