Biological N₂ fixation was key to the expansion of life on early Earth. The N₂-fixing microorganisms and the nitrogenase type used in the Proterozoic are unknown, although it has been proposed that the canonical molybdenum-nitrogenase was not used due to low molybdenum availability. We investigate N₂ fixation in Lake Cadagno, an analogue system to the sulfidic Proterozoic continental margins, using a combination of biogeochemical, molecular and single cell techniques. In Lake Cadagno, purple sulfur bacteria (PSB) are responsible for high N₂ fixation rates, to our knowledge providing the first direct evidence for PSB in situ N₂ fixation. Surprisingly, no alternative nitrogenases are detectable, and N₂ fixation is exclusively catalyzed by molybdenum-nitrogenase. Our results show that molybdenum-nitrogenase is functional at low molybdenum conditions in situ and that in contrast to previous beliefs, PSB may have driven N₂ fixation in the Proterozoic ocean.

Lakes cover a global area that is about 35 times smaller than the oceans, but carbon burial in lakes and oceans are on the same order of magnitude. Hence, understanding the processes for such high organic carbon burial in lacustrine systems is essential. We applied proxies typically used for marine environments including amino acid (AA) content and their nitrogen stable isotope composition to the water columns and sediments of three lakes that differ in their trophic states and depositions rates of sedimentary terrestrial organic matter. Additionally, we used carbon isotope fingerprinting of AAs to characterise their sources and fates. We show that this set of proxies tracks sources and degradation processes in eutrophic lakes with high sedimentary total organic carbon and nitrogen content. Those lakes also have a high total hydrolysable amino acid (THAA) content as well as higher pigment concentrations. While the Chlorin degradation Index (CI) showed increasing values with depth, the patterns were less consistent for the Degradation Index (DI). In general, all parameters failed to describe degradation in the most oligotrophic lake due to the very low organic carbon and nitrogen content in the sediment. We show that AAs are mostly of autochthonous origin and that AA contribute 5-45 % to the organic material in plankton, POM, and sediment. Proxies based on AA for bacterial reworking (ΣV) or trophic level (TL) showed increasing values in the water column but relatively stable values in the sediments. Furthermore, we show that methanotrophic bacteria led to increased values for the bacterial reworking proxy (ΣV) and extraordinarily low δ¹³C AA values (-30 to -60 ‰).

In marine and freshwater oxygen-deficient zones, the remineralization of sinking organic matter from the photic zone is central to driving nitrogen loss. Deep blooms of photosynthetic bacteria, which form the suboxic/anoxic chlorophyll maximum (ACM), widespread in aquatic ecosystems, may also contribute to the local input of organic matter. Yet, the influence of the ACM on nitrogen and carbon cycling remains poorly understood. Using a suite of stable isotope tracer experiments, we examined the transformation of nitrogen and carbon under an ACM (comprising of Chlorobiaceae and Synechococcales) and a non-ACM scenario in the anoxic zone of Lake Tanganyika. We find that the ACM hosts a tight coupling of photo/litho-autotrophic and heterotrophic processes. In particular, the ACM was a hotspot of organic matter remineralization that controlled an important supply of ammonium driving a nitrification-anammox coupling, and thereby played a key role in regulating nitrogen loss in the oxygen-deficient zone.