Banded iron formations (BIFs) exhibit alternating silica- and iron-rich laminae, potentially reflecting the dynamics of the paleo-environments in which they were formed, although the exact mechanism remains unclear. Here the formation of a 2.7–2.9 Ga BIF from Dharwar Craton, India, is deciphered by analyzing the inter-band variations of the redox-sensitive isotope biomarkers, 15N/14N and 56Fe/54Fe. Organic matter with δ15N values as high as appears to be trapped in silica. Iron oxides exhibit systematically positive δ56Fe values, ranging between and . Compared to the iron-rich bands, silica-rich bands, which show higher δ56Fe values, exhibit an order of magnitude higher concentrations of 15N-rich organic nitrogen, normalized by the abundances of its host silica. The presence of 15N-rich organic matter may imply the emergence of a modern-like biological nitrogen cycle that requires the formation of oxidized nitrogen compounds. The higher concentration of 15N-rich organic nitrogen for the silica-rich bands possibly suggests that the photosynthetic activity was higher during the formation periods of these bands. The heavier iron isotope compositions of the silica-rich bands cannot be explained alone by iron oxidation through probable pathways. The relative 56Fe-enrichment in silica-rich bands is explained here by the progressive dissolution of iron oxides to the ocean, through iron reduction by 15N-rich organic matter actively produced at the ocean surface. The formation of iron-rich bands possibly corresponds to periods of reduced biological productivity, when precipitated iron was not effectively dissolved to the ocean. The observed shift in the organic concentration between Fe- and Si-rich bands could be the switch that triggered the BIF laminations. This shift could conceivably represent periodic fluctuations in the oxygen generation, which possibly occurred over periods of millennia, at the dawn of the Great Oxidation Event.