Growth limitation of marine algae due to lack of iron occurs in up to 40% of the global ocean. Despite important advances on the impact of organic compounds on iron biogeochemistry, their roles in controlling iron availability to prokaryotic and eukaryotic phytoplankton remain unclear. Whether algal and bacterial exopolymeric substances (EPS) include organic ligands which may help iron-limited phytoplankton growth remains an unknown. If so, then EPS could relieve phytoplankton iron limitation with implications for the biological carbon pump and hence the regulation of atmospheric CO2. Here we compared the biological impact of algal, bacterial and in situ EPS with model compounds, a siderophore and two saccharides on biological parameters including, iron bioavailability, phytoplankton growth, photo-physiology and community structure. Laboratory and field experiments demonstrated that EPS produced by marine microorganisms are efficient in sustaining biological iron uptake as well as algal growth, and can affect natural phytoplankton community structure. Our data suggest that natural phytoplankton growth enhancement in the presence of EPS was not solely due to highly bioavailable iron forms, but also because EPS contains other micronutrients. Stronger ligands were detected following iron-siderophore enrichments (log KFe′L = 12.0) and weaker ligands were measured in the presence of EPS (log KFe′L = 10.4–11.0). The trend of the conditional stability constants of organic ligands did not seem to be affected as a result of biological activity and photo-chemistry during our four day incubations. The shift in the phytoplankton community observed during our field experiments was not uniformly observed between different sites rendering it difficult to extrapolate which functional group(s) would benefit the most from iron bound to EPS.