Marine seaweeds synthesize a plethora of bioactive metabolites, of which phlorotannins of brown algae currently attract special attention due to their high antibiotic and cytotoxic capacities. Here we measured the minimum inhibitory concentrations (MICs) of several semi-purified phlorotannin preparations of different origins and molecular composition using a set of model unicellular organisms, such as Escherichia coli, Saccharomyces cerevisiae, Chlamydomonas reinhardtii, etc. For the first time, MIC values were evaluated for phlorotannin-enriched extracts of brown algae of the orders Ectocarpales and Desmarestiales. Phlorotannin extracts of Desmarestia aculeata, Fucus vesiculosus, and Ectocarpus siliculosus showed the lowest MIC values against most of the treated organisms (4-25 μg/mL for bacteria and yeast). Analysis of the survival curves of E. coli showed that massive loss of cells started after 3-4 h of exposure. Microalgae were less susceptible to activity of phlorotannin extracts, with the highest MIC values (≥200 µg/mL) measured for Chlorella vulgaris cells. D. aculeata, E. siliculosus, and three fucalean algae accumulate considerable amounts (4-16% of dry weight) of phlorotannins with MIC values similar to those widely used antibiotics. As these species grow abundantly in polar and temperate seas and have considerable biomass, they may be regarded as promising sources of phlorotannins.