Fucoxanthin is a carotenoid produced by brown algae and diatoms. This compound has several biological properties such as antioxidant, anti-obesity, anti-diabetic, anticancer, and antimicrobial activities. Unfortunately, until now the latter effect has been poorly confirmed. The aim of this study was an evaluation of fucoxanthin activity against 20 bacterial species. Antimicrobial effect of fucoxanthin was determined by using the agar disc-diffusion and micro-dilution methods. The studied carotenoid acted against 13 bacteria growing in aerobic conditions. It was observed to have a significantly stronger impact on Gram-positive than Gram-negative bacteria. Mean zones of growth inhibition (ZOIs) for Gram-positive bacteria ranged between 9.0 and 12.2 mm, while for Gram-negative were from 7.2 to 10.2 mm. According to the agar disc-diffusion method, the highest activity of fucoxanthin was exhibited against Streptococcus agalactiae (mean ZOI 12.2 mm), Staphylococcus epidermidis (mean ZOI 11.2 mm), and Staphylococcus aureus (mean ZOI 11.0 mm), and in the microdilution test towards Streptococcus agalactiae with the minimal inhibitory concentration (MIC) of 62.5 µg/mL. On the other hand, fucoxanthin was not active against strict anaerobic bacteria.