The effect of various polysaccharides commonly used as thickening or gelling agents in foods on the antibacterial activity of nisin Z was investigated. Addition of 1 g.L−1 sodium alginate in tryptic soy broth resulted in a four-fold increase of minimal inhibitory (MIC) and bactericidal (MBC) concentrations of nisin Z against Listeria innocua ATCC 33090 and Kocuria rhizophila ATCC 9341, while the addition of citrus pectin or various exopolysaccharides (Leuconostoc mesenteroides P35 exopolysaccharides, low (9–11 kDa) or medium (60–76 kDa) molecular weight dextran from Ln. mesenteroides and 2 kefirans (extracted from kefir grains provided by Crokfun or Kefiralia)) did not increase MIC and MBC of nisin Z against these bacteria. Zeta potential determination at pH 7 of the nisin Z, bacteria and polysaccharides allowed us to observe that only nisin Z had a slightly positive zeta potential, while the bacteria, sodium alginate and highly methoxylated citrus pectin zeta potentials were lower than −20 mV and zeta potentials of all exopolysaccharides were between 0 and - 10 mV. It was thus proposed that anionic sodium alginate, which is the only polysaccharide having a lower zeta potential than both bacteria, forms complexes stabilized by electrostatic interactions with oppositely charged nisin Z, thereby limiting the quantity of “free” nisin Z interacting with nisin-susceptible bacteria. This hypothesis was substantiated by the observation of nisin Z - sodium alginate aggregation at pH 7 and the estimation of the apparent affinity of sodium alginate for “free” nisin Z and immobilized nisin Z by partition experiments and by surface plasmon resonance analysis, respectively. This study thus provides useful information for food formulation regarding the effect of different polysaccharides on nisin Z antibacterial activity and some understanding of how interactions between nisin Z, polysaccharides, and nisin Z - sensitive bacteria are explanatory.