Abstract The stability of levan produced by Zymomonas mobilis 113 “S” was investigated during storage of culture liquid at different pH and temperatures. Levan was not degraded if stored in a culture liquid for 120 h at 30 or 25 °C, but at 55 or 60 °C the degree of degradation was inversely proportional to pH. Cell biomass and levan sediment components did not cause the degradation of levan. The influence of organic acids (lactic-, acetic-, and gluconic) and HCl as levan hydrolyzing agents at pH 3.7, temperature 30 °C or 55 °C was investigated in a model solution. The hydrolytic effect was similar in the case of organic acids—ensuring more fructose formation, while HCl ensured higher amounts of fructo-oligosaccharides (FOS) and/or low molecular weight levan. Consequently, acid hydrolysis acts as a main levan degrading factor during storage in a culture liquid.