Abstract:An ice-nucleating bacterium, designated MACK-4, was isolated from ice-stored mackerel (Scomber australasicus) and identified as a Pseudomonas fluorescens. The optimal temperature and pH for its growth in nutrient broth with 2.5% glycerol (NB-G) were 15°C and 6.5, respectively. The maximal ice-nucleating activity (INA) was obtained after 54 h incubation at 15°C. However, the INA was almost completely lost after 48 h incubation at 25°C or higher. The growth and INA decreased with increase of NaCl added in NB-G within 0.0–4.0%. The INA of MACK-4 was very stable at 5–25°C, pH 4.0–9.5, while that of isolated ice-nucleating matter from MACK-4 was stable at 5–25°C, pH 5.5–9.0.