Ondokuz Mayis University, Engineering Faculty, Department of Food Engineering, 55139, Samsun, Turkey.
Kastamonu University, Faculty of Engineering and Architecture, Department of Food Engineering, 37200, Kastamonu, Turkey.
Cryobiology. 2020 Oct;96:122-129. doi: 10.1016/j.cryobiol.2020.07.007. Epub 2020 Jul 23.
In this study, the response surface methodology was used to optimize the cryoprotective agent (skimmed milk powder, lactose and sucrose) formulation for enhancing the viability of Lactobacillus curvatus N19 during freeze-drying and storage stability of cells freeze-dried by using optimum formulation was evaluated. Our results showed that the most significant cryoprotective agent influencing the viability of L. curvatus N19 to freezing and freeze-drying was sucrose and skim milk, respectively. The optimal formulation of cryoprotective agents was 20 g/100 mL skim milk, 3.57 g/100 mL lactose and 10 g/100 mL sucrose. Using the optimum formulation during freeze-drying, the cell survival was found more than 98%. Under the optimal conditions, although only storage of the cells at 4 °C for 6 month retained the maximum stability (8.85 log cfu/g), the employed protectant matrix showed promising results at 25 °C (7.89 log cfu/g). The storage stability of cells under optimized conditions was predicted by accelerated storage test, which was demonstrated that the inactivation rate constant of the freeze-dried L. curvatus N19 powder was 9.74 × 10 1/d for 4 °C and 2.08 × 10 1/d for 25 °C. The loss of specific acidification activity after the storage at 4 and 25 °C was determined.
在这项研究中,使用响应面法优化了冷冻保护剂(脱脂乳粉、乳糖和蔗糖)配方,以提高干酪乳杆菌 N19 在冷冻干燥过程中的存活率,并评估了使用最佳配方冻干细胞的储存稳定性。我们的结果表明,对干酪乳杆菌 N19 冷冻和冻干存活率影响最大的冷冻保护剂分别是蔗糖和脱脂乳。冷冻保护剂的最佳配方为 20 g/100 mL 脱脂乳、3.57 g/100 mL 乳糖和 10 g/100 mL 蔗糖。在冷冻干燥过程中使用最佳配方,细胞存活率超过 98%。在最佳条件下,尽管只有在 4°C 下储存 6 个月才能保持最大稳定性(8.85 log cfu/g),但所采用的保护基质在 25°C 下显示出有希望的结果(7.89 log cfu/g)。通过加速储存试验预测了优化条件下细胞的储存稳定性,结果表明,4°C 下冻干干酪乳杆菌 N19 粉末的失活速率常数为 9.74×10 1/d,25°C 下为 2.08×10 1/d。确定了在 4 和 25°C 下储存后的特定酸化活性损失。