UMR Procédés Alimentaires et Microbiologiques, University Bourgogne Franche-Comté, AgroSup Dijon, PAM UMR A 02.102, 21000, Dijon, France.
Indigo Therapeutics, 5 rue Salneuve, 75017, Paris, France.
BMC Biotechnol. 2021 Nov 12;21(1):66. doi: 10.1186/s12896-021-00726-2.
Stabilization of freeze-dried lactic acid bacteria during long-term storage is challenging for the food industry. Water activity of the lyophilizates is clearly related to the water availability and maintaining a low a during storage allows to increase bacteria viability. The aim of this study was to achieve a low water activity after freeze-drying and subsequently during long-term storage through the design of a lyoprotectant. Indeed, for the same water content as sucrose (commonly used lyoprotectant), water activity is lower for some components such as whey, micellar casein or inulin. We hypothesized that the addition of these components in a lyoprotectant, with a higher bound water content than sucrose would improve lactobacilli strains survival to long-term storage. Therefore, in this study, 5% whey (w/v), 5% micellar casein (w/v) or 5% inulin (w/v) were added to a 5% sucrose solution (w/v) and compared with a lyoprotectant only composed of 5% sucrose (w/v). Protective effect of the four lyoprotectants was assessed measuring Lactiplantibacillus plantarum CNCM I-4459 survival and water activity after freeze-drying and during 9 months storage at 25 °C.
The addition whey and inulin were not effective in increasing Lactiplantibacillus plantarum CNCM I-4459 survival to long-term-storage (4 log reduction at 9 months storage). However, the addition of micellar casein to sucrose increased drastically the protective effect of the lyoprotectant (3.6 log i.e. 0.4 log reduction at 9 months storage). Comparing to a lyoprotectant containing whey or inulin, a lyoprotectant containing micellar casein resulted in a lower water activity after freeze-drying and its maintenance during storage (0.13 ± 0.05).
The addition of micellar casein to a sucrose solution, contrary to the addition of whey and inulin, resulted in a higher bacterial viability to long-term storage. Indeed, for the same water content as the others lyoprotectants, a significant lower water activity was obtained with micellar casein during storage. Probably due to high bound water content of micellar casein, less water could be available for chemical degradation reactions, responsible for bacterial damages during long-term storage. Therefore, the addition of this component to a sucrose solution could be an effective strategy for dried bacteria stabilization during long-term storage.
对于食品工业来说,在长期储存过程中稳定冻干乳酸菌是一项具有挑战性的工作。冷冻干燥产品的水活度显然与水分可用性有关,在储存过程中保持低水活度可以提高细菌的存活率。本研究的目的是通过设计一种保护剂,在冷冻干燥后和长期储存过程中实现低水活度。事实上,对于与蔗糖(常用的保护剂)相同的含水量,一些成分(如乳清、胶束酪蛋白或菊粉)的水活度更低。我们假设,在保护剂中添加这些具有比蔗糖更高结合水含量的成分,将提高乳酸菌菌株对长期储存的存活率。因此,在这项研究中,5%乳清(w/v)、5%胶束酪蛋白(w/v)或 5%菊粉(w/v)被添加到 5%蔗糖溶液(w/v)中,并与仅由 5%蔗糖(w/v)组成的保护剂进行了比较。通过测量冷冻干燥后和在 25°C 下储存 9 个月期间的植物乳杆菌 CNCM I-4459 的存活率和水活度,评估了四种保护剂的保护效果。
乳清和菊粉的添加并没有有效地提高植物乳杆菌 CNCM I-4459 对长期储存的存活率(在 9 个月储存期时减少了 4 个对数级)。然而,向蔗糖中添加胶束酪蛋白极大地增加了保护剂的保护效果(3.6 个对数级,即在 9 个月储存期时减少了 0.4 个对数级)。与含有乳清或菊粉的保护剂相比,含有胶束酪蛋白的保护剂在冷冻干燥后和储存过程中具有更低的水活度(0.13±0.05)。
与添加乳清和菊粉相反,向蔗糖溶液中添加胶束酪蛋白可显著提高细菌在长期储存中的存活率。事实上,对于与其他保护剂相同的含水量,在储存过程中,使用胶束酪蛋白可以获得显著更低的水活度。可能是由于胶束酪蛋白具有较高的结合水含量,在长期储存过程中,用于化学降解反应的水分较少,这是导致细菌受损的原因。因此,在蔗糖溶液中添加这种成分可能是一种在长期储存过程中稳定冻干细菌的有效策略。