为了提高干酪乳杆菌CCFM711的冻干存活率及冻干粉的耐酸性,测定了不同耐酸性壁材冻干菌体后的耐酸性,以及菌体在不同种类糖与蛋白质保护下的冻干存活率。将壁材与冻干保护剂复配制备冻干菌粉,测定菌体冻干存活率和耐酸性,并与优化乳化法制备的海藻酸钠微胶囊进行比较,对真空冷冻干燥制备干酪乳杆菌耐酸冻干粉的工艺进行优化。以水苏糖作为冻干保护剂分别复配海藻酸钠、酪蛋白酸钠制备冻干菌粉能够同时提高其冻干存活率与耐酸性。以水苏糖作为冻干保护剂,制备海藻酸钠湿微胶囊能够显著提高菌体在酸性环境中的存活率,但冻干后其耐酸性显著降低,仅为(0.37±0.28)%,不及非微胶囊冻干粉[(19.56±1.83)%]。以水苏糖(100 g/L)作为冻干保护剂,酪蛋白酸钠(10 g/L)为耐酸壁材,菌悬液与水苏糖-酪蛋白酸钠混合溶液以体积比2∶5混合时,其冻干存活率为(59.43±6.28)%,pH 2.5下处理2 h,存活率达(34.16±7.18)%,耐酸性显著高于无壁材冻干粉。该研究可为制备干酪乳杆菌耐酸冻干粉提供指导。
In order to improve the freeze-drying survival rate and the acid resistance of lyophilized powder of Lactobacillus casei CCFM711, cells were lyophilized with different acid-resistant materials, and with sugars and proteins as protectants for freeze-drying. The lyophilized cell powder was then tested and compared with the sodium alginate microcapsules prepared by the optimized emulsification method. Finally, the vacuum freeze-drying process was optimized. The results showed that stachyose combined with sodium alginate and sodium caseinate, respectively, could simultaneously improve the survival rate in freeze-drying and the acid resistance. Using stachyose as protectant for freeze-drying in preparation of sodium alginate wet microcapsules, the survival rate of bacteria in acidic environment could be significantly improved, but the acid resistance was significantly reduced, which was only (0.37±0.28) %, much less than un-coated lyophilized powder (19.56±1.83) %. Stachyose (100g/L) was used as the protectant for freeze-drying, and sodium caseinate (10 g/L) as the optimal wall material. Mixing the bacterial suspension and the protectants at the ratio of 2∶5, a freeze-drying survival rate of (59.43±6.28) % was obtained, while the highest survival rate was (34.16±7.18) % after 2 hours' treatment at pH 2.5, about 8 times of the control. This study provides guidance for the preparation of lyophilized powder of L. casei.
[1] DA C A G, BURITI F C A, DE S C H B, et al. Probiotic cheese: health benefits, technological and stability aspects[J]. Trends in Food Science & Technology, 2009, 20(8): 344-354.
[2] SARAO L K, ARORA M. Probiotics, prebiotics, and microencapsulation: A review[J]. Critical Reviews in Food Science and Nutrition, 2017, 57(2): 344-371.
[3] KOSARAJU S L, WEERAKKODY R, AUGUSTIN M A. In-vitro evaluation of hydrocolloid-based encapsulated fish oil[J]. Food Hydrocolloids, 2009, 23(5): 1 413-1 419.
[4] PARK J H, SARAVANAKUMAR G, KIM K, et al. Targeted delivery of low molecular drugs using chitosan and its derivatives[J]. Advanced Drug Delivery Reviews, 2010, 62(1): 28-41.
[5] 杨佳, 侯占群, 贺文浩, 等. 微胶囊壁材的分类及其性质比较[J]. 食品与发酵工业, 2009, 35(5): 122-127.
[6] 宋娇娇, 王俊国, 田文静, 等. 微胶囊制备工艺对益生菌活性的影响[J]. 中国乳品工业, 2016, 44(1): 25-30.
[7] LIU H, CUI S W, CHEN M, et al. Protective approaches and mechanisms of microencapsulation to the survival of probiotic bacteria during processing, storage and gastrointestinal digestion: a review[J]. Crit Rev Food Sci Nutr, 2019,59(17): 2 863-2 878.
[8] DOHERTY S, GEE V, ROSS R, et al. Development and characterisation of whey protein micro-beads as potential matrices for probiotic protection[J]. Food Hydrocolloids, 2011, 25(6): 1 604-1 617.
[9] GONZALEZ-FERRERO C, IRACHE J M, GONZALEZ-NAVARRO C J. Soybean protein-based microparticles for oral delivery of probiotics with improved stability during storage and gut resistance[J]. Food Chem, 2018, 239: 879-888.
[10] 田文静. 内源乳化法制备植物乳杆菌LIP-1微胶囊及提高其抗冷冻性能的研究[D].呼和浩特:内蒙古农业大学, 2016.
[11] S NCHEZ M T, RUIZ M A, LASSERROT A, et al. An improved ionic gelation method to encapsulate Lactobacillus spp. bacteria: Protection, survival and stability study[J]. Food Hydrocolloids, 2017, 69: 67-75.
[12] YEUNG T W, ARROYO-MAYA I J, MCCLEMENTS D J, et al. Microencapsulation of probiotics in hydrogel particles: enhancing Lactococcus lactis subsp. cremoris LM0230 viability using calcium alginate beads[J]. Food Funct, 2016, 7(4): 1 797-1 804.
[13] IACONELLI C, LEMETAIS G, KECHAOU N, et al. Drying process strongly affects probiotics viability and functionalities[J]. J Biotechnol, 2015, 214: 17-26.
[14] 张国芳, 王婷婷, 刘丽波, 等. 内源乳化法制备干酪乳杆菌微胶囊[J]. 中国乳品工业, 2017, 45(3): 15-20.
[15] ASHWAR B A, GANI A, GANI A, et al. Production of RS4 from rice starch and its utilization as an encapsulating agent for targeted delivery of probiotics[J]. Food Chem, 2018, 239: 287-294.
[16] 曾凤泽, 陈霞, 邵玉宇, 等. 刍议益生菌微胶囊化壁材的分类及性质[J]. 中国乳品工业, 2013, 41(6): 27-30.
[17] WANG W, CHEN M, WU J, et al. Hypothermia protection effect of antifreeze peptides from pigskin collagen on freeze-dried Streptococcus thermophiles and its possible action mechanism[J]. LWT - Food Science and Technology, 2015, 63(2): 878-885.
[18] ANA S CARVALHO, JOANA SILVA, PETER H O, et al. Effects of various sugars added to growth and drying media upon thermotolerance and survival throughout storage of freeze-dried Lactobacillus delbrueckii ssp. bulgaricus[J].Biotechnology Pregress, 2004, 20(1):248-254.
[19] HONGPATTARAKERE T, URAIPAN S. Bifidogenic characteristic and protective effect of saba starch on survival of Lactobacillus plantarum CIF17AN2 during vacuum-drying and storage[J]. Carbohydr Polym, 2015, 117: 255-261.
[20] 朱莹丹. 植物乳杆菌LIP-1微胶囊制备工艺及其性能的研究[D]. 呼和浩特:内蒙古农业大学, 2015.
[21] 程玉霞. 基于外源乳化法的植物乳杆菌微胶囊制备及性能研究[D]. 杭州:浙江工商大学, 2012.
[22] 朱永明, 杜玲玲, 李晓东, 等. 胶囊化鼠李糖乳杆菌在模拟胃肠道中活性的研究[J]. 中国食品学报, 2016, 16(12): 23-29.
