工业化生产冻干前不同阶段双歧杆菌死亡规律的模拟分析
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摘要
为降低双歧杆菌生产过程中的死亡损失,模拟分析其工业化生产冻干前不同阶段的死亡规律。首先测定两歧双歧杆菌CCFM16、青春双歧杆菌CCFM8630、短双歧杆菌CCFM683和长双歧杆菌L6发酵液暴露于空气中不同时间菌株的存活率,然后比较不同温度、时间和离心力离心后菌体的存活率,最后测定菌泥、及菌泥与保护剂混合后的菌悬液暴露至室温环境不同时间的存活率。发现发酵液空气暴露3 h后4株菌的活性损失约为20%; 4℃低温离心和短离心时间更有利于菌体活性的保持,离心力影响不显著;菌泥暴露室温环境中1 h死亡接近或超过10%;与保护剂混合的菌悬液暴露室温2 h内双歧杆菌死亡率接近或超过10%,暴露3 h后4株菌的死亡接近或超过20%。通过在保护剂中添加D-异抗坏血酸钠作为抗氧化剂可显著提高双歧杆菌的存活率。该研究阐明了生产阶段影响双歧杆菌活性的关键因素,为提高双歧杆菌的生产效率具有指导意义。
In order to reduce the death rate of Bifidobacterium during industrial preparation,its death patterns at different stages before freeze-drying were experimentally analyzed. The survival rates of Bifidobacterium bifidum CCFM16,Bifidobacterium adolescentis CCFM8630,Bifidobacterium brevis CCFM683 and Bifidobacterium longum L6 were determined under different processing conditions. It was found that the survival rate of the four strains decreased about 20% after exposing the fermentation broth to air for 3 h. Quick centrifugation at 4 °C was beneficial for lowering the death rate,while centrifugation force in a certain range had no significant effect. Survival rate of the pellet decreased about 10% after exposing to air for 1 h. The same decrease was found while the mixture of lyoprotectant and bacterial suspension exposed to air for 2 h,and its survival rate decreased about 20% in a 3 h exposure. Additionally,their survival rates were significantly improved by D-isoascorbate. This study clarified the key factors that influence the survival rate of Bifidobacterium during preparation and therefore has guiding significance for improving its production efficiency.
In order to reduce the death rate of Bifidobacterium during industrial preparation,its death patterns at different stages before freeze-drying were experimentally analyzed. The survival rates of Bifidobacterium bifidum CCFM16,Bifidobacterium adolescentis CCFM8630,Bifidobacterium brevis CCFM683 and Bifidobacterium longum L6 were determined under different processing conditions. It was found that the survival rate of the four strains decreased about 20% after exposing the fermentation broth to air for 3 h. Quick centrifugation at 4 °C was beneficial for lowering the death rate,while centrifugation force in a certain range had no significant effect. Survival rate of the pellet decreased about 10% after exposing to air for 1 h. The same decrease was found while the mixture of lyoprotectant and bacterial suspension exposed to air for 2 h,and its survival rate decreased about 20% in a 3 h exposure. Additionally,their survival rates were significantly improved by D-isoascorbate. This study clarified the key factors that influence the survival rate of Bifidobacterium during preparation and therefore has guiding significance for improving its production efficiency.
引文
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[16]任香芸,何志刚,李维新,等.直投式发酵剂制备工艺对乳酸菌存活率的影响[J].食品科学技术学报,2017,35(6):36-41.
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[19]TRIPATHI M K,GIRI S K.Probiotic functional foods:Survival of probiotics during processing and storage[J].Journal of Functional Foods,2014,9:225-241.
[2]SISTEK D,KELLY R,WICKENS K,et al.Is the effec of probiotics on atopic dermatitis confined to food sensitized children[J].Clinical and Experimental Allergy,2006,36(5):629-633.
[3]胡会萍.益生菌及其在功能食品中的应用[J].食品研究与开发,2007,28(2):173-175.
[4]ALLEN A P,CLARKE G,CRYAN J F,et al.Bifidobacterium infantis 35624 and other probiotics in the management of irritable bowel syndrome.Strain specificity,symptoms,and mechanisms[J].Current Medical Research&Opinion,2017,33(7):1 349-1 351.
[5]STAUDACHER H M,LOMER M C,FARQUHARSON FM,et al.A diet low in FODMAPs reduces symptoms in patients with irritable bowel syndrome and a probiotic restores Bifidobacterium species:A randomized controlled trial[J].Gastroenterology,2017,153(4):936-947.
[6]LAU A S Y,YANAGISAWA N,HOR Y Y,et al.Bifidobacterium longum BB536 alleviated upper respiratory illnesses and modulated gut microbiota profiles in Malaysian pre-school children[J].Beneficial Microbes,2018,9(1):61-70.
[7]MOYA-PREZ A,PEREZ-VILLALBA A,BENTEZ-PEZ A,et al.Bifidobacterium CECT 7765 modulates early stress-induced immune,neuroendocrine and behavioral alterations in mice[J].Brain,Behavior,and Immunity,2017,65:43-56.
[8]万红兵,田洪涛,马晓燕,等.直投式酸奶发酵剂制备过程中乳酸菌离心分离条件的研究[J].食品工业科技,2006,27(11):69-71;75.
[9]MORGAN C A,HERMAN N,WHITE P A,et al.Preservation of micro-organisms by drying;A review[J].Journal of Microbiological Methods,2006,66(2):183-193.
[10]CORCORAN B M,ROSS R P,FITZGERALD G F,et al.Comparative survival of probiotic lactobacilli spraydried in the presence of prebiotic substances[J].Journal of Applied Microbiology,2004,96(5):1 024-1 039.
[11]ZHANG G,FAN M,LI Y,et al.Effect of growth phase,protective agents,rehydration media and stress pretreatments on viability of Oenococcus oeni subjected to freezedrying[J].African Journal of Microbiology Research,2012,6(7):1 478-1 484.
[12]MENG X C,STANTON C,FITZGERALD G F,et al.Anhydrobiotics:The challenges of drying probiotic cultures[J].Food Chemistry,2008,106(4):1 406-1 416.
[13]李青青.耐氧性双歧杆菌的筛选及其生理特性与应用研究[D].杭州:浙江大学,2010.
[14]柳翰凌,康小红.双歧杆菌厌氧机制的初步探讨[J].食品科技,2008,33(3):15-17.
[15]PETERSON B W,SHARMA P K,VAN DER MEI H C,et al.Bacterial cell surface damage due to centrifugal compaction[J].Applied and Environmental Microbiology,2012,78(1):120-125.
[16]任香芸,何志刚,李维新,等.直投式发酵剂制备工艺对乳酸菌存活率的影响[J].食品科学技术学报,2017,35(6):36-41.
[17]白少峰,陈华海,王欣,等.双歧杆菌胞外多糖研究进展[J].中国微生态学杂志,2017,29(10):1 207-1 211;1 218.
[18]CUI S,HANG F,LIU X,et al.Effect of acids produced from carbohydrate metabolism in cryoprotectants on the viability of freeze-dried Lactobacillus and prediction of optimal initial cell concentration[J].Journal of Bioscience and Bioengineering,2018,125(5):513-518.
[19]TRIPATHI M K,GIRI S K.Probiotic functional foods:Survival of probiotics during processing and storage[J].Journal of Functional Foods,2014,9:225-241.