食品与发酵工业 >

2025 , Vol. 51 >Issue 13: 62 - 68

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.040466

研究报告

真空冷冻干燥过程中长双歧杆菌长亚种细胞损伤的机制

  • 冯海红 ,
  • 王然 ,
  • 葛绍阳 ,
  • 张永祥 ,
  • 赵亮 ,
  • 李晓霞 ,
  • 姚凯 ,
  • 桑跃
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  • 1(中国农业大学 营养与健康系,北京,100193)
    2(北京和益源生物技术有限公司,北京,100089)
    3(内蒙古兰格格乳业有限公司,内蒙古 乌兰察布,012000)
第一作者: 硕士,中级工程师(桑跃助理研究员为通信作者,E-mail:sangyue2013@126.com)

收稿日期: 2024-07-11

  修回日期: 2024-09-26

  网络出版日期: 2025-08-04

基金资助

内蒙古自治区“揭榜挂帅”项目(2022JBGS0002);乌兰察布市科技计划项目(2023PT402)

收起

Mechanism of cell damage of Bifidobacterium longum subsp. longum during freeze-drying process

  • FENG Haihong ,
  • WANG Ran ,
  • GE Shaoyang ,
  • ZHANG Yongxiang ,
  • ZHAO Liang ,
  • LI Xiaoxia ,
  • YAO Kai ,
  • SANG Yue
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  • 1(Department of Nutrition and Health, China Agricultural University, Beijing 100193, China)
    2(Beijing Heyiyuan Biotechnology Company Limited, Beijing 100089, China)
    3(Inner Mongolia Langege Dairy Co.Ltd., Ulanchap 012000, China)

Received date: 2024-07-11

  Revised date: 2024-09-26

  Online published: 2025-08-04

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摘要

为明确真空冷冻干燥过程中长双歧杆菌长亚种细胞损伤的主要阶段和损伤原因。该研究测定了长双歧杆菌长亚种BB68S在真空冷冻干燥不同阶段,包括预冻阶段、升华干燥阶段和解吸干燥阶段菌体细胞的存活性、水分变化、细胞壁完整性和细胞膜通透性等特征。结果表明,预冻阶段自由水在低温下形成冰晶,挤压细胞,导致细胞壁肽聚糖结构受损,降低了细胞壁完整度,细胞存活性下降14.1%;升华干燥阶段水分大量散失,导致细胞膜磷脂双分子层脱水后结构改变,通透性增加,致使乳酸脱氢酶、β-半乳糖苷酶等胞内代谢关键酶渗漏到细胞外,细胞活性剧烈降低49.7%;解析干燥阶段水分和细胞存活性变化均较小。因此,BB68S冻干过程中菌体细胞损伤的主要阶段为预冻阶段和升华干燥阶段,主要是由于水分变化引起细胞壁和细胞膜结构损伤,降低了其功能性,最终导致细胞活性下降。该研究解析了长双歧杆菌长亚种在冻干过程中细胞损伤的机制,为控制真空冷冻干燥工艺提高菌株冻干存活性提供了理论依据。

关键词： 长双歧杆菌长亚种; 真空冷冻干燥; 细胞壁; 细胞膜; 关键代谢酶

本文引用格式

冯海红 , 王然 , 葛绍阳 , 张永祥 , 赵亮 , 李晓霞 , 姚凯 , 桑跃 . 真空冷冻干燥过程中长双歧杆菌长亚种细胞损伤的机制[J]. 食品与发酵工业, 2025 , 51(13) : 62 -68 . DOI: 10.13995/j.cnki.11-1802/ts.040466

Abstract

This study aimed to clarify the primary stages and caused of cell damage in Bifidobacterium longum subsp. longum during vacuum freeze-drying.This study assessed the cell viability, water content, cell wall integrity, and cell membrane permeability of B.longum subsp. longum BB68S at various stages of the vacuum freeze-drying process, including the pre-freezing stage, sublimation drying stage, and desorption drying stage.Results revealed that, during the pre-freezing stage, the formation of ice crystals from free water at low temperatures caused cell extrusion.This process resulted in damage to the peptidoglycan structure of the cell wall, leading to reduced cell wall integrity and a 14.1% decrease in cell viability.In the sublimation drying stage, significant water loss induced structural changes in the phospholipid bilayer of the cell membrane following dehydration, resulting in increased permeability.This alteration led to the leakage of key intracellular metabolic enzymes such as lactate dehydrogenase and β-galactosidase into the external environment, causing a drastic 49.7% decrease in cell activity.Water content and cell viability changes were minimal during the desorption drying stage.Therefore, the primary stages of cellular damage during B.longum subsp. longum BB68S lyophilization occurred during pre-freezing and sublimation drying stages due to moisture-related compromises on both cell wall and membrane structures, ultimately reducing functionality and leading to decreased cellular activity.This study analyzed mechanisms of B.longum subsp. longum cellular damage during freeze-drying processes and provided a theoretical basis for optimizing vacuum freeze-drying procedures to enhance strain survivability.

Key words： Bifidobacterium longum subsp. longum; vacuum freeze-drying; cell wall; cell membrane; key metabolic enzymes

参考文献

[1] CHEN Z C, E J J, MA R Z, et al.The effect of aspartic acid on the freeze-drying survival rate of Lactobacillus plantarum LIP-1 and its inherent mechanism[J].LWT, 2022, 155:112929.
[2] E J J, ZHANG J Y, MA R Z, et al.Study of the internal mechanism of L-glutamate for improving the survival rate of Lactiplantibacillus plantarum LIP-1 after freeze-drying[J].Innovative Food Science & Emerging Technologies, 2023, 84:103253.
[3] YANG Y X, WANG R X, CHEN Z C, et al.Effects of exogenous amino acids on the freeze-drying survival rate of Lactiplantibacillus plantarum LIP-1[J].Drying Technology, 2024, 42(4):650-660.
[4] SCHWAB C, VOGEL R, GÄNZLE M G.Influence of oligosaccharides on the viability and membrane properties of Lactobacillus reuteri TMW1.106 during freeze-drying[J].Cryobiology, 2007, 55(2):108-114.
[5] LIU X T, HOU C L, ZHANG J, et al.Fermentation conditions influence the fatty acid composition of the membranes of Lactobacillus reuteri I5007 and its survival following freeze-drying[J].Letters in Applied Microbiology, 2014, 59(4):398-403.
[6] KANDIL S, EL SODA M.Influence of freezing and freeze drying on intracellular enzymatic activity and autolytic properties of some lactic acid bacterial strains[J].Advances in Microbiology, 2015, 5(6):371-382.
[7] CHEN H, CHEN S W, LI C N, et al.Response surface optimization of lyoprotectant for Lactobacillus bulgaricus during vacuum freeze-drying[J].Preparative Biochemistry & Biotechnology, 2015, 45(5):463-475.
[8] JIANG G, WANG J, ZHAO D D, et al.Molecular mechanism of the cytosine CRISPR base editing process and the roles of translesion DNA polymerases[J].ACS Synthetic Biology, 2021, 10(12):3353-3358.
[9] 李明慧, 尚一娜, 霍麒文, 等.真空冷冻干燥对乳酸菌损伤机制的研究进展[J].食品科学, 2018, 39(19):273-279.
LI M H, SHANG Y N, HUO Q W, et al.Recent progress in understanding injury mechanisms of lactic acid bacteria during freeze-drying[J].Food Science, 2018, 39(19):273-279.
[10] 刘开文, 马雯, 金刚.真空冷冻干燥提高乳酸菌存活率及延长贮藏期的研究进展[J].食品科学, 2024, 45(2):325-333.
LIU K W, MA W, JIN G.Research progress on improving the survival rate and extending the storage period of lactic acid bacteria through vacuum freeze-drying[J].Food Science, 2024, 45(2):325-333.
[11] 丁圣, 蒋菁莉, 刘松玲, 等.长双歧杆菌BBMN68对便秘模型小鼠的通便作用[J].食品科学, 2011, 32(3):195-198.
DING S, JIANG J L, LIU S L, et al.Effect of biofidobactria strain on defecating function in constipation mice[J].Food Science, 2011, 32(3):195-198.
[12] YUN B, SONG M Y, PARK D J, et al.Beneficial effect of Bifidobacterium longum ATCC 15707 on survival rate of Clostridium difficile infection in mice[J].Korean Journal for Food Science of Animal Resources, 2017, 37(3):368-375.
[13] 韦云路, 刘义, 王瑶, 等.3株益生菌体外降胆固醇能力及体内降血脂效果评价[J].食品科学, 2017, 38(23):129-134.
WEI Y L, LIU Y, WANG Y, et al.Cholesterol-lowering ability in vitro and hypolipidemic effect in vivo of three probiotic strains[J].Food Science, 2017, 38(23):129-134.
[14] 吕锡斌, 何腊平, 张汝娇, 等.双歧杆菌生理功能研究进展[J].食品工业科技, 2013, 34(16):353-358.
LYU X B, HE L P, ZHANG R J, et al.Research progress in physiological functions of Bifidobacterium[J].Science and Technology of Food Industry, 2013, 34(16):353-358.
[15] 杨景, 罗绪刚, 任发政.长双歧杆菌BBMN68诱导的树突状细胞对小鼠牛乳β-乳球蛋白过敏的缓解作用[J].现代食品科技, 2016, 32(7):6-11.
YANG J, LUO X G, REN F Z.Reduction of bovine β-lactoglobulin allergenicity via Bifidobacterium longum BBMN68 modulated-dendritic cells in mice[J].Modern Food Science and Technology, 2016, 32(7):6-11.
[16] DUMONT F, MARECHAL P A, GERVAIS P.Cell size and water permeability as determining factors for cell viability after freezing at different cooling rates[J].Applied and Environmental Microbiology, 2004, 70(1):268-272.
[17] SENZ M, VAN LENGERICH B, BADER J, et al.Control of cell morphology of probiotic Lactobacillus acidophilus for enhanced cell stability during industrial processing[J].International Journal of Food Microbiology, 2015, 192:34-42.
[18] BRAVO-FERRADA B M, GONÇALVES S, SEMORILE L, et al.Study of surface damage on cell envelope assessed by AFM and flow cytometry of Lactobacillus plantarum exposed to ethanol and dehydration[J].Journal of Applied Microbiology, 2015, 118(6):1409-1417.
[19] 公丕民. 保加利亚乳杆菌喷雾干燥过程中损伤机制及保护方法研究[D].哈尔滨:哈尔滨工业大学, 2019.
GONG P M.Study on damage mechanism and protection method of Lactobacillus bulgaricus during spray drying[D].Harbin:Harbin Institute of Technology, 2019.
[20] SUNDARAMURTHI P, SURYANARAYANAN R.Calorimetry and complementary techniques to characterize frozen and freeze-dried systems[J].Advanced Drug Delivery Reviews, 2012, 64(5):384-395.
[21] SANTIVARANGKNA C, KULOZIK U, FOERST P.Effect of carbohydrates on the survival of Lactobacillus helveticus during vacuum drying[J].Letters in Applied Microbiology, 2006, 42(3):271-276.
[22] INAMURA S, FUKASE K, KUSUMOTO S.Synthetic study of peptidoglycan partial structures.Synthesis of tetrasaccharide and octasaccharide fragments[J].Tetrahedron Letters, 2001, 42(43):7613-7616.
[23] 刘佳, 蔡开聪.β(1→4)糖苷键振动光谱与纤维二糖结构的相关性研究[J].光谱学与光谱分析, 2016, 36(S1):60-61.
LIU J, CAI K C.The correlation between the vibrational spectrum of β(1→4)glycosidic linkage and the structure of cellobiose[J].Spectroscopy and Spectral Analysis, 2016, 36(S1):60-61.
[24] LUO C, LIU Z Q, MI S, et al.Quantitative investigation on the effects of ice crystal size on freeze-drying:The primary drying step[J].Drying Technology, 2022, 40(2):446-458.
[25] SANTIVARANGKNA C, WENNING M, FOERST P, et al.Damage of cell envelope of Lactobacillus helveticus during vacuum drying[J].Journal of Applied Microbiology, 2007, 102(3):748-756.
[26] LOUESDON S, CHARLOT-ROUGÉ S, JUILLARD V, et al.Osmotic stress affects the stability of freeze-dried Lactobacillus buchneri R1102 as a result of intracellular betaine accumulation and membrane characteristics[J].Journal of Applied Microbiology, 2014, 117(1):196-207.
[27] SANTIVARANGKNA C, KULOZIK U, FOERST P.Inactivation mechanisms of lactic acid starter cultures preserved by drying processes[J].Journal of Applied Microbiology, 2008, 105(1):1-13.
[28] BASHOLLI-SALIHU M, MUELLER M, SALAR-BEHZADI S, et al.Effect of lyoprotectants on β-glucosidase activity and viability of Bifidobacterium infantis after freeze-drying and storage in milk and low pH juices[J].LWT - Food Science and Technology, 2014, 57(1):276-282.
[29] JOFRÉ A, AYMERICH T, GARRIGA M.Impact of different cryoprotectants on the survival of freeze-dried Lactobacillus rhamnosus and Lactobacillus casei/paracasei during long-term storage[J].Beneficial Microbes, 2015, 6(3):381-386.
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