研究报告

黄芪多糖对嗜酸乳杆菌CICC 22162冷冻干燥保护效果的研究

  • 杜建明 ,
  • 张珍 ,
  • 尚琪 ,
  • 王学瑞 ,
  • 史建莉 ,
  • 宋乐 ,
  • 谢鹏 ,
  • 蔡毓涵
展开
  • (甘肃农业大学 食品科学与工程学院,甘肃 兰州,730000)
第一作者:硕士研究生(张珍教授为通信作者,E-mail:zhangzhen@gsau.edu.cn)

收稿日期: 2025-03-21

  修回日期: 2025-07-03

  网络出版日期: 2026-03-25

基金资助

甘肃省农业农村厅科技支撑项目(KJZC-2025-27);中国乳业协会利乐专项基金(CDIAKCJJ-TP-2024-008);甘肃省产业支撑项目(2023CYZC-43);甘肃省现代寒旱特色农业药产业技术体系(GSARS06-06)

Cryoprotective effects of astragalus polysaccharides on freeze-drying survival of Lactobacillus acidophilus CICC 22162

  • DU Jianming ,
  • ZHANG Zhen ,
  • SHANG Qi ,
  • WANG Xuerui ,
  • SHI Jianli ,
  • SONG Le ,
  • XIE Peng ,
  • CAI Yuhan
Expand
  • (College of Food Science and Engineering, Gansu Agricultural University, Lanzhou 730000, China)

Received date: 2025-03-21

  Revised date: 2025-07-03

  Online published: 2026-03-25

摘要

黄芪多糖具有多种生理活性功能,其高效利用具有重要意义。该研究以提取纯化的黄芪多糖为原料,并与其他冻干保护剂复配,测定其对嗜酸乳杆菌CICC 22162冷冻干燥的保护效果。结果表明,黄芪多糖组分为64.38%葡萄糖、19.81%半乳糖和15.81%阿拉伯糖,复合保护剂使嗜酸乳杆菌存活率提升至(85.02±1.25)%。扫描电镜显示菌体形态更均匀,相较于传统保护剂,复合保护剂的碱性磷酸酶活性(3 450.61 U/g)降低15.5%,乳酸脱氢酶泄漏减少38.7%,溶菌酶敏感性实验中活菌数降幅减少20.6%,NaCl胁迫下膜损伤降低37.2%,组蛋白外泄量减少68%。证明黄芪多糖能更好地维持细胞壁和细胞膜的完整性,减少细胞内容物的泄漏,抵抗外界不利的环境变化。在模拟胃肠消化中菌株存活率达81.3%,较传统保护剂提升33.9%,4 ℃贮藏120 d后活菌数仍有9.18 lg CFU/mL,制备的冻干菌粉兼具高存活率、强胃肠耐受性和稳定性,研究结果为益生菌制剂提供了创新保护方案,在功能性食品和肠道微生态调节领域具有重要应用价值。

本文引用格式

杜建明 , 张珍 , 尚琪 , 王学瑞 , 史建莉 , 宋乐 , 谢鹏 , 蔡毓涵 . 黄芪多糖对嗜酸乳杆菌CICC 22162冷冻干燥保护效果的研究[J]. 食品与发酵工业, 2026 , 52(5) : 139 -147 . DOI: 10.13995/j.cnki.11-1802/ts.042794

Abstract

Astragalus polysaccharide (APS) exhibits multiple physiological activities, making its efficient development and utilization highly significant. This study investigated the cryoprotective effects of purified APS (composed of 64.38% glucose, 19.81% galactose, and 15.81% arabinose) in combination with other cryoprotectants on Lactobacillus acidophilus CICC 22162 during freeze-drying.Results showed that the APS-based cryoprotectant significantly improved bacterial survival to (85.02±1.25)%.Scanning electron microscopy revealed more uniform cellular morphology, compared with conventional cryoprotectants, the APS-based formulation also resulted in a 15.5% lower than conventional protectants, lactate dehydrogenase leakage decreased by 38.7%, viability reduction in lysozyme sensitivity tests was 20.6% lower, NaCl-induced membrane damage was reduced by 37.2%, and histone phosphorylation leakage decreased by 68%.These findings proved APS better maintains cell wall/membrane integrity, reduced intracellular content leakage, and enhanced resistance to environmental stresses.The formulation achieved 81.3% survival in simulated gastrointestinal digestion (33.9% improvement over conventional protectants) and maintained 9.18 lg CFU/mL viability after 120 days at 4 ℃.The APS-based cryoprotectant enables production of freeze-dried probiotics with high viability, excellent gastrointestinal tolerance, and superior stability, representing an innovative protection strategy with significant application value in functional foods and gut microbiota regulation.

参考文献

[1] 姜辉, 顾胜龙, 张玉婷, 等.黄芪化学成分和药理作用研究进展[J].安徽中医药大学学报, 2020, 39(5):93-96.
JIANG H, GU S L, ZHANG Y T, et al.Research progress on chemical constituents and pharmacological effects of Astragalus membranaceus[J].Journal of Anhui University of Chinese Medicine, 2020, 39(5):93-96.
[2] LI C X, LIU Y, ZHANG Y Z, et al.Astragalus polysaccharide:A review of its immunomodulatory effect[J].Archives of Pharmacal Research, 2022, 45(6):367-389.
[3] 薛晨曦. 黄芪多糖合生元的制备与临床初步应用[D].长春:吉林大学, 2023.
XUE C X.Preparation and preliminary clinical application of astragalus polysaccharide synbiotics[D].Changchun:Jilin University, 2023.
[4] JING Y P, LI A P, LIU Z R, et al.Absorption of Codonopsis pilosula saponins by coexisting polysaccharides alleviates gut microbial dysbiosis with dextran sulfate sodium-induced colitis in model mice[J].BioMed Research International, 2018, 2018:1781036.
[5] WANG G Q, LUO L Y, DONG C, et al.Polysaccharides can improve the survival of Lactiplantibacillus plantarum subjected to freeze-drying[J].Journal of Dairy Science, 2021, 104(3):2606-2614.
[6] DONG Y S, LI M H, YUE X Q.Current research on probiotics and fermented products[J].Foods, 2024, 13(9):1406.
[7] MINJ J, CHANDRA P, PAUL C, et al.Bio-functional properties of probiotic Lactobacillus:Current applications and research perspectives[J].Critical Reviews in Food Science and Nutrition, 2021, 61(13):2207-2224.
[8] SHI Z H, LI X F, FAN X K, et al.The SlpX protein plays a crucial role in the intestinal juice tolerance of Lactobacillus acidophilus CICC6074[J].Food Bioscience, 2024, 59:103865.
[9] 周立光, 杨明喆, 冯会粉, 等.复合益生菌产品菌种鉴定及活菌定量检测方法[J].食品与发酵工业, 2022, 48(9):235-244.
ZHOU L G, YANG M Z, FENG H F, et al.Study on methods of species identification and quantitative detection of viable cells in composite probiotics products[J].Food and Fermentation Industries, 2022, 48(9):235-244.
[10] IBRAHIM I, AYARIGA J A, XU J H, et al.A comparative study of skimmed milk and cassava flour on the viability of freeze-dried lactic acid bacteria as starter cultures for yogurt fermentation[J].Foods, 2023, 12(6):1207.
[11] GAGNETEN M, PASSOT S, CENARD S, et al.Mechanistic study of the differences in lactic acid bacteria resistance to freeze- or spray-drying and storage[J].Applied Microbiology and Biotechnology, 2024, 108(1):361.
[12] LIU L, XIE S S, ZHU Y D, et al.Sodium carboxymethyl celluloses as a cryoprotectant for survival improvement of lactic acid bacterial strains subjected to freeze-drying[J].International Journal of Biological Macromolecules, 2024, 260:129468.
[13] STEFANELLO R F, NABESHIMA E H, IAMANAKA B T, et al.Survival and stability of Lactobacillus fermentum and Wickerhamomyces anomalus strains upon lyophilisation with different cryoprotectant agents[J].Food Research International, 2019, 115:90-94.
[14] WANG J, JIA J Y, SONG L, et al.Extraction, structure, and pharmacological activities of Astragalus polysaccharides[J].Applied Sciences, 2019, 9(1):122.
[15] 冉思琪. 黄芪多糖APS-1a的分离纯化、结构鉴定及促进肝细胞EPO分泌的研究[D].武汉:华中科技大学, 2020.
RAN S Q.Isolation, purification, structural characterization of polysaccharide APS-1a from Astragalus membranaceus and its activity of stimulating EPO secretion in HepG2 cell[D].Wuhan:Huazhong University of Science and Technology, 2020.
[16] 段腊梅. 苹果梨渣多糖结构表征及其与乳清蛋白相互作用研究[D].锦州:渤海大学, 2021.
DUAN L M.Structural characterization of polysaccharides from ‘Pingguo’ pear residue and their interactions with whey protein[D].Jinzhou:Bohai University, 2021.
[17] YANG W H, HAO J W, CHEN N D, et al.Development of a joint derivatization protocol for the unequivocal identification of the monosaccharide composition in four Dendrobium polysaccharides and free monosaccharide by GC-MS[J].Biomedical Chromatography, 2023, 37(12):e5743.
[18] MARTINS G N, GUERRERO SÁNCHEZ M, CARBONI A D, et al.Use of legume wastewater extracts on the storage stability of freeze-dried Lactiplantibacillus plantarum WCFS1[J].Food and Bioprocess Technology, 2025, 18(2):1707-1718.
[19] BROMLEY A, PERRY J.Survival of probiotic Lactobacillus spp.during kombucha fermentation[J].Current Developments in Nutrition, 2022, 6:507.
[20] ALIZADEH BEHBAHANI B, JOOYANDEH H, VASIEE A, et al.Evaluation of anti-yeast metabolites produced by Lactobacillus strains and their potential application as bio-preservatives in traditional yogurt drink[J].LWT, 2023, 188:115428.
[21] EL OIRDI S, LAKHLIFI T, FRABERGER V, et al.Probiotic features of three lactic acid bacteria strains isolated from foods and characterization of their bioactive molecules[J].Microbiology, 2023, 92(6):881-891.
[22] 唐美. 卷曲乳杆菌发酵培养基优化及其抑菌物质的初步分离纯化和鉴定[D].长沙:中南大学, 2022.
TANG M.Optimization of fermentation medium of Lactobacillus crispatus and preliminary isolation, purification and identification of their antibacterial substances[D].Changsha:Central South University, 2022.
[23] 刘敏敏, 席茂盛, 李中媛, 等.植物乳杆菌CGMCC8198冻干菌粉的制备工艺优化[J].食品研究与开发, 2020, 41(21):9-14.
LIU M M, XI M S, LI Z Y, et al.Preparation process optimization of Lactobacillus plantarum CGMCC8198 lyophilized powder[J].Food Research and Development, 2020, 41(21):9-14.
[24] GARCÍA-CEJA A, MANI-LÓPEZ E, PALOU E, et al.Viability during refrigerated storage in selected food products and during simulated gastrointestinal conditions of individual and combined lactobacilli encapsulated in alginate or alginate-chitosan[J].LWT-Food Science and Technology, 2015, 63(1):482-489.
[25] CHEN B Y, WANG X Y, LI P Z, et al.Exploring the protective effects of freeze-dried Lactobacillus rhamnosus under optimized cryoprotectants formulation[J].LWT, 2023, 173:114295.
[26] TANG Z J, HUANG G L.Extraction, structure, and activity of polysaccharide from Radix astragali[J].Biomedicine & Pharmacotherapy, 2022, 150:113015.
[27] LI K, LI X Q, LI G X, et al.Relationship between the structure and immune activity of components from the active polysaccharides APS-II of astragali Radix by enzymolysis of endo α-1, 4-glucanase[J].Frontiers in Pharmacology, 2022, 13:839635.
[28] 宋虹, 陈晨, 张尊琴, 等.大豆种皮多糖对副干酪乳杆菌6244生长及肠道黏附性的影响[J].中国食品学报, 2025, 25(1):142-150.
SONG H, CHEN C, ZHANG Z Q, et al.Effects of soybean hull polysaccharides on the growth and adhesion of Lactobacillus paracasei 6244[J].Journal of Chinese Institute of Food Science and Technology, 2025, 25(1):142-150.
[29] 李媛, 谢玉东, 王媛媛.银耳多糖对椰浆酸奶感官品质及益生菌生长的影响[J].食品安全导刊, 2024(29):113-118.
LI Y, XIE Y D, WANG Y Y.Effect of Tremella fuciformis polysaccharides on sensory quality and probiotic growth of coconut milk yogurt[J].China Food Safety Magazine, 2024(29):113-118.
[30] WANG H, SHAO L L, SUN Y Y, et al.Recovery mechanisms of ohmic heating-induced sublethally injured Staphylococcus aureus: Changes in cellular structure and applications in pasteurized milk[J].Food Control, 2025, 171:111086.
[31] TRIPATHI A, LIVERANI E, TSYGANKOV A Y, et al.Iron alters the cell wall composition and intracellular lactate to affect Candida albicans susceptibility to antifungals and host immune response[J].Journal of Biological Chemistry, 2020, 295(29):10032-10044.
[32] 张玉华, 孟一, 凌沛学, 等.海藻糖和透明质酸对冻干双歧杆菌细胞的保护作用[J].食品科学, 2010, 31(7):236-241.
ZHANG Y H, MENG Y, LING P X, et al.Protective mechanisms of trehalose and hyaluronic acid on lyophilized Bifidobacterium longum[J].Food Science, 2010, 31(7):236-241.
[33] MOAYYEDI M, ESKANDARI M H, RAD A H E, et al.Effect of drying methods (electrospraying, freeze drying and spray drying) on survival and viability of microencapsulated Lactobacillus rhamnosus ATCC 7469[J].Journal of Functional Foods, 2018, 40:391-399.
[34] GONG P M, SUN J L, LIN K, et al.Changes process in the cellular structures and constituents of Lactobacillus bulgaricus sp1.1 during spray drying[J].LWT, 2019, 102:30-36.
[35] 鄂晶晶. 培养条件对植物乳植杆菌LIP-1冷冻干燥存活率的影响及其内在机制的研究[D].呼和浩特:内蒙古农业大学,2022.
E J J.Effect of culture conditions on Lactiplantibacillus plantarum LIP-1 freeze-drying survival rate and the internal mechanism of action[D].Hohhot:Inner Mongolia Agricultural University, 2022.
[36] 王依朝, 樊丽, 王红霞, 等.γH2AX、53BP1及RAD51焦点用于分析DNA双链断裂损伤[J].生物学杂志, 2020, 37(1):16-19.
WANG Y Z, FAN L, WANG H X, et al.γH2AX, 53BP1 and RAD51 foci analysis for monitoring DNA double-strand breaks[J].Journal of Biology, 2020, 37(1):16-19.
[37] 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.
[38] WANG R X, E J J, YANG Y, et al.Trehalose enhances the resistance of Lactiplantibacillus plantarum LIP-1 to spray- and freeze-drying treatments by regulating amino acid metabolism[J].Food Bioscience, 2024, 60:104256.
[39] 邵匡振, 丁向萍, 张子恒, 等.中药益生元的研究进展[J].中国微生态学杂志, 2024, 36(7):838-842.
SHAO K Z, DING X P, ZHANG Z H, et al.Prebiotics in Chinese materia medica:Research progress[J].Chinese Journal of Microecology, 2024, 36(7):838-842.
[40] GE S T, HAN J R, SUN Q Y, et al.Optimization of cryoprotectants for improving the freeze-dried survival rate of potential probiotic Lactococcus lactis ZFM559 and evaluation of its storage stability[J].LWT, 2024, 198:116052.
文章导航

/

〈 〉