植物淀粉对乳酸片球菌微胶囊冻干保护及储存性能的影响

doi:10.16431/j.cnki.1671-7236.2024.10.040

摘要/Abstract

摘要： 【目的】研究马铃薯淀粉和玉米淀粉对冷冻干燥过程中微胶囊乳酸片球菌活性的保护作用和对乳酸片球菌耐酸耐胆盐性、储藏稳定性的影响，为抗性淀粉的开发利用及益生菌保护技术开拓思路和提供理论依据。【方法】选用3.0%海藻酸钠、0.8%壳聚糖制备3组微胶囊：对照组、试验1组(T1)和试验2组(T2)。对照组以2%甘油作为冻干保护剂，T1组以2%甘油和10%马铃薯淀粉作为冻干保护剂，T2组以2%甘油和10%玉米淀粉作为冻干保护剂。分别对微胶囊的外貌形态、机械强度、包埋率、载菌量、体外人工胃肠道耐受性及储藏性能进行评价。【结果】 ①T1和T2组微胶囊包埋率均显著高于对照组(P＜0.05)。②T1和T2组微胶囊冻干保护率达到70%以上，均显著高于对照组(P＜0.05)，其中T2组冻干保护率较高，为77.66%。③T1与T2组微胶囊的粒径、机械强度、载菌量均显著高于对照组(P＜0.05)。④体外人工胃肠液耐受性试验结果显示，T1和T2组微胶囊在含胃蛋白酶的模拟胃液中的存活率均高于对照组，2 h时差异显著(P＜0.05)；在模拟肠液中，3组乳酸片球菌微胶囊均在10 min后开始迅速释放，1 h时，基本崩解完全；T1与T2组乳酸片球菌微胶囊在胆盐环境下较稳定，4 h时乳酸片球菌存活率显著高于对照组(P＜0.05)；⑤3组微胶囊25 ℃储藏8周后，T1和T2组微胶囊乳酸片球菌存活率显著高于对照组(P＜0.05)，其中T2组微胶囊乳酸片球菌存活率较高，为52.29%；－20 ℃储存8周后，T1和T2组微胶囊乳酸片球菌存活率均高于对照组，但差异不显著(P＞0.05)。【结论】通过微囊化技术和添加植物淀粉作为保护剂的措施可提高乳酸片球菌微胶囊冻干保护率及储藏性能，其中玉米淀粉对乳酸片球菌微胶囊冻干保护及储存性能更好。本研究结果可为开发海藻酸钠、壳聚糖为壁材的乳酸片球菌微胶囊研究提供参考。

关键词: 乳酸片球菌; 马铃薯淀粉; 玉米淀粉; 微胶囊

Abstract: 【Objective】 This study was aimed to investigated the protective effect of potato starch and corn starch on the activity of microcapsulated Pediococcus acidilactici in the process of freeze-drying,as well as the effect on the acid and bile salts resistance of Pediococcus acidilactici and the storage stability of Pediococcus acidilactici,so as to explore ideas and provide theoretical basis for the development and utilization of resistant starch and probiotic protection technology. 【Method】 Three groups of microcapsules were prepared with 3.0% sodium alginate and 0.8%chitosan concentration:Control group,test group 1 (T1) and test group 2 (T2).In control group, 2% glycerol was used as lyophilizing agent,in group T1, 2% glycerol and 10% potato starch was used as lyophilizing agent,and in T2 group, 2% glycerol and 10% corn starch was used as lyophilizing agent.The morphology,mechanical strength,embedding rate,bacterial load,tolerance of artificial gastrointestinal tract in vitro and storage performance of the microcapsules were evaluated.【Result】 ①The embedding rate of microcapsule in groups T1 and T2 was significantly higher than that in control group (P＜0.05).②The lyophilization protection rate of groups T1 and T2 reached more than 70%,which were both significantly higher than that of control group (P＜0.05),and with a higher lyophilization protection rate of 77.66% in group T2.③The particle size,mechanical strength,bacterial load of microcapsules in groups T1 and T2 were significantly higher than those in control group (P＜0.05).④The in vitro tolerance test of artificial gastrointestinal fluids revealed that the survival rate of groups T1 and T2 in simulated gastric fluid containing pepsin was higher than that of control group,and the difference was significant after 2 h (P＜0.05).The Pediococcus acidilactici microcapsules in three groups started to release rapidly in simulated intestinal fluid after 10 min,and at 1 h,they were basically completely disintegrated.Pediococcus acidilactici microcapsules in groups T1 and T2 were stable in bile salt environment.The survival rate of Pediococcus acidilactici in groups T1 and T2 in bile salt environment was significantly higher than that of control group (P＜0.05) after 4 h.⑤Three groups of microcapsules were stored at 25 ℃ for 8 weeks,the survival rate of Pediococcus acidilactici in groups T1 and T2 was significantly higher than that in control group (P＜0.05),and the survival rate of Pediococcus acidilactici in group T2 was higher (52.29%).After 8 weeks of storage at －20 ℃,the survival rate of Pediococcus acidilactici in groups T1 and T2 was higher than that in control group,but the difference was not significant (P＞0.05).【Conclusion】 In summary,through the measures of microencapsulation technology and the addition of plant starch as a protective agent,the freeze-drying protection rate and storage performance of Pediococcus acidilactici microcapsules could be improved,among which corn starch had higher freeze-drying protection and storage performance of Pediococcus acidilactici microcapsules.The results could provide reference for the development of Pediococcus acidilactici microcapsules with sodium alginate and chitosan as wall materials.

Key words: Pediococcus acidilactici; potato starch; corn starch; microcapsules

中图分类号:

S816.7

张馨钰, 钟明, 崔莲花, 于小洋, 徐紫嫣, 李光春. 植物淀粉对乳酸片球菌微胶囊冻干保护及储存性能的影响[J]. 中国畜牧兽医, 2024, 51(10): 4596-4605.

ZHANG Xinyu, ZHONG Ming, CUI Lianhua, YU Xiaoyang, XU Ziyan, LI Guangchun. Effect of Plant Starch on Freeze-drying Protection and Storage Performance of Pediococcus acidilactici Microcapsules[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(10): 4596-4605.

参考文献

[1] 周莉,张立攀,关炳峰，等.嗜酸乳杆菌冻干微胶囊的制备及耐受性研究[J].食品科技,2021,46(7):43-47.ZHOU L,ZHANG L P,GUAN B F,et al.Preparation and tolerance of freeze-dried microcapsules of Lactobacillus acidophilus[J].Food Science and Technology,2021,46(7):43-47.(in Chinese)
[2] NABAVI-RAD A,SADEGHI A,ASADZADEH AGHDAEI H,et al.The double-edged sword of probiotic supplementation on gut microbiota structure in Helicobacter pylori management[J]. Gut Microbes,2022,14(1):2108655.
[3] 田文静,王俊国,宋娇娇，等.适宜冷冻干燥保护剂提高植物乳杆菌LIP-1微胶囊性能[J].农业工程学报,2015,31(21):285-294.TIAN W J,WANG J G,SONG J J,et al.Proper cryoprotectants improving properties of L.plantarum LIP-1 microcapsules[J].Transactions of the Chinese Society of Agricultural Engineering,2015,31(21):285-294.(in Chinese)
[4] 张琳,贠婷婷,綦文涛，等.乳化凝胶化法微囊化益生菌抗胁迫作用研究[J].动物营养学报,2015,27(11):3636-3642.ZHANG L,YUN T T,QI W T,et al.A study on the resistance of microencapsulated probiotics by the emulsion and internal gelation method[J]. Chinese Journal of Animal Nutrition,2015,27(11):3636-3642.(in Chinese)
[5] LIU J,LIU F,REN T,et al.Fabrication of fish gelatin/sodium alginate double network gels for encapsulation of probiotics[J].Journal of the Science of Food and Agriculture,2021,101(10):4398-4408.
[6] 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].Critical Reviews in Food Science and Nutrition,2019,59(17):2863-2878.
[7] 李昱辉,刘湘霞,黄才创.抗性淀粉的制备、来源、健康收益及其展望[J].轻工科技,2023,39(3):17-20.LI Y H,LIU X X,HUANG C C.Preparation,source,health benefits and prospect of resistant starch[J].Light Industry Science and Technology,2023,39(3):17-20.(in Chinese)
[8] 周中凯,王俊轩,李莹.青香蕉淀粉对乳酸菌在冻干和贮存过程中活性的保护效果[J].食品科学,2016,37(15):144-148.ZHOU Z K,WANG J X,LI Y.Protective effect of unripe banana starch on viability of Lactobacillus plantarum during freeze-drying and storage[J].Food Science,2016,37(15):144-148.(in Chinese)
[9] VELLOSO C C V,LOPES M M,BADINO A C,et al.Exploring the roles of starch for microbial encapsulation through a systemic mapping review[J].Carbohydrate Polymers,2023,306:120574.
[10] 陈胜杰,高翔,袁戎宇.真空冷冻干燥法制备益生菌粉的冻干保护剂配方优化[J].食品工业科技,2021,42(1):182-187.CHEN S J,GAO X,YUAN R Y.Formulation optimization of freeze-drying protectant for probiotic powder by vacuum freeze-drying[J].Science and Technology of Food Industry,2021,42(1):182-187.(in Chinese)
[11] CUI L H,YAN C G,LI H S,et al.A new method of producing a natural antibacterial peptide by encapsulated probiotics internalized with inulin nanoparticles as prebiotics[J].Journal of Microbiology and Biotechnology,2018,28(4):510-519.
[12] ZHOU T,KANG W,HAN Y,et al.Combinedly increased viability of Lactiplantibacillus plantarum grx16 by co-encapsulation of cryoprotectants and porous starch within calcium alginate capsules[J].International Journal of Food Science & Technology,2023,58(10):5291-5298.
[13] 崔乃元,高青山,严昌国，等.果胶/壳聚糖合生元微胶囊制备及其体外人工胃肠道耐受性和储藏性能研究[J].中国畜牧兽医,2020,47(9):2799-2807.CUI N Y,GAO Q S,YAN C G,et al.Preparation of pectin/chitosan synbiotic microcapsules and its study on simulated gastrointestinal environments and storage stability[J].China Animal Husbandry & Veterinary Medicine,2020,47(9):2799-2807.(in Chinese)
[14] 徐紫嫣,姜洋,刘学涛，等.果胶/壳聚糖姜黄素微胶囊制备及其体外人工胃肠道耐受性研究[J].中国畜牧兽医,2023,50(6):2312-2320.XU Z Y,JIANG Y,LIU X T,et al.Preparation of pectin/chitosan curcumin microcapsules and its study on simulated gastrointestinal environments[J].China Animal Husbandry & Veterinary Medicine,2023,50(6):2312-2320.(in Chinese)
[15] SAEED F,AFZAAL M,AHMAD A,et al.Enhanced viability of microencapsulated lyophilized probiotics under in vitro simulated gastrointestinal conditions[J].Journal of Food Processing and Preservation,2022,46(5):e16543.
[16] LEE Y J,JI Y R,LEE S,et al.Microencapsulation of probiotic Lactobacillus acidophilus KBL409 by extrusion technology to enhance survival under simulated intestinal and freeze-drying conditions[J]. Journal of Microbiology and Biotechnology,2019,29(5):721-730.
[17] 秦梓椋,陈少俊,何鑫淼，等.猪传染性胃肠炎病毒微胶囊疫苗的制备及其免疫效果评价[J].中国畜牧兽医,2023,50(9):3790-3798.QIN Z L,CHEN S J,HE X M,et al.Preparation of microencapsulated vaccine against Transmissible gastroenteritis virus and its immunological efficacy in piglets[J].China Animal Husbandry & Veterinary Medicine,2023,50(9):3790-3798.(in Chinese)
[18] KHLIBSUWAN R,TANSENA W,PONGJANYAKUI T.Modification of alginate beads using gelatinized and ungelatinized arrowroot (Tacca leontopetaloides L.Kuntze) starch for drug delivery[J].International Journal of Biological Macromolecules,2018,118:683-692.
[19] BENAVENT-GIL Y,RODRIGO D,ROSELL C M.Thermal stabilization of probiotics by adsorption onto porous starches[J].Carbohydrate Polymers,2018,197:558-564.
[20] HOYOS-LEYVA J D,BELLO-PEREZ L A,ALVAREZ-RAMIREZ J,et al.Microencapsulation using starch as wall material:A review[J].Food Reviews International,2018,34(2):148-161.
[21] QI X,TESTER R F.Starch granules as active guest molecules or microorganism delivery systems[J].Food Chemistry,2019,271:182-186.
[22] WOLKERS W F,PLDENHOF H.Principles underlying cryopreservation and freeze-drying of cells and tissues[J]. Cryopreservation and Freeze-drying Protocols,2021，2180:3-25.
[23] GUO Y,QIAO D,ZHAO S,et al.Starch-based materials encapsulating food ingredients:Recent advances in fabrication methods and applications[J].Carbohydrate Polymers,2021,270:118358.
[24] YUAN Y,YIN M,CHEN L,et al.Effect of calcium ions on the freeze-drying survival of probiotic encapsulated in sodium alginate[J].Food Hydrocolloids,2022,130:107668.
[25] LOZANO-VAZQUEZ G,LOBATO-CALLEROS C,ESCALONA-BUENDIA H,et al.Effect of the weight ratio of alginate-modified tapioca starch on the physicochemical properties and release kinetics of chlorogenic acid containing beads[J].Food Hydrocolloids,2015,48:301-311.
[26] CUI C,JIANG H,GUAN M,et al.Characterization and in vitro digestibility of potato starch encapsulated in calcium alginate beads[J].Food Hydrocolloids,2022,126:107458.
[27] 程龙,郝艳玲,宋小双.壳聚糖对玉米淀粉可食膜性能的影响[J].中国塑料,2021,35(4):35-41.CHENG L,HAO Y L,SONG X S.Effect of chitosan on properties of edible corn starch films[J].China Plastics,2021,35(4):35-41.(in Chinese)
[28] 毛金超,隋振全,徐桂云，等.天然生物质材料的制备、性质与应用(Ⅱ)——pH敏感型功能海洋多糖：海藻酸钠[J].日用化学工业,2022,52(2):124-133.MAO J C,SUI Z Q,XU G Y,et al.Preparation,properties and applications of natural biomass materials (Ⅱ) pH-sensitive functional marine polysaccharide:Sodium alginate[J].China Surfactant Detergent & Cosmetics,2022,52(2):124-133.(in Chinese)
[29] 高洪霞.益生菌递送体系的构建及其胃肠环境保护[D].南昌：南昌大学,2023.GAO H X.Construction and gastrointestinal environmental protection of probiotics delivery system [D].Nanchang:Nanchang University,2023.(in Chinese)
[30] HEIDEBACH T,FORST P,KULOZIK U.Influence of casein-based microencapsulation on freeze-drying and storage of probiotic cells[J].Journal of Food Engineering,2010,98(3):309-316.
[31] MUHAMMAD Z,RAMZAN R,ZHANG R,et al.Resistant starch-based edible coating composites for spray-dried microencapsulation of Lactobacillus acidophilus,comparative assessment of thermal protection,in vitro digestion and physicochemical characteristics[J].Coatings,2021,11(5):587.