CN111436614A - 基于香菇可溶性膳食纤维的益生菌递送微胶囊及制备方法 - Google Patents
基于香菇可溶性膳食纤维的益生菌递送微胶囊及制备方法 Download PDFInfo
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Abstract
本发明公开了一种基于香菇可溶性膳食纤维的益生菌递送微胶囊及其制备方法,方法:(1)香菇粉碎后,乙醇水溶液提取,得到香菇残渣;(2)加水,蒸压处理,水提加碱性蛋白酶,反应,加α‑淀粉酶,反应,灭酶,得到酶解液;(3)浓缩,加入乙醇,静置,离心,干燥,得到香菇可溶性膳食纤维;(4)将益生菌水悬液与无菌海藻酸钠水溶液混合均匀,在搅拌下,滴到氯化钙水溶液中,搅拌硬化后,水洗,得到海藻酸钙微胶囊,浸入香菇可溶性膳食纤维水溶液中,搅拌,洗涤,干燥,得到基于香菇可溶性膳食纤维的益生菌递送微胶囊。本发明的微胶囊包埋率高,提高了益生菌的活性和稳定性,对人体肠道健康具有协同促进作用,操作简单、经济易行。
Description
技术领域
本发明涉及益生菌微胶囊化递送领域,具体涉及一种基于香菇可溶性膳食纤维的益生菌递送微胶囊及其制备方法。
背景技术
在人们保健意识不断增强的今天,人们更倾向于在日常饮食中选择含有益生菌的发酵食品和补品,以预防生理疾病并保持健康。益生菌通过口服传递并整合到肠道系统中,在肠道中能够永久定植的菌株将会与宿主粘膜或上皮细胞、免疫系统和自生微生物群密切相互作用。它们在维持人类健康,促进肠道菌群的健康调节以及参与治疗各种疾病,如糖尿病,肥胖症,非酒精性脂肪肝,高脂血症等方面,发挥着重要作用。植物乳杆菌作为代表性的乳酸菌,其商业用途历史悠久,目前被广泛用于食品、保健品或药物中。植物乳杆菌主要与降低收缩压和舒张压、抑制脂肪堆积、降低胆固醇水平、恢复胃肠道疾病、预防腹泻、减轻炎症性肠病和肠易激综合征有关。
美国食品药品监督管理局和食品工业通常建议,益生菌食品中的最小细胞数应至少为106CFU/mL,并具有较高的存活率,从而可以实现益生菌的预期健康效果。但是,大多数益生菌的口服方式会由于体内高浓度的酸和胆汁盐使益生菌活力降低,同时生产和储存过程也会造成益生菌活力的损失,从而使益生菌的功效降低。
在大规模生产中,各种益生菌微囊化技术应运而生,例如挤压、乳化、流化床、静电纺丝、撞击气溶胶、冷冻、喷雾干燥等。微囊化可以将益生菌限制在聚合物基质内,将它们与外界环境物理隔离,在恶劣条件下提供保护并能有效控制益生菌的释放。海藻酸钠-钙微胶囊由于其无毒、生物相容性好和成本低的特性而广泛用于微生态制剂的封装。然而,由于该微胶囊具有多孔结构且对酸性环境敏感,因此,海藻酸钠-钙微胶囊的缺陷需要其他材料更有益的方式弥补。
利用益生元和益生菌之间的协同作用,合生元可以通过高存活率的微生物膳食补充剂来有益地影响宿主胃肠道。可溶性膳食纤维作为益生元由于其不能被胃肠道吸收的特性引起了研究人员的广泛关注。可溶性膳食纤维的摄入可以增强饱腹感,刺激食欲抑制激素的释放,在肠道内发酵后对宿主肠道菌群有积极的影响。
香菇中富含纤维而被认为是食品、药品、保健产品和其他补品中益生元的良好替代来源。香菇可溶性膳食纤维能有效影响宿主肠道中拟杆菌、双歧杆菌等菌群的生长,但目前,尚未有一种基于香菇可溶性膳食纤维的益生菌递送微胶囊及其制备方法的报道。
发明内容
本发明的目的是克服现有技术的不足,提供能提升益生菌在口服递送过程中的存活率的一种基于香菇可溶性膳食纤维的益生菌递送微胶囊。
本发明的第二个目的是提供一种基于香菇可溶性膳食纤维的益生菌递送微胶囊的制备方法。
本发明的技术方案概述如下:
一种基于香菇可溶性膳食纤维的益生菌递送微胶囊的制备方法,包括如下步骤:
(1)香菇(Lentinus edodes)粉碎后,加入乙醇水溶液,加热回流提取,过滤,固体干燥,得到香菇残渣;
(2)将香菇残渣与去离子水混合,使含水质量为20%~30%,置于高压釜中,在115~120℃、0.1~0.2MPa下蒸压处理15~45min,得到蒸压处理后的香菇物料;加入相当于蒸压处理后的香菇物料15~25质量倍的去离子水,在80~100℃提取0.5~2h,过滤,再提取2~3次,合并滤液,调节pH为8.0~10.5,加入相当蒸压处理后的香菇物料质量的0.1%~0.5%的碱性蛋白酶,在50~60℃下水浴30~60min,再调节pH为5.5~7.0,加入相当蒸压处理后的香菇物料质量的1%~5%的α-淀粉酶,水浴30~60min,灭酶,冷却至室温后过滤,收集滤液得到酶解液;
(3)将酶解液减压浓缩至所述酶解液体积的1/10~1/4,得到浓缩液;向浓缩液中加入无水乙醇,使乙醇的终体积浓度为10%-60%,静置8~12h,在2000~4000r/min的转速下离心10~20分钟,收集沉淀物;用乙醇水溶液对沉淀物洗涤脱色,在2000~4000r/min的转速下离心10~20分钟,收集沉淀物,干燥,得到香菇可溶性膳食纤维;
(4)将浓度为1×106~1×108CFU/mL益生菌水悬液与质量浓度为1.5%~4%的无菌海藻酸钠水溶液按体积比1:1~5的比例,混合均匀,得混合液,在搅拌下,将混合液滴入到质量浓度为2%~5%的无菌氯化钙水溶液中,继续搅拌30~35min硬化后,用无菌水洗涤,得到海藻酸钙微胶囊,将海藻酸钙微胶囊浸入质量浓度为2%~5%的无菌香菇可溶性膳食纤维水溶液中,在100~200r/min下持续搅拌10~15min,用无菌生理盐水洗涤,真空冷冻干燥,得到基于香菇可溶性膳食纤维的益生菌递送微胶囊。
步骤(1)优选为:将香菇(Lentinus edodes)粉碎后,按照料液比1:15~25的比例,加入体积浓度50%~80%的乙醇水溶液,加热回流提取1~2h,过滤,残渣再重复加热回流提取2~3次,过滤,固体干燥,得到香菇残渣。
步骤(2)优选为:将香菇残渣与去离子水混合,使含水质量为30%,置于高压釜中,在120℃、0.1MPa下蒸压处理30min,得到蒸压处理后的香菇物料;加入相当于蒸压处理后的香菇物料20质量倍的去离子水,在100℃提取2h,过滤,再提取2次,合并滤液,调节pH为9.0,加入相当蒸压处理后的香菇物料质量的0.2%的碱性蛋白酶,在55℃下水浴30min,再调节pH为6.5,加入相当蒸压处理后的香菇物料质量的2.5%的α-淀粉酶,水浴30min,灭酶,冷却至室温后过滤,收集滤液得到酶解液。
步骤(3)优选为:将酶解液减压浓缩至所述酶解液体积的1/10,得到浓缩液;向浓缩液中加入无水乙醇,使乙醇的终体积浓度为10%,静置12h,在3000r/min的转速下离心20分钟,收集沉淀物;用体积浓度为75%的乙醇水溶液对沉淀物洗涤脱色,在3000r/min的转速下离心20分钟,收集沉淀物,干燥,得到香菇可溶性膳食纤维。
步骤(4)优选为:将浓度为1×108CFU/mL益生菌水悬液与质量浓度为1.5%的无菌海藻酸钠水溶液按体积比1:1的比例,混合均匀,得混合液,在搅拌下,将混合液滴入到质量浓度为2%的无菌氯化钙水溶液中,继续搅拌30min硬化后,用无菌水洗涤,得到海藻酸钙微胶囊,将海藻酸钙微胶囊浸入质量浓度为2%的无菌香菇可溶性膳食纤维水溶液中,在100r/min下持续搅拌10min,用无菌生理盐水洗涤,真空冷冻干燥,得到基于香菇可溶性膳食纤维的益生菌递送微胶囊。
益生菌为长双歧杆菌、短双歧杆菌、植物乳杆菌和鼠李糖乳杆菌中的至少一种。
上述制备方法制备的基于香菇可溶性膳食纤维的益生菌递送微胶囊。
本发明的有益效果:
(1)本发明首次采用高压釜处理香菇残渣物料,适宜条件下的高温高压处理会破坏氢键和疏水力作用而不破坏主链上的共价键,进而有效提高菇中的可溶性膳食纤维提取过程中的产量。
(2)本发明的益生菌递送微胶囊的壁材采用可溶性膳食纤维,提取自香菇残渣物料。香菇生长速度快,产量占世界可食用蘑菇的25%,来源充足。对其加以利用可以减少自然资源的大量浪费。同时作为天然来源的生物大分子,生物相容性好、无毒副作用。
(3)本发明的一种基于香菇可溶性膳食纤维的益生菌递送微胶囊包封率好,单位质量微胶囊中的益生菌数量大于106CFU,有足够数量的益生菌存活发挥其益生菌功效。
(4)本发明采用香菇可溶性膳食纤维为壁材,通过微胶囊技术对益生菌进行包埋,延长了益生菌在室温下的存储和高温环境中的活性,提高了抵抗胃肠道复杂环境的能力,提高了益生菌的活性和稳定性。
(5)本发明的一种基于香菇可溶性膳食纤维的益生菌递送微胶囊在大肠中发酵会产生有益的代谢产物,如短链脂肪酸等,同时也作为益生元在发酵过程中选择性促进健康相关有益细菌(例如双歧杆菌属和乳杆菌属)的增殖和代谢活性,还可以通过竞争营养和粘附位点来预防肠道疾病潜在的病原菌定植,有助于促进和维持肠道健康。
(6)本发明的香菇可溶性膳食纤维与被包埋的益生菌一起,对促进人体健康具有协同作用。
(7)本发明可在酸奶、牛奶、益生菌制剂等食品、保健品中应用。
附图说明
图1为本发明一种基于香菇可溶性膳食纤维的益生菌递送微胶囊(简称益生菌递送微胶囊)激光共聚焦图;
图2为本发明益生菌递送微胶囊横截面电镜图;
图3为本发明益生菌递送微胶囊热稳定性;
图4为本发明益生菌递送微胶囊吸湿性;
图5为本发明益生菌递送微胶囊在模拟胃肠道条件下的益生菌活性损失率。
具体实施方式
下面结合具体实施例对本发明作进一步的说明。
本发明使用的所有原料均为商品:
香菇:购自天津青光蔬菜基地;
碱性蛋白酶:购于天津诺澳酶制剂技术有限公司;
α-淀粉酶:购于北京索莱宝科技有限公司;
氯化钙:购于天津市江天化工技术有限公司;
海藻酸钠:购自天津市江天化工技术有限公司;
无水乙醇:购自天津市江天化工技术有限公司;
MRS肉汤(MRS液体培养基粉末):购自北京索莱宝生物科技有限公司;
植物乳杆菌LP90:购自江苏微康生物科技有限公司。
上述原料的介绍是为了使本领域的技术人员能够更好地理解本发明,但并不对本发明作任何限制,其它企业生产的上述原料,也可以用于本发明。
实施例1
一种基于香菇可溶性膳食纤维的益生菌递送微胶囊的制备方法,包括如下步骤:
(1)为将香菇(Lentinus edodes)粉碎后,按照料液比1:20的比例,加入体积浓度75%的乙醇水溶液,加热回流提取1.5h,过滤,残渣再重复加热回流提取3次,过滤,固体干燥,得到香菇残渣;
(2)将香菇残渣与去离子水混合,使含水质量为30%,置于高压釜中,在120℃、0.1MPa下蒸压处理30min,得到蒸压处理后的香菇物料;加入相当于蒸压处理后的香菇物料20质量倍的去离子水,在100℃提取2h,过滤,再提取2次,合并滤液,调节pH为9.0,加入相当蒸压处理后的香菇物料质量的0.2%的碱性蛋白酶,在55℃下水浴30min,再调节pH为6.5,加入相当蒸压处理后的香菇物料质量的2.5%的α-淀粉酶,水浴30min,100℃灭酶5min,冷却至室温后过滤,收集滤液得到酶解液;
(3)将酶解液减压浓缩至所述酶解液体积的1/10,得到浓缩液;向浓缩液中加入无水乙醇,使乙醇的终体积浓度为10%,静置12h,在3000r/min的转速下离心20分钟,收集沉淀物;用体积浓度为75%的乙醇水溶液对沉淀物洗涤脱色,在3000r/min的转速下离心20分钟,收集沉淀物,干燥,得到香菇可溶性膳食纤维;
(4)将浓度为1×108CFU/mL益生菌(植物乳杆菌LP90)水悬液与质量浓度为1.5%的无菌海藻酸钠水溶液按体积比1:1的比例,混合均匀,得混合液,在搅拌下,将混合液滴入到质量浓度为2%的无菌氯化钙水溶液中,继续搅拌30min硬化后,用无菌水洗涤,得到海藻酸钙微胶囊,将海藻酸钙微胶囊浸入质量浓度为2%的无菌香菇可溶性膳食纤维水溶液中,在100r/min下持续搅拌10min,用无菌生理盐水洗涤,真空冷冻干燥,得到基于香菇可溶性膳食纤维的益生菌递送微胶囊,简称C-LES-1,益生菌包封率为95.67%。
本实施例的益生菌水悬液为植物乳杆菌LP90水悬液,制备步骤如下:
将保存于-20℃条件下的植物乳杆菌菌粉接种于121±1℃灭菌15min冷却后的MRS液体培养基中,在36±1℃条件下,震荡培养24h,进行初次活化;(MRS液体培养基:市售的MRS肉汤按其说明书配制;)
按质量浓度为5%的比例将初次活化的植物乳杆菌培养液再次接入MRS液体培养基中活化,在36±1℃条件下,震荡培养24h(达到植物乳杆菌生长对数期),得到植物乳杆菌培养物;
将植物乳杆菌培养物在3000r/min的转速下离心2min,除去上清液,用无菌生理盐水洗涤2次,用无菌水调节植物乳杆菌浓度至1×108CFU/mL,得到益生菌(植物乳杆菌)水悬液。
实施例2
一种基于香菇可溶性膳食纤维的益生菌递送微胶囊的制备方法,包括如下步骤:
(1)为将香菇(Lentinus edodes)粉碎后,按照料液比1:15的比例,加入体积浓度80%的乙醇水溶液,加热回流提取2h,过滤,残渣再重复加热回流提取2次,过滤,固体干燥,得到香菇残渣;
(2)将香菇残渣与去离子水混合,使含水质量为20%,置于高压釜中,在115℃、0.1MPa下蒸压处理45min,得到蒸压处理后的香菇物料;加入相当于蒸压处理后的香菇物料15质量倍的去离子水,在80℃提取2h,过滤,再提取3次,合并滤液,调节pH为10.5,加入相当蒸压处理后的香菇物料质量的0.1%的碱性蛋白酶,在50℃下水浴60min,再调节pH为5.5,加入相当蒸压处理后的香菇物料质量的1%的α-淀粉酶,水浴60min,100℃灭酶5min,冷却至室温后过滤,收集滤液得到酶解液;
(3)将酶解液减压浓缩至所述酶解液体积的1/10,得到浓缩液;向浓缩液中加入无水乙醇,使乙醇的终体积浓度为30%,静置12h,在2000r/min的转速下离心20分钟,收集沉淀物;用体积浓度为75%的乙醇水溶液对沉淀物洗涤脱色,在2000r/min的转速下离心20分钟,收集沉淀物,干燥,得到香菇可溶性膳食纤维;
(4)将浓度为1×106CFU/mL益生菌(植物乳杆菌LP90)水悬液与质量浓度为1.5%的无菌海藻酸钠水溶液按体积比1:1的比例,混合均匀,得混合液,在搅拌下,将混合液滴入到质量浓度为2%的无菌氯化钙水溶液中,继续搅拌35min硬化后,用无菌水洗涤,得到海藻酸钙微胶囊,将海藻酸钙微胶囊浸入质量浓度为2%的无菌香菇可溶性膳食纤维水溶液中,在100r/min下持续搅拌10min,用无菌生理盐水洗涤,真空冷冻干燥,得到基于香菇可溶性膳食纤维的益生菌递送微胶囊,简称C-LES-2,益生菌包封率为91.67%。
本实施例的益生菌水悬液为植物乳杆菌LP90水悬液,制备步骤与实施例1相同,用无菌水调节植物乳杆菌浓度至1×106CFU/mL,得到益生菌(植物乳杆菌)水悬液。
实施例3
一种基于香菇可溶性膳食纤维的益生菌递送微胶囊的制备方法,包括如下步骤:
(1)为将香菇(Lentinus edodes)粉碎后,按照料液比1:25的比例,加入体积浓度50%的乙醇水溶液,加热回流提取1h,过滤,残渣再重复加热回流提取3次,过滤,固体干燥,得到香菇残渣;
(2)将香菇残渣与去离子水混合,使含水质量为25%,置于高压釜中,在118℃、0.2MPa下蒸压处理15min,得到蒸压处理后的香菇物料;加入相当于蒸压处理后的香菇物料25质量倍的去离子水,在100℃提取0.5h,过滤,再提取2次,合并滤液,调节pH为8.5,加入相当蒸压处理后的香菇物料质量的0.5%的碱性蛋白酶,在60℃下水浴30min,再调节pH为7.0,加入相当蒸压处理后的香菇物料质量的5%的α-淀粉酶,水浴30min,100℃灭酶5min,冷却至室温后过滤,收集滤液得到酶解液;
(3)将酶解液减压浓缩至所述酶解液体积的1/4,得到浓缩液;向浓缩液中加入无水乙醇,使乙醇的终体积浓度为60%,静置8h,4000r/min的转速下离心10分钟,收集沉淀物;用体积浓度为75%的乙醇水溶液对沉淀物洗涤脱色,在4000r/min的转速下离心10分钟,收集沉淀物,干燥,得到香菇可溶性膳食纤维;
(4)将浓度为1×108CFU/mL益生菌(植物乳杆菌LP90)水悬液与质量浓度为4%的无菌海藻酸钠水溶液按体积比1:5的比例,混合均匀,得混合液,在搅拌下,将混合液滴入到质量浓度为5%的无菌氯化钙水溶液中,继续搅拌30min硬化后,用无菌水洗涤,得到海藻酸钙微胶囊,将海藻酸钙微胶囊浸入质量浓度为5%的无菌香菇可溶性膳食纤维水溶液中,在200r/min下持续搅拌15min,用无菌生理盐水洗涤,真空冷冻干燥,得到基于香菇可溶性膳食纤维的益生菌递送微胶囊,简称C-LES-3,益生菌包封率为92.22%。
本实施例的益生菌水悬液的制备同实施例1的益生菌水悬液的制备。
实验证明,用长双歧杆菌、短双歧杆菌、鼠李糖乳杆菌或长双歧杆菌和短双歧杆菌的混合菌替代本实施例的植物乳杆菌LP90,其它同本实施例,可以分别制备出相应的基于香菇可溶性膳食纤维的益生菌递送微胶囊。
对照例
未使用香菇可溶性膳食纤维作为壁材的益生菌递送微胶囊制备:
将浓度为1×108CFU/mL的植物乳杆菌LP90水悬液与质量浓度为1.5%的无菌的海藻酸钠水溶液按体积比1:1的比例,混合均匀,得混合液,在搅拌下,将混合液滴入到质量浓度为2%的无菌的氯化钙水溶液中,继续搅拌30min硬化后,用无菌水洗涤,真空冷冻干燥,得到未使用香菇可溶性膳食纤维作为壁材的益生菌递送微胶囊,简称Non-SDF,益生菌包封率为93.09%。
本对照例的益生菌水悬液的制备同实施例1益生菌水悬液的制备。
实施例4
基于香菇可溶性膳食纤维的益生菌递送微胶囊的热稳定性测定。
实验方法:
(1)将植物乳杆菌LP90以及各实施例和对照例制备的益生菌递送微胶囊各0.1g,分别与提前预热至45℃、55℃和65℃的9mL无菌水混合,反应30分钟,冷却至室温。
(2)取步骤(1)获得的0.1g各实施例、对照例的益生菌递送微胶囊与9.9mL柠檬酸钠二水合物水溶液(质量浓度10%,pH=8.2)混合直至其完全解离;将解离后得到的益生菌用无菌去离子水稀释至浓度为30~300CFU/mL范围内得到菌悬液,菌悬液滴于MRS固体培养基(平板)上,涂匀,待琼脂凝固后,将平板翻转,36±1℃厌氧培养72±2h。培养后计数平板上的所有菌落数。换算出微胶囊单位体积的活菌数,其结果见图3。
将取步骤(1)获得的0.1g植物乳杆菌LP90用无菌去离子水稀释至浓度为30~300CFU/mL范围内得到菌悬液,菌悬液滴于MRS固体培养基(平板)上,涂匀,待琼脂凝固后,将平板翻转,36±1℃厌氧培养72±2h。培养后计数平板上的所有菌落数。换算出微胶囊单位体积的活菌数,其结果见图3。
活菌数计算公式:活菌数(CFU/mL)=同一稀释度3个重复MRS固体培养基菌落平均数×稀释倍数。
由上述实验结果可以看出,与对照例相比,基于香菇可溶性膳食纤维的益生菌递送微胶囊在45℃、55℃、65℃的高温下细胞活力显著提高。表明本发明制备的益生菌递送微胶囊具有显着的耐热性优势,并且可以在益生菌细胞的热处理过程中提供更好的保护。
实施例5
一种基于香菇可溶性膳食纤维的益生菌递送微胶囊的吸湿性测定。
实验方法:将各实施例和对照例制备的益生菌递送微胶囊暴露于80%相对湿度、温度为22℃至25℃的密闭容器中。每隔3、6、9h测量一次水分增加。微胶囊的吸湿性表示为干重的百分比,其结果见图4。
由上述实验结果可以看出,在80%湿度环境下存储9h后,基于香菇可溶性膳食纤维的益生菌递送微胶囊的水分含量显著低于对照例,吸湿性增幅较低。
实施例6
基于香菇可溶性膳食纤维的益生菌递送微胶囊在模拟胃肠道条件下的存活率测定。
实验方法:将1g各实施例和对照例制备的益生菌递送微胶囊分别与9mL人工胃液混合,在36±1℃下孵育2h,在30min(G-30)、60min(G-60)和120min(G-120)的时间间隔下对活细胞计数。随后,1g经过人工胃液消化后的益生菌递送微胶囊与9mL人工肠液混合,在36±1℃下孵育2h,在60min(I-60)和120min(I-120)的时间间隔下对活细胞计数,其结果见图5。
人工胃液的制备方法为:将NaCl、KCl、CaCl2·2H2O、NaHCO3、胃蛋白酶按照31:11:2:6:3的质量比混合溶于水中,搅拌直至完全溶解,调节pH为2.5,通过0.22μm的过滤器灭菌。
人工肠液的制备方法为:将KCl、CaCl2·2H2O、NaCl、牛黄胆酸钠、胰脂肪酶按照6:3:54:4:7的质量比混合溶于水中,搅拌直至完全溶解,调节pH为6.5,通过0.22μm的过滤器灭菌。
活菌计数方法为:将0.1g各实施例、对照例制备的益生菌递送微胶囊与9.9mL柠檬酸钠二水合物水溶液(质量浓度10%,pH=8.2)混合直至其完全解离。将解离后得到的益生菌用无菌去离子水稀释至浓度为30~300CFU/mL范围内,菌悬液滴于平板上,涂匀,待琼脂凝固后,将平板翻转,36±1℃厌氧培养72±2h。培养后计数平板上的所有菌落数。换算出各实施例、对照例的微胶囊单位体积的活菌数。
活菌数计算公式:活菌数(CFU/mL)=同一稀释度3个重复固体培养基菌落平均数×稀释倍数。
由上述实验结果可以看出,与对照例相比,基于香菇可溶性膳食纤维的益生菌递送微胶囊在通过胃肠道的复杂环境后,细胞存活率显著提升。表明本发明制备的微胶囊对人造胃肠液的复杂环境具有较高的抵抗力,可提高益生菌口服递送过程的生存能力。
以上对本发明做了示例性的描述,应该说明的是,在不脱离本发明的核心的情况下,任何简单的变形、修改或者其他本领域技术人员能够不花费创造性劳动的等同替换均落入本发明的保护范围。
Claims (7)
1.一种基于香菇可溶性膳食纤维的益生菌递送微胶囊的制备方法,其特征是包括如下步骤:
(1)香菇(Lentinus edodes)粉碎后,加入乙醇水溶液,加热回流提取,过滤,固体干燥,得到香菇残渣;
(2)将香菇残渣与去离子水混合,使含水质量为20%~30%,置于高压釜中,在115~120℃、0.1~0.2MPa下蒸压处理15~45min,得到蒸压处理后的香菇物料;加入相当于蒸压处理后的香菇物料15~25质量倍的去离子水,在80~100℃提取0.5~2h,过滤,再提取2~3次,合并滤液,调节pH为8.5~10.5,加入相当蒸压处理后的香菇物料质量的0.1%~0.5%的碱性蛋白酶,在50~60℃下水浴30~60min,再调节pH为5.5~7.0,加入相当蒸压处理后的香菇物料质量的1%~5%的α-淀粉酶,水浴30~60min,灭酶,冷却至室温后过滤,收集滤液得到酶解液;
(3)将酶解液减压浓缩至所述酶解液体积的1/10~1/4,得到浓缩液;向浓缩液中加入无水乙醇,使乙醇的终体积浓度为10%-60%,静置8~12h,在2000~4000r/min的转速下离心10~20分钟,收集沉淀物;用乙醇水溶液对沉淀物洗涤脱色,在2000~4000r/min的转速下离心10~20分钟,收集沉淀物,干燥,得到香菇可溶性膳食纤维;
(4)将浓度为1×106~1×108CFU/mL益生菌水悬液与质量浓度为1.5%~4%的无菌海藻酸钠水溶液按体积比1:1~5的比例,混合均匀,得混合液,在搅拌下,将混合液滴入到质量浓度为2%~5%的无菌氯化钙水溶液中,继续搅拌30~35min硬化后,用无菌水洗涤,得到海藻酸钙微胶囊,将海藻酸钙微胶囊浸入质量浓度为2%~5%的无菌香菇可溶性膳食纤维水溶液中,在100~200r/min下持续搅拌10~15min,用无菌生理盐水洗涤,真空冷冻干燥,得到基于香菇可溶性膳食纤维的益生菌递送微胶囊。
2.根据权利要求1所述的制备方法,其特征在于所述步骤(1)为:将香菇(Lentinusedodes)粉碎后,按照料液比1:15~25的比例,加入体积浓度50%~80%的乙醇水溶液,加热回流提取1~2h,过滤,残渣再重复加热回流提取2~3次,过滤,固体干燥,得到香菇残渣。
3.根据权利要求1所述的制备方法,其特征在于所述步骤(2)为:将香菇残渣与去离子水混合,使含水质量为30%,置于高压釜中,在120℃、0.1MPa下蒸压处理30min,得到蒸压处理后的香菇物料;加入相当于蒸压处理后的香菇物料20质量倍的去离子水,在100℃提取2h,过滤,再提取2次,合并滤液,调节pH为9.0,加入相当蒸压处理后的香菇物料质量的0.2%的碱性蛋白酶,在55℃下水浴30min,再调节pH为6.5,加入相当蒸压处理后的香菇物料质量的2.5%的α-淀粉酶,水浴30min,灭酶,冷却至室温后过滤,收集滤液得到酶解液。
4.根据权利要求1所述的制备方法,其特征在于所述步骤(3)为:将酶解液减压浓缩至所述酶解液体积的1/10,得到浓缩液;向浓缩液中加入无水乙醇,使乙醇的终体积浓度为10%,静置12h,在3000r/min的转速下离心20分钟,收集沉淀物;用体积浓度为75%的乙醇水溶液对沉淀物洗涤脱色,在3000r/min的转速下离心20分钟,收集沉淀物,干燥,得到香菇可溶性膳食纤维。
5.根据权利要求1所述的制备方法,其特征在于所述步骤(4)为:将浓度为1×108CFU/mL益生菌水悬液与质量浓度为1.5%的无菌海藻酸钠水溶液按体积比1:1的比例,混合均匀,得混合液,在搅拌下,将混合液滴入到质量浓度为2%的无菌氯化钙水溶液中,继续搅拌30min硬化后,用无菌水洗涤,得到海藻酸钙微胶囊,将海藻酸钙微胶囊浸入质量浓度为2%的无菌香菇可溶性膳食纤维水溶液中,在100r/min下持续搅拌10min,用无菌生理盐水洗涤,真空冷冻干燥,得到基于香菇可溶性膳食纤维的益生菌递送微胶囊。
6.根据权利要求1或5所述的制备方法,其特征在于所述益生菌为长双歧杆菌、短双歧杆菌、植物乳杆菌和鼠李糖乳杆菌中的至少一种。
7.权利要求1-6之一制备方法制备的基于香菇可溶性膳食纤维的益生菌递送微胶囊。
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