CN115399481B - 一种载有益生菌的功能化可食性膜及其制备方法 - Google Patents
一种载有益生菌的功能化可食性膜及其制备方法 Download PDFInfo
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- CN115399481B CN115399481B CN202210859329.8A CN202210859329A CN115399481B CN 115399481 B CN115399481 B CN 115399481B CN 202210859329 A CN202210859329 A CN 202210859329A CN 115399481 B CN115399481 B CN 115399481B
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- liposome
- protamine
- probiotics
- probiotic
- edible film
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- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
本发明公开了一种载有益生菌的功能化可食性膜及其制备方法,所述的可食用膜以生物聚合物为基质,填充有鱼精蛋白聚集体保护的益生菌脂质体。活性益生菌被脂质体包裹,赋予其缓释放功能。脂质体表面经过β‑胆固醇‑D‑葡萄糖苷(β‑CG)修饰,可以协同两性离子脂质诱导鱼精蛋白通过氢键作用在脂质体周围生成球形纤维聚集体,所述鱼精蛋白纤维聚集体的直径为10‑30um。鱼精蛋白本身有较高的热稳定性,此蛋白形成的纤维聚集体在脂质体周围形成了稳定的载体保护作用,提高了益生菌活性物的热稳定性。可以克服由于益生菌热不稳定特性,仅能用湿法成膜方法生产食用膜的缺陷。
Description
技术领域
本发明涉及食品加工领域,具体涉及一种载有益生菌的功能化可食性膜及其制备方法。
背景技术
可食性膜是指由食用生物聚合物和食品添加剂通过一定的加工工艺使各成膜剂分子之间相互作用而成的具有特定功能的可食用薄膜。制成的薄膜可作为包装或覆盖材料直接应用于食品,以延长食品的保质期。随着人们对食品品质要求的提高以及环保意识的增强,功能性可食膜在人们的日常生活中将得到越来越广泛的应用。例如可在食品包装膜中添加多种活性化合物,如抗氧化剂、抗菌剂、微量营养素、天然色素、风味剂、生物活性肽、细菌细胞外次级代谢产物、合生元、益生菌和后生元等。这不仅可以帮助活性成分发挥功效,又能起到延长食品保质期的作用。
其中益生菌是一类能维护宿主机体微生态平衡、有益于机体健康的微生物。目前应用于人类的益生菌种类主要有乳杆菌、双歧杆菌、肠球菌、乳球菌和嗜热链球菌等。但是益生菌在生产、储存以及通过胃肠道的过程都会容易受到pH、氧气浓度和温度变化等的影响,使其活力下降。特别是含益生菌的可食用膜由于益生菌细胞的热不稳定特性,仅能用低温湿法成膜方法生产。因此如何保持益生菌的高活性和稳定性显得非常重要。
发明内容
针对益生菌的易失活等不稳定性问题,本发明提供一种载有益生菌的功能化可食性膜及其制备方法,其可提高活性成分的稳定性、递送和吸收效率以及应用价值。
本发明还提供一种鱼精蛋白聚集体保护的益生菌脂质体,是一种稳定性能好、胞内递送效率高、具有一定抗菌能力的纳米粒子。
具体技术方案如下:
一种载有益生菌的功能化可食性膜,以质量百分比计,原料组成包括:醇溶蛋白25-30%、海藻多糖15-20%、增塑剂1-3%、鱼精蛋白聚集体保护的益生菌脂质体0.1-0.5%,余量为溶剂。
醇溶蛋白包括玉米醇溶蛋白、小麦醇溶蛋白等中的至少一种;
海藻多糖采用琼脂、海藻酸钠和壳聚糖中的其中一种或者几种混合;
所述增塑剂采用甘油、丙二醇、山梨醇、木糖醇、甘露醇、乙二醇、葡萄糖和果糖等中的至少一种;
所述鱼精蛋白聚集体保护的益生菌脂质体是鱼精蛋白在脂质体表面形成球形聚集体保护益生菌脂质体。益生菌加入到由类脂、β-胆固醇-D-葡萄糖苷和抗氧化剂形成的脂质薄膜中,得到益生菌脂质体,鱼精蛋白在脂质体表面形成球形聚集体保护益生菌脂质体。
采用β-胆固醇-D-葡萄糖苷(β-CG)修饰,协同两性离子脂质诱导鱼精蛋白纤维在脂质体表面形成球形聚集体,脂质体和鱼精蛋白之间主要通过氢键网络连接。
所述的载有益生菌的功能化可食性膜通过流延法制备,按如下步骤进行:
S1,将醇溶蛋白加入到乙醇溶液中,在热水浴中搅拌至溶解;
S2,将海藻多糖加入到冰醋酸中,在热水浴中搅拌至溶解;
S3,将上述溶液混合均匀后,加入增塑剂和鱼精蛋白聚集体保护的益生菌脂质体,充分搅拌,于热水浴中交联反应一段时间,真空脱气消泡;
S4,将溶液浇注于洁净干燥的聚四氟乙烯成膜版上,进行热处理固膜,制得载有益生菌的功能化可食性膜。
所述S1中,醇溶蛋白-乙醇溶液质量浓度为9%-13%;热水浴温度为40-50℃。
所述S2中,海藻多糖-醋酸溶液质量浓度为3%-5%;热水浴温度为40-50℃。
所述S3中,交联反应条件为50℃-55℃,水浴中反应1-2h;
所述S4中,热处理条件为90-130℃,处理时间为10-20min。
本发明还提供一种鱼精蛋白聚集体保护的益生菌脂质体,益生菌加入到由类脂、β-胆固醇-D-葡萄糖苷和抗氧化剂形成的脂质薄膜中,得到益生菌脂质体,鱼精蛋白在脂质体表面形成球形聚集体保护益生菌脂质体。
其中:类脂为两性离子脂质,包括大豆卵磷脂、蛋黄卵磷脂、1-棕榈酰-2-油酰-磷脂酰胆碱(POPC)、二肉豆蔻酰磷脂酰胆碱(DMPC)中的一种或几种。所述抗氧化剂为VE乙酸酯、没食子酸丙酯中的任一种。类脂和β-CG的摩尔比3.5-4:1,类脂和益生菌的质量比为6-15:1,类脂和鱼精蛋白的摩尔比1.3-2:1。
鱼精蛋白聚集体保护的益生菌脂质体粒径10-30um,分布均匀。
所述鱼精蛋白聚集体保护的益生菌脂质体的制备,包括如下步骤:
(1)称取类脂、β-胆固醇-D-葡萄糖苷(β-CG)和抗氧化剂在有机溶剂中溶解,溶剂在旋转蒸发器中蒸发除去,形成脂质薄膜,随后在真空下干燥过夜;
(2)将益生菌培养物悬浮于磷酸盐缓冲液中,得到细菌溶液;
(3)将步骤(2)中的溶液加入到(1)中脂质薄膜中,水合并涡旋形成脂质体混悬液,经高压均质后得到益生菌脂质体悬浮液;
(4)将鱼精蛋白粉末溶于去离子水中,之后与脂质体悬浮液混合均匀,搅拌孵育一段时间,制备得鱼精蛋白聚集体保护的益生菌脂质体乳液。
所述(1)的类脂为两性离子脂质,包括大豆卵磷脂、蛋黄卵磷脂、1-棕榈酰-2-油酰-磷脂酰胆碱(POPC)、二肉豆蔻酰磷脂酰胆碱(DMPC)中的一种或几种。所述溶剂优选包括氯仿、甲醇、乙醇中的至少一种;所述抗氧化剂为VE乙酸酯、没食子酸丙酯中的任一种;抗氧化剂添加量为脂质体质量的0.5%-1%;所述类脂和β-CG的摩尔比3.5-4:1。
优选地,步骤(1)中,减压旋蒸的温度为40~50℃,减压旋蒸的时间为30~60min,减压旋蒸的转速为50~150rpm。
优选地,步骤(2)中所述益生菌在磷酸盐缓冲溶液的浓度为0.01~0.03mg/mL;益生菌选自乳杆菌属、双歧杆菌属、肠球菌属、大肠杆菌属、芽孢杆菌属、丁酸梭菌属和酵母菌属中的一种或多种菌株;
所述磷酸盐缓冲液pH为7.5。
所述步骤(3)中,类脂和益生菌的质量比为6-15:1;所述高压均质处理的参数条件是:300~800bar压力下,均质1~10次。
所述步骤(4)中搅拌孵育的条件为室温下孵育16-24h。
所述步骤(4)中的鱼精蛋白水溶液浓度为0.2~0.5mg/ml,类脂和鱼精蛋白的摩尔比1.3-2:1。
制备的在益生菌脂质体周围生成的鱼精蛋白聚集体粒径分布均匀,只有10-30um,远小于其他蛋白聚集体微胶囊(1-10mm)。
本发明中,鱼精蛋白是一种碱性蛋白质,有很高的营养性和功能性,能降血压、促消化等,具有很强的抑菌能力,并有较高的热稳定性,在210℃条件下加热1小时仍具有活性,借鉴于鱼精蛋白耐热稳定性能,本发明制备了益生菌脂质体,并经鱼精蛋白纤维聚集体保护后,其耐热性能和存活率大大提高,可满足各种成膜加工方法的要求。
有益效果
本发明将益生菌脂质体整合在可食膜中,并通过对益生菌脂质体耐热性能改善后,将脂质体的制备工艺与可食用膜的制备工艺结合,完全可满足各种成膜加工方法的要求。制备的可食膜抑菌和防腐保鲜效果好,可以方便夹在汉堡、饼干、糕点中等,因此极大方便了益生菌与其它食品的配合食用。
本发明提供一种脂质体,经过β-胆固醇-D-葡萄糖苷(β-CG)修饰后,可以协同两性离子脂质诱导鱼精蛋白球形纤维聚集体的形成,此时脂质体和鱼精蛋白之间主要通过氢键网络的作用。使用该方式封装益生菌后活菌存活率和热稳定性得到显著提高。
附图说明
图1为鱼精蛋白在益生菌脂质体表面形成球形纤维聚集体的机制示意图;
图2为鱼精蛋白聚集体保护的益生菌脂质体的全内反射荧光显微镜图;
图3为实施例1的益生菌脂质体粒径分布曲线图。
图4为活性菌数对比。
图5为益生菌存活率对比。
具体实施方式
实施例1
以质量百分比计,本实施例的载有双歧杆菌的功能化可食性膜原料组成包括:玉米醇溶蛋白25%、壳聚糖15%、甘油1%、鱼精蛋白聚集体保护的双歧杆菌脂质体0.1%,余量为溶剂。
所述的载有双歧杆菌的功能化可食性膜通过流延法制备,按如下步骤进行:
步骤1,将10g玉米醇溶蛋白加入到90g乙醇溶液中,在50℃热水浴中搅拌至溶解,配制质量浓度为10%的玉米醇溶蛋白-乙醇溶液;
步骤2,将6g壳聚糖加入到114g冰醋酸中,在50℃热水浴中搅拌至溶解,配制质量浓度为5%的玉米醇溶蛋白-乙醇溶液;
步骤3,取上述玉米醇溶蛋白-乙醇溶液和壳聚糖-冰醋酸溶液混合均匀后,加入0.4g甘油和0.04g鱼精蛋白聚集体保护的双歧杆菌脂质体,300r/min充分搅拌,于55℃热水浴中交联反应1h,真空脱气消泡;
步骤4,将溶液浇注于洁净干燥的聚四氟乙烯成膜版上,进行120℃热处理10min固膜,制得载有双歧杆菌的功能化可食性膜。
所述鱼精蛋白聚集体保护的双歧杆菌脂质体,合成方法包括如下步骤:
(1)称取15.16mg大豆卵磷脂、13.56mgDMPC、3.87mg的β-CG和0.36mg的VE乙酸酯在氯仿中溶解,之后在40℃水浴旋转蒸发器中以150rpm的转速蒸发30min除去溶剂,形成脂质薄膜,随后在真空下干燥过夜;
(2)将3.59mg双歧杆菌培养物悬浮于140mL的pH为7.5的磷酸盐缓冲液中,得到浓度为0.026mg/mL的双歧杆菌溶液;
(3)将步骤(2)中的溶液加入到(1)中脂质薄膜中,水合并涡旋形成脂质体混悬液,经800bar高压均质5遍后得到双歧杆菌脂质体悬浮液;
(4)将6.62mg鱼精蛋白粉末溶于20mL去离子水中制备浓度为0.33mg/mL的混合液,之后与脂质体悬浮液混合均匀,搅拌孵育20h,制备得浓度为0.27mg/mL鱼精蛋白聚集体保护的双歧杆菌脂质体乳液。
实施例2
以质量百分比计,本实施例的载有大肠杆菌的功能化可食性膜原料组成包括:玉米醇溶蛋白28%、壳聚糖17%、甘露醇2%、鱼精蛋白聚集体保护的大肠杆菌脂质体0.4%,余量为溶剂。
所述的载有大肠杆菌的功能化可食性膜通过流延法制备,按如下步骤进行:
步骤1,将2.1g玉米醇溶蛋白加入到14g乙醇溶液中,在45℃热水浴中搅拌至溶解,配制质量浓度为13%的玉米醇溶蛋白-乙醇溶液;
步骤2,将1.27g壳聚糖加入到24.13g冰醋酸中,在50℃热水浴中搅拌至溶解,配制质量浓度为5%的壳聚糖-冰醋酸溶液;
步骤3,将上述溶液混合均匀后,加入0.15g甘露醇和0.03g鱼精蛋白聚集体保护的大肠杆菌脂质体,300r/min充分搅拌,于50℃热水浴中交联反应1.5h,真空脱气消泡;
步骤4,将溶液浇注于洁净干燥的聚四氟乙烯成膜版上,进行100℃热处理20min固膜,制得载有双歧杆菌的功能化可食性膜。
所述的脂质体为鱼精蛋白聚集体保护的大肠杆菌脂质体,合成方法包括如下步骤:
(1)称取4.71mg蛋黄卵磷脂、19mg的POPC、3.87mg的β-CG和0.15mg的VE乙酸酯在氯仿中溶解,溶剂在50℃水浴旋转蒸发器中转速100rpm蒸发40min除去,形成脂质薄膜,随后在真空下干燥过夜;
(2)将2.37mg大肠杆菌培养物悬浮于100mL的pH为7.5的磷酸盐缓冲液中,得到浓度为0.024mg/mL的双歧杆菌溶液;
(3)将步骤(2)中的溶液加入到(1)中脂质薄膜中,水合并涡旋形成脂质体混悬液,经800bar高压均质5遍后得到大肠杆菌脂质体悬浮液;
(4)将8.28mg鱼精蛋白粉末溶于20mL去离子水中制备浓度为0.41mg/mL的混合液,之后与脂质体悬浮液混合均匀,搅拌孵育24h,制备得浓度为0.32mg/mL鱼精蛋白聚集体保护的大肠杆菌脂质体乳液。
对照例1
与实施例1的区别在于:双歧杆菌脂质体未经添加β-CG。
双歧杆菌脂质体具体合成方法包括如下步骤:
(1)称取15.16mg大豆卵磷脂、13.56mgDMPC和0.36mg的VE乙酸酯在氯仿中溶解,之后在40℃水浴旋转蒸发器中以150rpm的转速蒸发30min除去溶剂,形成脂质薄膜,随后在真空下干燥过夜。
(2)将3.59mg双歧杆菌培养物悬浮于140mL的PH为7.5的磷酸盐缓冲液中,得到浓度为0.026mg/mL的双歧杆菌溶液;
(3)将步骤(2)中的溶液加入到(1)中脂质薄膜中,水合并涡旋形成脂质体混悬液,经800bar高压均质5遍后得到双歧杆菌脂质体悬浮液;
(4)将6.62mg鱼精蛋白粉末溶于20mL去离子水中制备浓度为0.33mg/mL的混合液,之后与脂质体悬浮液混合均匀,搅拌孵育20h,制备得浓度为0.27mg/mL的双歧杆菌脂质体乳液。
对照例2
与实施例1的区别在于:双歧杆菌脂质体未经鱼精蛋白聚集体保护。
双歧杆菌脂质体具体合成方法包括如下步骤:
(1)称取15.16mg大豆卵磷脂、13.56mgDMPC、3.87mg的β-CG和0.36mg的VE乙酸酯在氯仿中溶解,之后在40℃水浴旋转蒸发器中以150rpm的转速蒸发30min除去溶剂,形成脂质薄膜,随后在真空下干燥过夜。
(2)将3.59mg双歧杆菌培养物悬浮于140mL的PH为7.5的磷酸盐缓冲液中,得到浓度为0.026mg/mL的双歧杆菌溶液;
(3)将步骤(2)中的溶液加入到(1)中脂质薄膜中,水合并涡旋形成脂质体混悬液,经800bar高压均质5遍。
其余均相同。制备得到双歧杆菌脂质体乳液。
对照例3
与实施例1的区别在于:双歧杆菌脂质体未经鱼精蛋白聚集体保护,且固膜热处理温度为50℃。
以质量百分比计,本实施例的载有双歧杆菌的功能化可食性膜原料组成包括:玉米醇溶蛋白25%、壳聚糖15%、甘油1%、双歧杆菌脂质体0.1%,余量为溶剂。
所述的载有双歧杆菌的功能化可食性膜通过流延法制备,按如下步骤进行:
步骤1,将10g玉米醇溶蛋白加入到90g乙醇溶液中,在50℃热水浴中搅拌至溶解,配制质量浓度为10%的玉米醇溶蛋白-乙醇溶液;
步骤2,将6g壳聚糖加入到114g冰醋酸中,在50℃热水浴中搅拌至溶解,配制质量浓度为5%的玉米醇溶蛋白-乙醇溶液;
步骤3,取上述玉米醇溶蛋白-乙醇溶液和壳聚糖-冰醋酸溶液混合均匀后,加入0.4g甘油和0.04g鱼精蛋白聚集体保护的双歧杆菌脂质体,300r/min充分搅拌,于55℃热水浴中交联反应1h,真空脱气消泡;
步骤4,将溶液浇注于洁净干燥的聚四氟乙烯成膜版上,进行50℃热处理10min固膜,制得载有双歧杆菌的功能化可食性膜。
本发明提供所述的脂质体为双歧杆菌脂质体,合成方法包括如下步骤:
(1)称取15.16mg大豆卵磷脂、13.56mgDMPC、3.87mg的β-CG和0.36mg的VE乙酸酯在氯仿中溶解,之后在40℃水浴旋转蒸发器中以150rpm的转速蒸发30min除去溶剂,形成脂质薄膜,随后在真空下干燥过夜。
(2)将3.59mg双歧杆菌培养物悬浮于140mL的pH为7.5的磷酸盐缓冲液中,得到浓度为0.026mg/mL的双歧杆菌溶液;
(3)将步骤(2)中的溶液加入到(1)中脂质薄膜中,水合并涡旋形成脂质体混悬液,经800bar高压均质5遍后得到双歧杆菌脂质体体乳液。
测试过程
1、全内反射荧光显微镜测试(TIRFM)
为观察鱼精蛋白纤维在脂质体表面形成球形聚集体的行为,将实施例1制备的样品乳液与等量的浓度为5uM的硫黄素T磷酸缓冲液混合均匀,硫黄素T可以特异性地与纤维聚集体结合,吸取14uL混合样用全内反射荧光显微镜以观察聚集体的荧光图像
结果如图2所示,可以观察到线状纤维经硫黄素T染色后,显现荧光特征,鱼精蛋白纤维聚集在脂质体的周围,形成球形颗粒状,大小为20um左右。
2、粒径和zeta电位测试
取实施例1、实施例2和对照例1的益生菌脂质体乳液用去离子水稀释至10倍,用纳米粒度仪及zeta电位分析仪测定。
测得的益生菌脂质体的平均粒径和zeta电位见下表1,实施例和对照例制备的益生菌脂质体平均粒径皆在100-130nm左右,实施例1的益生菌脂质体粒径分布曲线图见图3所示,而实施例的zeta电位绝对值皆大于20mV,明显比对照例1的绝对值要高。这说明,β-CG的加入可以适当提高脂质体表面的zeta电位绝对值,从而进一步避免脂质体间出现聚集现象。
表1
2、益生菌存活力测试
取实施例1、对照例1和对照例2中制备的功能化可食性膜样品用甲醇溶剂超声破乳后,释放出菌体细胞,然后用平板活菌计数法在39℃下厌氧培养48~72h后,计算平板上的菌落数。
如图4所示,所述实施例1中益生菌活菌数为6.4×108cfu/mL,而对照例1中脂质体未经添加β-CG和对照例2中益生菌脂质体未添加鱼精蛋白聚集体的活菌数基本不存在。说明益生菌脂质体经过鱼精蛋白聚集体保护后,可避免高温固膜热处理对益生菌的损伤作用。而对照例1和对照例2中由于分别未添加β-CG和鱼精蛋白,因此未能在益生菌脂质体表面形成鱼精蛋白聚集体,使其经过高温固膜热处理时对益生菌的活性产生了很大的影响。
3、模拟胃液消化测试
对制备得到的载有益生菌的功能化可食性膜进行了体外模拟。益生菌活菌数的检测方法:采用国标《GB/T 4789.35-2016食品安全国家标准食品微生物学检测乳酸菌检测》。
模拟胃液制备:向1L去离子水中加入氯化钠2g和0.26g胃蛋白酶,用1M的盐酸将溶液pH调至1~2以制备出模拟胃液。将实施例1、对照例3中制备的待测样品取1g加入到模拟胃液中,同时,以无脂质体包裹的益生菌可食性膜作为空白样品对照。并在37℃、180rpm的水浴摇床中恒温震荡孵育,并分别在60min、120min和180min时取出1mL的模拟胃液,梯度稀释后利用平板菌落计数法测定益生菌的存活率。
如图5所示,采用脂质体包埋益生菌并用鱼精蛋白纤维聚集保护后,益生菌在胃环境的存活率大幅提升。在胃消化终点(180min)时,所述实施例1益生菌存活率为96%,对照例3中未经鱼精蛋白纤维聚集保护的脂质体益生菌的存活率有32%,而既无脂质体包裹又无鱼精蛋白纤维聚集保护的益生菌存活率只有5%。这表明,采用脂质体对益生菌进行包裹后,其在胃液环境中的存活率可得到改善,而通过本发明制备的经鱼精蛋白纤维聚集保护的益生菌脂质体中益生菌的存活率得到进一步大幅度提升,实施例1中制备的样品经过高温热处理且对益生菌基本无损伤,鱼精蛋白纤维聚集对益生菌脂质体提供了一定的保护作用。
Claims (4)
1.一种载有益生菌的功能化可食性膜,其特征在于,以质量百分比计,原料组成包括:醇溶蛋白25-30%、壳聚糖15-20%、增塑剂1-3%、鱼精蛋白聚集体保护的益生菌脂质体0.1-0.5%,余量为溶剂;
所述的鱼精蛋白聚集体保护的益生菌脂质体的制备方法包括如下步骤:
(1)、称取类脂、β-胆固醇-D-葡萄糖苷和抗氧化剂在有机溶剂中溶解,蒸发除去溶剂,形成脂质薄膜,随后在真空下干燥过夜;
(2)、将益生菌培养物悬浮于磷酸盐缓冲液中,得到细菌溶液;
(3)、将步骤(2)中的溶液加入到步骤(1)中脂质薄膜中,水合并涡旋形成脂质体混悬液,经高压均质后得到益生菌脂质体悬浮液;
(4)、将鱼精蛋白粉末溶于去离子水中,之后与步骤(3)得到的益生菌脂质体悬浮液混合均匀,搅拌孵育,制备得鱼精蛋白聚集体保护的益生菌脂质体;
所述类脂为两性离子脂质;所述抗氧化剂为VE乙酸酯、没食子酸丙酯中的任一种;
抗氧化剂用量为脂质体质量的0.5-1%;类脂和β-胆固醇-D-葡萄糖苷的摩尔比3.5-4:1,类脂和益生菌的质量比为6-15:1,类脂和鱼精蛋白的摩尔比1.3-2:1;
所述的载有益生菌的功能化可食性膜的制备方法包括如下步骤:
S1,将醇溶蛋白加入到乙醇溶液中,在热水浴中搅拌至溶解;
S2,将壳聚糖加入到冰醋酸中,在热水浴中搅拌至溶解;
S3,将上述溶液混合均匀后,加入增塑剂和鱼精蛋白聚集体保护的益生菌脂质体,充分搅拌,于热水浴中交联反应一段时间,真空脱气消泡;
S4,将溶液浇注于洁净干燥的聚四氟乙烯成膜版上,进行热处理固膜,制得载有益生菌的功能化可食性膜。
2.根据权利要求1所述的载有益生菌的功能化可食性膜,其特征在于,步骤(3)中所述高压均质处理的参数条件是:300-800bar压力下,均质1-10次。
3.根据权利要求1所述的载有益生菌的功能化可食性膜,其特征在于,所述醇溶蛋白为玉米醇溶蛋白、小麦醇溶蛋白中的至少一种。
4.根据权利要求1所述载有益生菌的功能化可食性膜,其特征在于,所述S1中醇溶蛋白-乙醇溶液质量浓度为9-13%;所述S2中壳聚糖-醋酸溶液质量浓度为3-5%;热水浴温度均为40-50℃;所述S3中,交联反应条件为50℃-55℃,水浴中反应1-2h;所述S4中,热处理条件为90-130℃,处理时间为10-20min。
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