CN109170908A - 一种微胶囊益生菌粉及其制备方法 - Google Patents
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Abstract
本发明公开了属于食品加工技术领域的一种微胶囊益生菌粉及其制备的方法。该微胶囊益生菌粉以还原性多糖与蛋白的美拉德产物为微胶囊壁材包封益生菌,并经喷雾干燥形成粉末;美拉德产物在增强人体胃肠道功能、对人体肠道菌群产生益生作用的同时,还作为大分子物质保护益生菌,解决了益生菌在贮存过程中对外界不良环境敏感、容易死亡、必须贮存于冷藏条件下、对贮存条件要求高等缺点。
Description
技术领域
本发明属于食品加工技术领域,特别涉及一种微胶囊益生菌粉及其制备方法。
背景技术
益生菌具有调节肠道菌群、降低胆固醇、抑制有害细菌、抗过敏、提高机体免疫力等功能,是目前研究的热点。由于益生菌在贮存过程中对外界不良环境敏感、容易死亡、对贮存条件要求高,因此在贮存过程以及人体消化道条件下如何使益生菌保持活性发生益生作用是食品工业中亟需解决的问题。
近年来,微胶囊化益生菌引发研究者们广泛的兴趣。微胶囊益生菌的优点包括:可保护益生菌免受噬菌体的作用;提高益生菌冷冻和冷冻干燥的生存率;保证益生菌在贮藏中更好的稳定性等等。但是目前利用微胶囊保护益生菌的现有技术中,都未能解决益生菌通过胃肠屏障后仍能保持高活菌率的要求,且都需要冷冻或冷藏贮存,这样就对微胶囊化益生菌的生产、贮存、应用的条件提出了较高的要求,极大地限制了益生菌产品的工业生产、贮存、运输、销售环节的实际应用。
发明内容
本发明的目的在于提供一种微胶囊益生菌粉及其制备的方法,具体技术方案如下:
一种微胶囊益生菌粉以还原性多糖与蛋白的美拉德反应产物为微胶囊壁材包封益生菌芯材。
所述还原性多糖含有羰基,包括ι-卡拉胶和/或果胶;蛋白包括大豆分离蛋白、乳清蛋白、酪蛋白中的一种或多种。
所述益生菌为长双歧杆菌、短双歧杆菌、动物双歧杆菌、鼠李糖乳杆菌中的一种或多种。
所述微胶囊益生菌粉的制备方法包括以下步骤:
(1)将还原性多糖与蛋白以质量比为(1:3)~(3:1)混合,在相对湿度79%、温度60℃下反应12h~48h后,用去离子水溶解得到浓度为0.5%w/v的美拉德反应产物;
(2)制备益生菌菌悬液:将益生菌冻干菌粉活化后接种于MRS培养基,充氮后37℃厌氧培养24h,离心后倒出上清液,将得到的菌体沉淀以0.85%生理盐水清洗2次并重悬于生理盐水中,菌体浓度调整为2×109CFU mL-1;其中MRS培养基组成为:蛋白胨10g,牛肉膏10g,酵母膏5g,K2HPO4·3H2O 2g,乙酸钠5g,葡萄糖20g,吐温80 1mL,柠檬酸二铵2g,MgSO4·7H2O 0.58g,MnSO4·4H2O 0.25g,琼脂18g,加蒸馏水至1000mL,调节pH 6.2-6.4;
(3)将步骤(2)得到的益生菌菌悬液与步骤(1)得到的美拉德反应产物以体积比为1:(6~9)混合,将得到的混合溶液采用蠕动泵或连续恒流泵以6.25mL/min~12.50mL/min的流量泵入喷雾干燥设备进行喷雾干燥,喷雾干燥设备入口温度为110℃~130℃,出口温度为70℃~80℃,;收集喷干后的粉末即为微胶囊益生菌粉。
相比益生菌初始数量2×109CFU/g,上述微胶囊益生菌粉在20℃的贮藏条件下贮藏15d后,活菌数比初始菌数仅下降了0.3~2.0数量级,与相同贮藏条件下未经微胶囊保护的裸菌相比(活菌数比初始菌数下降2.5~3.5个数量级)表现出对益生菌良好的保护作用;经过模拟胃液、模拟肠液处理后,微胶囊菌粉中活菌数仅下降2个数量级,仍能达到维持肠道健康的国际标准。
本发明的有益效果为:
(1)本发明根据每一种益生菌表面疏水性、表面电位等特性,针对性地制备对应的美拉德反应产物作为相应益生菌的微胶囊壁材。芯材、壁材的体积比仅为1:(6~9),既节省了壁材的使用量,又能保证壁材对益生菌的保护作用;美拉德反应产物不仅增强了人体胃肠道功能、对人体肠道菌群的益生作用,还作为大分子物质保护益生菌,维持益生菌活性;解决了益生菌在贮存过程中对外界不良环境敏感、容易死亡、对贮存条件要求高等缺点;
(2)本发明制备得到的微胶囊益生菌粉抗逆性强,经过模拟胃液模拟肠液连续处理后的存活率高;在常温、冷藏条件下贮存较长时间仍能保持很高的活力,突破了益生菌产品只能冷链运输、贮藏、保质期短等技术瓶颈;
(3)本发明选择喷雾干燥,直接使溶液、乳浊液干燥成粉状或颗粒状制品,有效保持了益生菌活性且缩短了生产流程,制备方法简单高效;可以广泛应用在保健品领域,如制作成方便携带、独立包装的bar或添加在各种食品中,填补国内产品的空白,有着广阔的市场前景。
具体实施方式
本发明提供了一种微胶囊益生菌粉及其制备的方法,下面结合实施例对本发明做进一步的说明。
实施例1
按照下述步骤制备微胶囊益生菌粉:
(1)称取大豆分离蛋白1.639g、ι-卡拉胶4.918g,充分混合后置于相对湿度为79%、温度为60℃的条件下,经干法反应12小时后,加去离子水复溶美拉德反应产物,使其浓度为0.5%(w/v);
(2)将长双歧杆菌冻干菌粉活化后接种于MRS培养基,充氮后在37℃下厌氧培养24h,在4℃、6000r/min下离心10min,倾出上清液后,菌体沉淀以0.85%生理盐水清洗2次并重悬于生理盐水中,菌体浓度调整至2×109CFUmL-1;
(3)将步骤(2)得到的菌液与步骤(1)得到的美拉德产物溶液按照体积比1:7,将菌液添加到美拉德产物壁材中,搅拌均匀后,进行喷雾干燥:喷雾干燥条件为入口温度120℃,出口温度70℃,泵速为10.42mL/min;收集到的粉末即为微胶囊益生菌粉。
以0.1mL未微胶囊化的长双歧杆菌悬液作对照组,测试上述微胶囊益生菌存活率:
(1)4℃冷藏条件下:将上述喷雾干燥法制备的微胶囊益生菌、对照组细菌均置于4℃条件下储藏,分别测定第1d、3d、7d、15d微胶囊中的活菌数,以评价其保藏特性。微胶囊益生菌经喷雾干燥后活菌数的对数值为9.13,对照组经喷雾干燥后活菌数的对数值为8.85;微胶囊益生菌粉在4℃的贮藏条件下贮藏15d后,活菌数仍能达到5.6×108CFU mL-1,比初始菌数仅下降了0.38个数量级,而对照组的活菌数为7.14×107CFU mL-1,比初始菌数下降了1.01个数量级;
(2)常温贮藏条件下:将上述喷雾干燥法制备的微胶囊益生菌粉在20℃的贮藏条件下贮藏15d后,活菌数仍能达到2.20×108CFU mL-1,比初始菌数仅下降了0.79数量级,对照组的活菌数为1.07×106CFU mL-1,比初始菌数下降了2.82个数量级;
(3)微胶囊化的长双歧杆菌在连续的模拟胃肠液中的存活实验:分别称取0.1g实施例1制备的微胶囊益生菌、0.1mL对照组长双歧杆菌悬液,将其均置于0.9mL人工胃液中,37℃、100r/min的摇床上温育1h后,离心收集微胶囊,用生理盐水洗涤后加入0.9mL人工肠液,在60min和120min时取样进行活菌计数,平行3次取均值。
结果表明:经喷雾干燥处理使活菌数由添加量2×109CFU mL-1降至1.35×109CFUmL-1,而对照组降至7.10×108CFU mL-1,说明壁材在喷雾干燥的过程中对菌体表现出较好的保护性;微胶囊菌粉在4℃和20℃条件下贮存15天后活菌数仍有5.61×108CFU mL-1和2.20×108CFU mL-1,远高于对照组在这两种贮存条件下的活菌数,说明在冷藏和常温的贮存条件下微胶囊壁材能较好的保障益生菌的活菌数量;对比冷藏和常温两种贮存条件,虽然常温贮藏会对益生菌的存活率造成一定的影响,但20℃的贮存条件下,本法制备的微胶囊益生菌粉的存活率仍然很高,长期贮存实验表明,菌粉中的活菌数在3个月之后降至1.0×107CFU mL-1以下,经7个月之后降至1.0×106CFU mL-1以下;微胶囊益生菌粉经连续的模拟胃肠液处理后,仍有4.1×107CFU mL-1的活菌数存留,而对照组已检测不到活菌数。
实施例2
按照下述步骤制备微胶囊益生菌粉:
(1)称取大豆分离蛋白3.279g、果胶3.279g,充分混合后置于相对湿度为79%、温度为60℃的条件下,经干法反应24小时后,加去离子水复溶美拉德产物,使其浓度为0.5%(w/v);
(2)将短双歧杆菌冻干菌粉活化后接种于MRS培养基,充氮后在37℃下厌氧培养24h,在4℃、6000r/min下离心10min,倾出上清液后,菌体沉淀以0.85%生理盐水清洗2次并重悬于生理盐水中,菌体浓度调整至2×109CFUmL-1;
(3)将步骤(2)得到的菌液与步骤(1)得到的美拉德产物溶液按照体积比1:8,将菌液添加到美拉德产物壁材中,搅拌均匀后,进行喷雾干燥:喷雾干燥条件为入口温度130℃,出口温度80℃,泵速为12.50mL/min;收集到的粉末即为微胶囊益生菌粉。
以0.1mL未微胶囊化的短双歧杆菌悬液作对照组,测试上述微胶囊益生菌存活率:
(1)4℃冷藏:将上述喷雾干燥法制备的微胶囊益生菌、对照组细菌均置于4℃条件下储藏,分别测定第1d、3d、7d、15d微胶囊中的活菌数,以评价其保藏特性。微胶囊益生菌经喷雾干燥后活菌数的对数值为9.10,对照组经喷雾干燥后活菌数的对数值为8.25;在4℃的贮藏条件下贮藏15d后,活菌数仍能达到5.10×108CFU mL-1,比初始菌数仅下降了0.39个数量级,而对照组的活菌数为1.06×107CFU mL-1,比初始菌数下降了1.22个数量级;
(2)常温贮藏:将上述喷雾干燥法制备的微胶囊益生菌粉在20℃的贮藏条件下贮藏15d后,活菌数仍能达到9.12×107CFU mL-1,比初始菌数仅下降了1.14数量级,对照组的活菌数为8.07×105CFU mL-1,比初始菌数下降了2.34个数量级;
(3)微胶囊化的短双歧杆菌在连续的模拟胃肠液中的存活实验:分别称取0.1g实施例2制备的微胶囊益生菌、0.1mL对照组短双歧杆菌悬液,将其均置于0.9mL人工胃液中,37℃、100r/min的摇床上温育1h后,离心收集微胶囊,用生理盐水洗涤后加入0.9mL人工肠液,在60min和120min时取样进行活菌计数,平行3次取均值。
结果表明:经喷雾干燥处理使活菌数由添加量2×109CFU mL-1降至1.26×109CFUmL-1,而对照组降至1.78×108CFU mL-1,说明壁材在喷雾干燥的过程中对菌体表现出较好的保护性;微胶囊菌粉在4℃和20℃条件下贮存15天后活菌数仍有5.10×108CFU mL-1和9.12×107CFU mL-1,远高于对照组在这两种贮存条件下的活菌数,说明在冷藏和常温的贮存条件下微胶囊壁材能较好的保障益生菌的活菌数量;对比冷藏和常温两种贮存条件,虽然常温贮藏会对益生菌的存活率造成一定的影响,但20℃的贮存条件下,本法制备的微胶囊益生菌粉的存活率仍然很高;微胶囊益生菌粉经连续的模拟胃肠液处理后,仍有9.71×106CFU mL-1的活菌数存留,而对照组已检测不到活菌数。
实施例3
按照下述步骤制备微胶囊益生菌粉:
(1)称取乳清蛋白4.38g、ι-卡拉胶2.19g,充分混合后置于相对湿度为79%、温度为60℃的条件下,经干法反应36小时后,加去离子水复溶美拉德产物,使其浓度为0.5%(w/v);
(2)将鼠李糖乳杆菌冻干菌粉活化后接种于MRS培养基,充氮后在37℃下厌氧培养24h,在4℃、6000r/min下离心10min,倾出上清液后,菌体沉淀以0.85%生理盐水清洗2次并重悬于生理盐水中,菌体浓度调整至2×109CFUmL-1;
(3)将步骤(2)得到的菌液与步骤(1)得到的美拉德产物溶液按照体积比1:9,将菌液添加到美拉德产物壁材中,搅拌均匀后,进行喷雾干燥:喷雾干燥条件为入口温度110℃,出口温度80℃,泵速为8.34mL/min;收集到的粉末即为微胶囊益生菌粉。
以0.1mL未微胶囊化的鼠李糖乳杆菌悬液作对照组,测试上述微胶囊益生菌存活率:
(1)4℃冷藏:将上述喷雾干燥法制备的微胶囊益生菌、对照组细菌均置于4℃条件下储藏,分别测定第1d、3d、7d、15d微胶囊中的活菌数,以评价其保藏特性。微胶囊益生菌经喷雾干燥后活菌数的对数值为8.95,对照组经喷雾干燥后活菌数的对数值为8.80;在4℃的贮藏条件下贮藏15d后,活菌数仍能达到3.16×108CFU mL-1,比初始菌数仅下降了0.45个数量级,而对照组的活菌数为4.27×107CFU mL-1,比初始菌数下降了1.17个数量级;
(2)常温贮藏:将上述喷雾干燥法制备的微胶囊益生菌粉在20℃的贮藏条件下贮藏15d后,活菌数仍能达到4.51×107CFU mL-1,比初始菌数仅下降了1.31个数量级,对照组的活菌数为1.35×106CFU mL-1,比初始菌数下降了2.66个数量级;
(3)微胶囊化鼠李糖乳杆菌在连续的模拟胃肠液中的存活实验:分别称取0.1g实施例3制备的微胶囊益生菌、0.1mL对照组鼠李糖乳杆菌悬液,将其均置于0.9mL人工胃液中,37℃、100r/min的摇床上温育1h后,离心收集微胶囊,用生理盐水洗涤后加入0.9mL人工肠液,在60min和120min时取样进行活菌计数,平行3次取均值。
结果表明:经喷雾干燥处理使活菌数由添加量2×109CFU mL-1降至8.90×108CFUmL-1,而对照组降至6.31×108CFU mL-1,说明壁材在喷雾干燥的过程中对菌体表现出较好的保护性;微胶囊菌粉在4℃和20℃条件下贮存15天后活菌数仍有3.16×108CFU mL-1和4.5×107CFU mL-1,远高于对照组在这两种贮存条件下的活菌数,说明在冷藏和常温的贮存条件下微胶囊壁材能较好的保障益生菌的活菌数量;对比冷藏和常温两种贮存条件,虽然常温贮藏会对益生菌的存活率造成一定的影响,但20℃的贮存条件下,本法制备的微胶囊益生菌粉的存活率仍然很高;微胶囊益生菌粉经连续的模拟胃肠液处理后,仍有9.36×106CFUmL-1的活菌数存留,而对照组未能检测出活菌。
Claims (8)
1.一种微胶囊益生菌粉,其特征在于,以还原性多糖与蛋白的美拉德反应产物为微胶囊壁材包封益生菌芯材。
2.根据权利要求1所述的微胶囊益生菌粉,其特征在于,所述还原性多糖含有羰基,包括ι-卡拉胶和/或果胶。
3.根据权利要求1所述的微胶囊益生菌粉,其特征在于,所述蛋白包括大豆分离蛋白、乳清蛋白、酪蛋白中的一种或多种。
4.根据权利要求1所述的微胶囊益生菌粉,其特征在于,所述益生菌为长双歧杆菌、短双歧杆菌、动物双歧杆菌、鼠李糖乳杆菌中的一种或多种。
5.权利要求1~4任一项所述微胶囊益生菌粉的制备方法,其特征在于,所述方法包括以下步骤:
(1)还原性多糖与蛋白在相对湿度79%、温度60℃下反应12h~48h后,用去离子水溶解得到浓度为0.5%w/v的美拉德反应产物;
(2)混合益生菌菌悬液与美拉德产物,喷雾干燥后得到微胶囊益生菌粉。
6.根据权利要求5所述的制备方法,其特征在于,所述步骤(1)中还原性多糖与蛋白的质量比为(1:3)~(3:1),步骤(2)中益生菌与美拉德产物的体积比为1:(6~9)。
7.根据权利要求5所述的制备方法,其特征在于,所述步骤(2)中益生菌菌悬液的制备为:将益生菌冻干菌粉活化后接种于MRS培养基,充氮后37℃厌氧培养24h,离心后倒出上清液,将得到的菌体沉淀以0.85%生理盐水清洗2次并重悬于生理盐水中,菌体浓度调整为2×109CFU mL-1。
8.根据权利要求5所述的制备方法,其特征在于,所述步骤(2)中喷雾干燥设备入口温度为110℃~130℃,出口温度为70℃~80℃,进料流量为6.25mL/min~12.50mL/min。
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