CN106579115B - 一种葡聚糖果蔬酵素的制备方法及应用 - Google Patents
一种葡聚糖果蔬酵素的制备方法及应用 Download PDFInfo
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Abstract
本发明属于食品保健领域,具体涉及一种葡聚糖果蔬酵素的制备方法及应用。所述制备方法包括以下步骤:(1)将果蔬清洗、晾干,粉碎,得混合果蔬液;(2)向混合果蔬液中加入糖,接种酵母菌,密封发酵,当pH<4.5时,得一次发酵液;(3)向一次发酵液中按重量百分比加入葡聚糖,接种复合乳酸菌进行二次发酵,复合乳酸菌包括植物乳杆菌、副干酪乳杆菌、干酪乳杆菌、醋酸杆菌。密封发酵,当pH<4.0时,得二次发酵液。(4)将二次发酵液过滤或离心,滤液/清液灭菌后,制得葡聚糖果蔬酵素。本发明葡聚糖果蔬酵素具有增强免疫力作用,可以开发成保健食品、功能食品和特殊医学用途食品,适合于免疫力低下人群长期的预防保健食用。
Description
技术领域
本发明属于食品保健领域,涉及一种果蔬酵素的制备方法及应用,特别涉及一种具有增强免疫力的酵素,具体涉及一种葡聚糖果蔬酵素的制备方法及应用。
背景技术
人们日常食用的水果和蔬菜中富含大量的非淀粉多糖类和多酚类成分,而对于大分子的非淀粉多糖和多酚类物质,由于人体缺乏相应的消化酶系,不能将之降解消化,只能通过肠道中的微生物来降解处理,产生对机体有益的代谢产物,如短链脂肪酸、多酚(原花青素)降解物等。这些代谢产物对人体有多种益处。然而,食物在肠道中停留的时间一般在24-48小时内,意味着肠道微生物对非淀粉类多糖和多酚类成分的降解作用时间非常有限,导致产生的有益代谢产物少,未能充分利用非淀粉类多糖和多酚类成分的全部益处。
酵素通常是指植物性的果蔬原料经微生物发酵后的产物,常含有一些具有活性的酶和果蔬成分的微生物代谢产物。不同的果蔬原料、微生物菌种、发酵时间的选择常导致酵素中功效性成分的差异,从而体现出对机体不同的功效作用和程度的差异。因此,在体外环境下利用微生物对果蔬的发酵处理,可以大大提升果蔬中有效成分对机体的生物学价值。随着社会的发展、生活环境的变化,人体免疫***受到越来越多的挑战,免疫力低下的状况如容易感冒、病后恢复慢、易过敏等情况越来越普遍。因此,开发源于天然食物、安全性高、适合长期服用的提升免疫力产品显得十分必要。
发明内容
为解决现有技术的缺点和不足之处,本发明的首要目的在于提供一种葡聚糖果蔬酵素的制备方法。
本发明的另一目的在于提供一种由上述制备方法得到的葡聚糖果蔬酵素。该酵素具有增强免疫力作用。
本发明的再一目的在于提供上述葡聚糖果蔬酵素的用途。
本发明目的通过以下技术方案实现:
一种葡聚糖果蔬酵素的制备方法,包括如下步骤:
(1)将果蔬清洗、晾干,切碎,然后粉碎成20~40目,得混合果蔬液;
(2)向步骤(1)得到的混合果蔬液中加入混合果蔬液重量百分比为1~10%的糖,接种混合果蔬液重量百分比0.01~0.05%的酵母菌,密封发酵,当pH<4.5时,得一次果蔬发酵液;
(3)向步骤(2)得到的一次果蔬发酵液中加入一次果蔬发酵液重量百分比0.1~1%的葡聚糖(β-glucan),按一次果蔬发酵液体积以100~1000CFU/ml接种复合乳酸菌进行二次发酵,所述复合乳酸菌包括植物乳杆菌、副干酪乳杆菌、干酪乳杆菌和醋酸杆菌;密封发酵,当pH<4.0时,得二次发酵液;
(4)将步骤(3)得到的二次发酵液混匀后过滤或离心,对滤液或离心上清液进行均质、灭菌后,制得葡聚糖果蔬酵素液。
优选的,如上所述的葡聚糖果蔬酵素制备方法中,在步骤(1)中,所述果蔬包括枇杷、梅子、菠萝、火龙果、木瓜、柑橘、桑椹、荔枝、龙眼、柠檬、梨、杨桃、葡萄、苹果、猕猴桃、草莓、樱桃、橙子、水蜜桃、西瓜、南瓜、胡瓜、白萝卜、胡萝卜、冬瓜、秋葵、花椰菜、芥兰、莴苣、甘蓝、青椒、西红柿和白菜中的多种。
优选的,在步骤(2)中加入的糖是蔗糖、葡萄糖和果葡糖浆中的一种或几种。
优选的,在步骤(2)中发酵温度为15~50℃,发酵时间为10~30天。
优选的,在步骤(3)中加入的葡聚糖(β-glucan)是香菇葡聚糖、茯苓葡聚糖和燕麦葡聚糖中的一种或几种。
优选的,在步骤(3)中所述混合菌的制备方法是植物乳杆菌、副干酪乳杆菌、干酪乳杆菌、醋酸杆菌等数量比例混合。
优选的,在步骤(3)中,所述发酵的条件为20~40℃,密闭发酵30~90天。
本发明还提供了上述方法制得的葡聚糖果蔬酵素在制备具有增强免疫力保健食品、功能食品或特殊医学用途食品中的应用。
通过常规制剂工艺、辅料和食品添加剂,上述葡聚糖果蔬酵素可以制成保健食品、功能食品和特殊医学用途食品中的多种剂型:软胶囊剂、硬胶囊、片剂、粉剂、颗粒剂、口服液、饮料、果冻或糖果。
与现有技术相比,本发明具有以下优点及有益效果:
本发明葡聚糖果蔬酵素的制备方法简单,制得的姜黄果蔬酵素源于天然食物、安全性高、适合长期服用,具有增强免疫力作用。
本发明制得的葡聚糖果蔬酵素能通过常规制剂工艺和辅料、食品添加剂,可以制成保健食品、功能食品和特殊医学用途食品中的多种剂型(如软胶囊剂、硬胶囊、片剂、粉剂、颗粒剂、口服液、饮料、果冻、糖果等),服用方便。
具体实施方式
下面结合实施例对本发明作进一步详细的描述,但本发明的实施方式不限于此。在未作说明的情况下,本发明涉及的原料与方法均为本领域的常规原料和方法。酵母菌Saccharomyces cerevisiae(购自安琪酵母)、植物乳杆菌Lactobacillus plantarum(购自上海交大昂立生物)、干酪乳杆菌lactobacillus casei(购自上海交大昂立生物)、副干酪乳杆菌Lactobacillus paracasei和醋酸杆菌Acetobacteraceti(购自广东省微生物研究所)。香菇葡聚糖和茯苓葡聚糖购自浙 江益圣菌物公司,燕麦葡聚糖购自西安锐博生物公司。
实施例1
(1)果蔬的清洗、破碎
将梅子、火龙果、木瓜、柠檬、梨、苹果、橙子、西瓜、白萝卜、胡萝卜、冬瓜、西红柿洗净、去皮和核、晾干,用剪切式粉碎机破碎至20目,得混合果蔬液。
(2)一次发酵
加入混合果蔬液重量百分比2%的蔗糖和0.02%的酵母菌,混合搅拌均匀后,室温15~30℃,密封发酵15天,每天搅拌发酵液1小时,取发酵液测得pH为4.3,得一次果蔬发酵液。
(3)二次发酵
向一次果蔬发酵液中加入一次果蔬发酵液重量百分比0.2%的茯苓葡聚糖,再按发酵液体积以100CFU/ml接入复合乳酸菌进行第二发酵,复合乳酸菌由植物乳杆菌、副干酪乳杆菌、干酪乳杆菌、醋酸杆菌等数量比例混合配制。室温20~30℃,密封发酵70天后,每天搅拌发酵液1小时,取发酵液测得pH为3.5,得二次发酵液。
(4)葡聚糖果蔬酵素液获得
将二次发酵液用离心机分离,以1000g离心力离心10分钟,取上清液均质、500ml玻璃瓶罐装灭菌后,制得葡聚糖果蔬酵素制品。
实施例2
(1)果蔬的清洗、破碎
将梅子、菠萝、火龙果、木瓜、柑橘、桑椹、杨桃、葡萄、苹果、草莓、橙子、水蜜桃、西瓜、白萝卜、胡萝卜、冬瓜、秋葵、花椰菜、西红柿、白菜洗净、去皮和核、晾干,用剪切式粉碎机破碎至20目,得混合果蔬液。
(2)一次发酵
加入混合果蔬液重量百分比5%的葡萄糖和0.05%的酵母菌,混合搅拌均匀 后,室温25~35℃,密封发酵10天,取发酵液测得pH为4.2,得一次果蔬发酵液。
(3)二次发酵
向一次果蔬发酵液加入一次果蔬发酵液重量百分比0.5%的燕麦葡聚糖,再按发酵液体积以1000CFU/ml接入复合乳酸菌进行第二发酵,复合乳酸菌由植物乳杆菌、副干酪乳杆菌、干酪乳杆菌、醋酸杆菌等数量比例混合配制。室温在25~35℃,密封发酵50天,每天搅拌一小时,取发酵液测得pH为3.4,得二次发酵液。
(4)葡聚糖果蔬酵素液获得
将二次发酵液用离心机分离,以1000g离心力离心10分钟,取上清液过均质机、500ml玻璃瓶罐装灭菌后,制得葡聚糖果蔬酵素制品。
实施例3
(1)果蔬的清洗、破碎
将枇杷、火龙果、木瓜、柑橘、杨桃、草莓、橙子、水蜜桃、西瓜、南瓜、白萝卜、胡萝卜、冬瓜、秋葵、花椰菜、芥兰、甘蓝、青椒、西红柿洗净、去皮和核后,晾干,用剪切式粉碎机破碎至30目,得混合果蔬液。
(2)一次发酵
加入混合果蔬液重量百分比10%的果葡糖浆和0.05%的酵母菌,混合搅拌均匀后,室温25~40℃,密封发酵20天,每天搅拌发酵液1小时,取发酵液测得pH为4.2,得一次果蔬发酵液。
(3)二次发酵
向一次果蔬发酵液中加入一次果蔬发酵液重量百分比1.0%的香菇葡聚糖,再按发酵液体积以1000CFU/ml接入复合乳酸菌进行第二发酵,复合乳酸菌由植物乳杆菌、副干酪乳杆菌、干酪乳杆菌、醋酸杆菌等数量比例混合配制。保温25~40℃,密封发酵90天,每天搅拌发酵液1小时,90天后取发酵液测得pH为3.3,得二次发酵液。
(4)葡聚糖果蔬酵素获得
将二次发酵液用离心机分离,以1000g离心力离心10分钟,取上清液均质、500ml玻璃瓶罐装灭菌后,制得葡聚糖果蔬酵素制品。
实施例4
(1)果蔬的清洗、破碎
将枇杷、火龙果、木瓜、柑橘、杨桃、草莓、橙子、水蜜桃、西瓜、南瓜、白萝卜、胡萝卜、冬瓜、秋葵、花椰菜、芥兰、甘蓝、青椒、西红柿洗净、去皮和核后,晾干,用剪切式粉碎机破碎至40目,得混合果蔬液。
(2)一次发酵
加入混合果蔬液重量百分比4%的果葡糖浆和4%的蔗糖,接种混合果蔬液重量百分比0.05%的酵母菌,混合搅拌均匀后,室温20~35℃,密封发酵30天,每天搅拌发酵液1小时,取发酵液测得pH为4.2,得一次果蔬发酵液。
(3)二次发酵
向一次果蔬发酵液中加入一次果蔬发酵液重量百分比0.1%的香菇葡聚糖,再按发酵液体积以1000CFU/ml接入复合乳酸菌进行第二发酵,复合乳酸菌由植物乳杆菌、副干酪乳杆菌、干酪乳杆菌、醋酸杆菌等数量比例混合配制。室温20~35℃,密封发酵30天,每天搅拌发酵液1小时,30天后取发酵液测得pH为3.5,得二次发酵液。
(4)葡聚糖果蔬酵素液获得
将二次发酵液用100目板框过滤机过滤,取滤液均质、500ml玻璃瓶罐装灭菌后,制得葡聚糖果蔬酵素制品。
实施例5
(1)果蔬的清洗、破碎
将火龙果、木瓜、柑橘、杨桃、草莓、橙子、西瓜、南瓜、白萝卜、胡萝卜、冬瓜、花椰菜、芥兰、甘蓝、西红柿洗净、去皮和核后,晾干,用剪切式粉碎机破碎至40目,得混合果蔬液。
(2)一次发酵
加入混合果蔬液重量百分比3%的蔗糖和3%的葡萄糖,接种混合果蔬液重量百分比0.02%的酵母菌,混合搅拌均匀后,室温25~40℃,密封发酵12天,每天搅拌发酵液1小时,取发酵液测得pH为4.1,得一次果蔬发酵液。
(3)二次发酵
向一次果蔬发酵液中加入一次果蔬发酵液重量百分比0.2%的香菇葡聚糖和0.3%的茯苓葡聚糖,再按发酵液体积以400CFU/ml接入复合乳酸菌进行第二发酵,复合乳酸菌由植物乳杆菌、副干酪乳杆菌、干酪乳杆菌、醋酸杆菌等数量比例混合配制。室温25~35℃,密封发酵60天,每天搅拌发酵液1小时,60天后取发酵液测得pH为3.3,得二次发酵液。
(4)葡聚糖果蔬酵素获得
将二次发酵液用100目板框过滤机过滤,取滤液均质、500ml玻璃瓶罐装灭菌后,制得葡聚糖果蔬酵素制品。
葡聚糖果蔬酵素的增强免疫力效果验证实验:
实验1:对小鼠迟发型***反应(DTH)的作用
动物及剂量设计:SPF级NIH系小鼠,体重18~22g,雌雄各半,每组10只。正常组(同体积生理盐水),模型组(同体积生理盐水),以实施例1、2、3、4、5制得的葡聚糖果蔬酵素为试验样品,试验剂量为5ml/kg·d,分别记为S1、S2、S3、S4、S5组。小鼠给药体积按0.2ml/10g计。试验方法按照***发《保健食品检验与评价技术规范》(2003年版)中要求。
数据处理:按统计学方法分析和处理数据,组间差异比较采用t检验。
结果与正常组比较,模型对照组的耳肿胀度显著下降(P<0.01),表明免疫低下造模成功。与模型组比较,各实施例组耳肿胀度显著升高(P<0.05-0.01)。提示各样品对免疫功能低下小鼠T细胞免疫功能具有增强作用。试验结果详见表1。
注:与正常组比较,##P<0.01;与模型组比较,*P<0.05,**p<0.01
实验2:对小鼠NK细胞杀伤活性的作用
供试品、剂量设置、分组同实验1。试验方法按照***发《保健食品检验与评价技术规范》(2003年版)中要求。
数据处理:按统计学方法分析和处理数据,组间差异比较采用t检验。
结果与正常组比较,模型组NK细胞的杀伤活性均显著降低(P<0.05),表明免疫功能低下动物造模成功。与模型组比较,各实施例样品组NK细胞的杀伤活性显著提高(P<0.05),提示各实施例样品后对动物的非特异性免疫功能具有提升作用。详见表2。
注:与正常对照组比较,πP<0.05;与模型对照组比较,*P<0.05
以上实验数据表明葡聚糖果蔬酵素具有增强免疫力的作用。以葡聚糖果蔬酵素为核心成分,可以开发成适合免疫力低下人群作为预防保健的长期服用的产品。
上述实施例为本发明较佳的实施方式,但本发明的实施方式并不受上述实施例的限制,其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化,均应为等效的置换方式,都包含在本发明的保护范围之内。
Claims (5)
1.一种葡聚糖果蔬酵素的制备方法,其特征在于,包括如下步骤:
(1)将果蔬清洗、晾干,切碎,然后粉碎成20~40目,得混合果蔬液;所述果蔬包括枇杷、梅子、菠萝、火龙果、木瓜、柑橘、桑椹、荔枝、龙眼、柠檬、梨、杨桃、葡萄、苹果、猕猴桃、草莓、樱桃、橙子、水蜜桃、西瓜、南瓜、胡瓜、白萝卜、胡萝卜、冬瓜、秋葵、花椰菜、芥兰、莴苣、甘蓝、青椒、西红柿和白菜中的多种;
(2)向步骤(1)得到的混合果蔬液中加入混合果蔬液重量百分比为1~10%的糖,接种混合果蔬液重量百分比0.01~0.05%的酵母菌,密封发酵,当pH<4.5时,得一次果蔬发酵液;所述发酵温度为15~50℃,发酵时间为10~30天;
(3)向步骤(2)得到的一次果蔬发酵液中加入一次果蔬发酵液重量百分比0.1~1%的葡聚糖,按一次果蔬发酵液体积以100~1000CFU/ml接种复合乳酸菌进行二次发酵,所述复合乳酸菌为等数量比例混合的植物乳杆菌、副干酪乳杆菌、干酪乳杆菌和醋酸杆菌;密封发酵,当pH<4.0时,得二次发酵液;所述葡聚糖是香菇葡聚糖、茯苓葡聚糖和燕麦葡聚糖中的一种或几种;所述发酵的条件为20~40℃,密闭发酵30~90天;
(4)将步骤(3)得到的二次发酵液混匀后过滤或离心,对滤液或离心上清液进行均质、灭菌后,制得葡聚糖果蔬酵素液。
2.根据权利要求1所述的一种葡聚糖果蔬酵素的制备方法,其特征在于,在步骤(2)中加入的糖是蔗糖、葡萄糖和果葡糖浆中的一种或几种。
3.一种葡聚糖果蔬酵素,其特征在于,其由权利要求1或2所述的一种葡聚糖果蔬酵素的制备方法制得。
4.如权利要求3所述的葡聚糖果蔬酵素在制备具有增强免疫力保健食品、功能食品或特殊医学用途食品中的应用。
5.根据权利要求4所述的应用,其特征在于,所述保健食品、功能食品和特殊医学用途食品的剂型是软胶囊剂、硬胶囊、片剂、粉剂、颗粒剂、口服液、饮料、果冻或糖果。
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