CN110679661A - 一种含具有降低血脂功效多糖的酸奶制备方法 - Google Patents
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Abstract
本发明目的是生产一种含具有降血脂功能细菌纤维素的酸奶,属于食品加工技术领域。本发明通过组织捣碎匀浆机得到Gluconacetobacter intermedius的细菌纤维素混悬液后灭菌杀灭Gluconacetobacter intermedius,再通过图1的步骤,即可获得本发明的酸奶。此产品特点在于富含活性多糖,这些多糖能够调节人体的血脂。实验过程中发现在含有无活性的Gluconacetobacter intermedius但富含有其代谢产物细菌纤维素的酸奶中,其降血脂的能力是最强的。
Description
技术领域
本发明涉及一种具有降血脂作用多糖与乳酸菌共同发酵制备酸奶的方法,属于食品加工技术领域。
背景技术
酸奶作为一种口味极佳,且能够改善肠道环境和促进消化吸收的饮品,它有着极大的市场,因此在这基础上,各公司为了增强酸奶的市场竞争力,筛选出一些能够对人体有益的菌,通过该菌发酵产生一系列的多糖。但是目前有报道指出,一位长期服用格列齐特的糖尿病患者,长期食用含益生菌的酸奶制品(干酪乳杆菌干酪亚种CNCM I-1518株),最终发现她的血液中含有与酸奶中的菌株相同的菌落,证实酸奶饮品并不适用于某些疾病的人群。上述病例的原因是该酸奶中除了用于酸奶发酵的发酵剂(嗜热链球菌和保加利亚乳杆菌)外还含有一种活性的干酪乳杆菌,而加入干酪乳杆菌(干酪亚种CNCM I-1518株)的目的在于它能够产生出一种具有抑制高血脂现象的多糖。
发明内容
为了解决这一问题,本发明找到一种能够高效产细菌纤维素的中间葡糖醋杆菌,在保留其发酵后的产物活性多糖后,对其灭菌,再将这些活性多糖与发酵剂共同发酵制备酸奶。
中间葡糖醋杆菌保藏于北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所的中国普通微生物菌种保藏管理中心,保藏号为CGMCC NO.8318,保藏日期为2013年10月10日,分类命名为Gluconacetobacter intermedius。
本发明目的是生产一种含具有降血脂功能细菌纤维素的酸奶,此产品特点在于富含活性多糖,这些多糖能够调节人体的血脂。目前发现CGMCC No.8318能够高效产细菌纤维素,而细菌纤维素作为一种多糖,是膳食纤维的一种,它们可以帮助降低慢性疾病的风险,如糖尿病、肥胖、心血管疾病和憩室炎。
本发明提供了一种将CGMCC No.8318产的细菌纤维素与发酵剂共同发酵的方法,包括以下步骤:
(1)筛选CGMCC No.8318
取10g醋醅装入混有玻璃珠和90mL富集培养基(酵母粉5g/L,蛋白胨3g/L,D-甘露醇25g/L,121℃灭菌20min)的500mL三角瓶中,30℃、200r/min过夜,菌悬液用无菌生理盐水10倍梯度稀释(10-1,10-2,10-3,10-4,10-5)。取200μL合适梯度稀释液涂布在分离培养基上(D-葡萄糖20g/L,蛋白胨8g/L,酵母粉5g/L,轻质碳酸钙7g/L,琼脂粉12g/L;pH6.8,121℃灭菌20min,冷却到50℃后加乙醇5mL/L)30℃培养3~5d。挑取具有透明圈的单菌落划线培养,得到纯菌。挑取单菌落在HS培养基(葡萄糖20g/L,蛋白胨5g/L,酵母粉5g/L,柠檬酸1.15g/L,磷酸氢二钠4g/L;pH 6.0、121℃灭菌20min,冷却后加无水乙醇20m L/L)中30℃、180r/min培养2d,菌株用终体积分数20%的甘油于-80℃保藏。
(2)CGMCC No.8318的发酵
将CGMCC No.8318接入种子培养液中,经两次转接至菌体密度≥5×108CFU/mL时作为种子液;将种子液按50~150mL接种量接入生产发酵培养液中后,置于200mm×250mm的浅层无菌托盘中至该液厚度为1.5~5cm,表面覆盖无菌纱布,培养室保持通风,发酵的条件为保持温度37℃,以甘油(4.3±0.11)g/L和葡萄糖(4.18±0.09)g/L为复合碳源,pH调节为6,发酵14d。
(3)将细菌纤维素同湿热链球菌、保加利亚乳杆菌发酵制备酸奶
将(2)中所得的菌膜即细菌纤维素凝胶取出,用清水洗净粘附于该凝胶下方的培养液残余物,将细菌纤维素膜在组织捣碎匀浆机中8000r/min破碎5min,4000r/min离心10min,收集并称量沉淀,加水制得1g/l00mL的细菌纤维素混悬液,在115℃下灭菌30min,将CGMCC No.8318灭活,后续步骤如图1。根据上述流程图最后得到本发明的酸奶。
本方法的特征在于先将CGMCC No.8318产的细菌纤维素灭菌,杀灭其上所有的CGMCC No.8318,接着将灭完菌的细菌纤维素同发酵剂(嗜热链球菌:保加利亚乳杆菌=1∶2)共同发酵得到酸奶。
附图说明
图1.流程图
图2.六组大鼠血清中甘油三酯和总胆固醇的含量关系图。
具体实施方式
以下结合优选实施例对本发明进行说明,应当理解此处所描述的优选实施例仅用于说明和解释本发明,并不用于限定本发明。
实施例1
本实施例的酸奶制备方法:将牛奶预热至50~55℃,以100mL牛奶为基础,加入100mg蔗糖,0.1%的细菌纤维素混悬液,将它们在90℃灭菌10min,再接种乳酸菌,发酵至酸度为70%后灭菌,此时的酸奶口感细腻,酸度甜度适中,具有酸奶独特香味。
实施例2
本实施例的酸奶制备方法,较实施例1相比,更改细菌纤维素含量为0.5%,蔗糖量为100mg,在90℃灭菌10min后,接种乳酸菌发酵至酸度为70%,再灭菌,此时的酸奶口感细腻,有淳厚感,具有酸奶独特香味,酸度甜度适中,适合大众口味。发酵结束后测定多糖含量,初始加入的细菌纤维素含量为实施例1的五倍,此时测得的多糖变化量较实施例2比也为5倍,该浓度无明显提升。
实施例3
本实施例的酸奶制备方法,较实施例1相比,更改细菌纤维素含量为1%,蔗糖量为100mg,在90℃灭菌10min后,接种乳酸菌发酵至酸度为70%,再灭菌,此时的酸奶的口感细腻,口味偏甜,具有酸奶独特香味。比较发现此实例中的细菌纤维素浓度为实施例1的10倍,而发酵所得酸奶多糖含量为实施例1组多糖含量的20倍,说明1%的细菌纤维素与发酵剂共同发酵能够促进多糖的产生。
实施例4
目前已有实验证明含汉氏葡糖酸醋杆菌(Gluconacetobacter hansenil)的酸奶能够起到降低总胆固醇和甘油三酯的功能,为了检测本发明的降血脂的功能,设计以下实验:
本实验采用高脂饮食结合链脲霉素建立Ⅱ型糖尿病大鼠模型,将大鼠分为ABCDEF五组,A组为正常大鼠,B组对糖尿病大鼠灌胃40%PBS甘油溶液,C组对糖尿病大鼠灌胃普通酸奶,D组对糖尿病大鼠灌胃含Gluconacetobacter hansenil的酸奶,E组对糖尿病大鼠灌胃含有CGMCC No.8318发酵产生的细菌纤维素但不含有该菌的酸奶,F组对糖尿病大鼠灌胃含CGMCC No.8318的酸奶,将六组大鼠在同一动物房内饲养,两个月后取血清检测总胆固醇和甘油三醇指标,结果如图2。
B组作为空白对照,C组灌胃普通酸奶其血脂指标较B组相似,酸奶本身并不会降低血脂。比较BDF两中含有产细菌纤维素菌的酸奶,发现两种酸奶都能起到降血脂的作用,但是含CGMCC No.8318的酸奶降血脂能力略强于含Gluconacetobacter hansenil的酸奶。比较BEF,EF两组的区别在于是否含有活性的CGMCC No.8318,结果表明,不含有活性的CGMCCNo.8318的酸奶,其降血脂的效果更加有效。
由上述实验结果表明使用本发明的技术手段,将发酵后的CGMCC No.8318灭菌,保留剩下的发酵产物同发酵剂共同发酵而成的酸奶,其降血脂的能力大于直接加入CGMCCNo.8318发酵而得的酸奶,也大于市面上已存在的含Gluconacetobacter hansenil的酸奶。
Claims (2)
1.一种含具有降低血脂功效多糖的酸奶制备方法,其特征为,采用中间葡糖醋杆菌(保藏于北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所的中国普通微生物菌种保藏管理中心,保藏号为CGMCC NO.8318,保藏日期为2013年10月10日,分类命名为Gluconacetobacter intermedius),进行发酵,步骤如下:
(1)CGMCC No.8318的发酵
将CGMCC No.8318接入种子培养液中,经两次转接至菌体密度≥5×108CFU/mL时作为种子液;将种子液按50~150mL接种量接入生产发酵培养液中后,置于200mm×250mm的浅层无菌托盘中至该液厚度为1.5~5cm,表面覆盖无菌纱布,培养室保持通风,发酵的条件为保持温度37℃,以甘油(4.3±0.11)g/L和葡萄糖(4.18±0.09)g/L为复合碳源,pH调节为6,发酵14d;
(2)将细菌纤维素同湿热链球菌、保加利亚乳杆菌发酵制备酸奶
将(1)中所得的菌膜即细菌纤维素凝胶取出,用清水洗净粘附于该凝胶下方的培养液残余物,将细菌纤维素膜在组织捣碎匀浆机中8000r/min破碎5min,4000r/min离心10min,收集并称量沉淀,加水制得1g/l00mL的细菌纤维素混悬液,在115℃下灭菌30min,将CGMCCNo.8318灭活,后续步骤如图1。根据上述流程图最后得到本发明的酸奶。
2.如权利要求1所述的方法,其特征为,先将CGMCC No.8318产的细菌纤维素灭菌,杀灭其上所有的CGMCC No.8318,接着将灭完菌的细菌纤维素同发酵剂共同发酵得到酸奶,其中发酵剂是以嗜热链球菌、保加利亚乳杆菌以质量比1∶2混合得到。
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