CN117099892A - 乳双歧杆菌sf-b21的抗疲劳发酵饮品及其制备工艺 - Google Patents
乳双歧杆菌sf-b21的抗疲劳发酵饮品及其制备工艺 Download PDFInfo
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Abstract
本发明涉及微生物技术领域,具体涉及一种乳双歧杆菌SF‑B21的抗疲劳发酵饮品及其制备工艺,该抗疲劳发酵饮品由被发酵底物经乳双歧杆菌SF‑B21发酵后与调味料混合调味制得,被发酵底物包括:净化水62~82份,党参提取物1~3份,黄精提取物1~3份,甘草提取物3~5份,枸杞子提取物1~3份,当归提取物1~3份,沙棘提取物1~3份,山药粉3~5份,红枣粉3~5份;乳双歧杆菌SF‑B21于2021年9月23日保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC No.23474。本发明利用乳双歧杆菌SF‑B21发酵制备的抗疲劳发酵饮品口感良好,具有显著的抗疲劳作用。
Description
技术领域
本发明涉及微生物技术领域,具体涉及一种乳双歧杆菌SF-B21的抗疲劳发酵饮品及其制备工艺。
背景技术
慢性疲劳综合征(chronic fatigue syndrome,CFS)是一组以严重疲劳为主要症状的症候群,其原因不明并持续存在或反复发作,且患者经充分休息后仍得不到缓解。目前西医理论大多从能量代谢、神经***活动失调、代谢组学等方面认识疲劳,从而确定其生理和生化途径的变化,但对疲劳的研究认识发展过程相对单一。目前,虽然有一些药品和生物制品已被证实具有缓解疲劳的作用,但是很多化学药物有兴奋大脑皮质的作用,会导致惊厥、精神紊乱、成瘾等不良反应,临床应用受到限制。近来既可以作为天然食品食用又能达到治病防病效果的药食同源中药受到了科学家们的关注。
专利CN115444922A公开了一种抗疲劳药食同源组合物及其应用,原料包括以下组分:熟普洱6~9份、人参3~6份、薏苡仁12~15份、桑叶3~6份、金银花3~6份、葛根3~6份、藿香1~9份、生甘草2~4份、生姜2~4份以及大枣2~4份,经过粉碎、煎煮、冷却、浓缩、烘干得到一种抗疲劳药食同源组合物,能够有效缓解疲劳。专利CN115381099A公开了一种抗疲劳、提高免疫的药食同源组合物及其制备方法,包括以下重量份的原料,牡蛎20~25份、人参18~23份、鹿鞭16~20份、玛咖15~18份、大枣13~15份、枸杞10~15份、黄精8~12份和蛹虫草6~10份,原料经过煎煮、浓缩、制粒、干燥得到成品,经过试验对比,该抗疲劳、提高免疫的药食同源组合物对心理疲劳和脑力疲劳的完全缓解率达到60%以上。然而上述药食同源组合物多为原料煎煮、浓缩、干燥制得,存在口感差、见效慢的问题。
发明内容
针对现有抗疲劳的药食同源组合物存在口感差、见效慢的技术问题,本发明提供一种乳双歧杆菌SF-B21的抗疲劳发酵饮品及其制备工艺。
第一方面,本发明提供一种乳双歧杆菌SF-B21的抗疲劳发酵饮品,由被发酵底物经乳双歧杆菌(Bifidobacterium lactis)SF-B21发酵后与调味料混合调味制得,被发酵底物包括如下重量份的组分:净化水62~82份,党参提取物1~3份,黄精提取物1~3份,甘草提取物3~5份,枸杞子提取物1~3份,当归提取物1~3份,沙棘提取物1~3份,山药粉3~5份,红枣粉3~5份;乳双歧杆菌SF-B21保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC No.23474,保藏日期为2021年9月23日,保藏地址为北京市朝阳区北辰西路1号院3号。
进一步的,被发酵底物包括如下重量份的组分:净化水72份,党参提取物2份,黄精提取物2份,甘草提取物4份,枸杞子提取物2份,当归提取物2份,沙棘提取物2份,山药粉4份,红枣粉4份。
进一步的,调味料包括如下重量份的组分:蜂蜜1~3份,桃浓缩汁3~5份。
进一步的,调味料包括如下重量份的组分:蜂蜜2份,桃浓缩汁4份。
本发明发酵饮品的被发酵底物及调味料具备如下功效:
党参:甘,平,补中益气,和胃生津,祛痰止咳,用于脾虚食少便溏,四肢无力;
黄精:补气养阴,健脾,润肺,益肾;
甘草:补脾益气,清热解毒,祛痰止咳,缓急止痛,调和诸药;
枸杞:甘,平,滋肾,润肺;
当归:补血活血,调经止痛,润肠通便;
沙棘:健脾消食,止咳祛痰,活血散瘀;
山药:健脾,补肺,固肾,益精;
红枣:味甘性温、归脾胃经,有补中益气、养血安神、缓和药性的功能;
蜂蜜:具有调补脾胃的功效。
第二方面,本发明还提供一种上述抗疲劳发酵饮品的制备工艺,包括如下步骤:
(1)首先将被发酵底物的各组分混合,得到混合浆料;
(2)将混合浆料输送至灭菌容器中,120℃高压灭菌,冷却至35℃后得到灭菌液;
(3)将乳双歧杆菌SF-B21接种到MRS液体培养基中,37℃下培养得到接种剂;
(4)将接种剂接种于灭菌液中,37℃下培养得到发酵液;
(5)将发酵液输送至高压喷雾破壁机中,进行破壁处理,得到破壁发酵液;
(6)破壁发酵液与调味料混合调味后,得到抗疲劳发酵饮品。
进一步的,步骤(2)中高压灭菌时间为30min。
进一步的,步骤(3)中乳双歧杆菌SF-B21的接种量为1%,步骤(4)中接种剂的接种量为1%。
进一步的,步骤(5)中高压喷雾破壁机的压力设置为60MPa。
本发明的有益效果在于:
本发明提供的乳双歧杆菌SF-B21具有强肠胃适应能力,被发酵底物组分皆为药食同源,无毒副作用。相比于市售的抗疲劳产品,本发明利用乳双歧杆菌SF-B21发酵制备的抗疲劳发酵饮品口感良好,经试验验证,具有显著的抗疲劳作用且见效更快。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,对于本领域普通技术人员而言,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。
图1是使用实施例1-3和对比例1-3饲喂小鼠对小鼠悬尾累积不动时间的影响结果图。
图2是使用实施例1-3和对比例1-3饲喂小鼠对小鼠游泳累积不动时间的影响结果图。
具体实施方式
为了使本技术领域的人员更好地理解本发明中的技术方案,下面将结合本发明实施例中的附图,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都应当属于本发明保护的范围。
本发明使用的MRS固体平板培养基包括以下组分:蛋白胨10g、牛肉粉5g、酵母粉4g、K2HPO4•7H2O 2g、柠檬酸三铵2g、葡萄糖20g、CH3COONa•3H2O 5g、MgSO4•7H2O 0.2g、MnSO4•4H2O 0.05g、吐温80 1mL、琼脂15.0g、蒸馏水1000mL。
本发明使用的MRS液体培养基包括以下组分:蛋白胨10g、牛肉粉5g、酵母粉4g、K2HPO4•7H2O 2g、柠檬酸三铵2g、葡萄糖20g、CH3COONa•3H2O 5g、MgSO4•7H2O 0.2g、MnSO4•4H2O 0.05g、吐温80 1mL、蒸馏水1000mL。
本发明使用的党参提取物、黄精提取物、甘草提取物、枸杞子提取物、当归提取物、沙棘提取物的制备方法为:将原料洗净后破碎成粗粒,用10倍量水煎煮2次,煎煮时间为3h和1h,合并两次滤液,静置2h后过滤得提取液。将提取液在真空度为-0.06-0.08Mpa浓缩至相对密度为1.10~1.15(70℃),然后进行喷雾干燥,最后粉碎成80目细粉即得提取物。
各物质粉的制备方法如下:
山药粉:选择光滑、无病斑、条形直顺的新鲜铁棍山药;去除泥污,在清水中清洗干净;用削皮刀或竹片刮去外表皮,并挖除黑色斑眼。将去皮后的山药,切成0.2~0.3cm厚的薄片。山药切片后立即浸入0.5%亚硫酸氢钠水溶液中进行变色处理,浸泡2~3h后捞出;将捞出的山药片用清水漂去药液和胶体,然后放入沸水锅中烫漂6~8min,注意不要煮烂,捞出后再清水漂去黏液。将烫漂后的山药片置60~65℃烘房或烘箱内烘20h。将烘干后的山药片用电磨加工成粉。
红枣粉:将挑选好的红枣清洗后进行预煮,预煮温度70~90℃,红枣与水的料液比为1:5,预煮时间30min,得到预煮红枣汁;红枣汁去核并打碎为浆状,得到红枣浆,然后加入0.01%的果胶酶进行酶解,酶解温度30℃,酶解时间80min,真空加热浓缩至原重量的20%,得到浓缩红枣浆,采用两级高压进行均质处理,均质红枣浆进行喷雾干燥即得红枣粉。
桃浓缩汁的制备方法为:将原料挑选、修整后在100℃沸水中短时浸泡,用刷皮机除去表皮。水蜜桃需开瓢机并去核,然后用切丁机切丁后预煮5min,输送至打浆机打成浆,采用卧螺机提取汁液,同时用真空机除去氧气,用两效真空蒸发器,物料浓缩到36%~40%,然后进行高温杀菌,冷却后即得桃浓缩汁。
下列实施例中使用的乳双歧杆菌SF-B21菌株,已于2021年9月23日送至中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)进行保藏,保藏编号为CGMCC No.23474,保藏地址为北京市朝阳区北辰西路1号院3号,分类命名:Bifidobacterium lactis,其相关信息已公开于专利CN114668151A。
实施例1一种用乳双歧杆菌SF-B21制备的发酵饮品
(1)准备发酵饮品的原料,包括被发酵底物和调味料,被发酵底物包括如下重量份的组分:净化水72份,党参提取物2份,黄精提取物2份,甘草提取物4份,枸杞子提取物2份,当归提取物2份,沙棘提取物2份,山药粉4份,红枣粉4份;调味料包括如下重量份的组分:蜂蜜2份,桃浓缩汁4份。
(2)按照如下工艺步骤制备发酵饮品:
首先将被发酵底物的各组分混合,得到混合浆料;将混合浆料输送至灭菌容器中,120℃高压灭菌30min,然后冷却至35℃,得到灭菌液;将乳双歧杆菌SF-B21接种到MRS液体培养基中,接种量为1%,37℃下培养18h得到接种剂;将接种剂接种于灭菌液中,接种量为1%,37℃下培养24h,得到发酵液;将发酵液输送至高压喷雾破壁机中,压力设置为60MPa,进行破壁处理,得到破壁发酵液;破壁发酵液与调味料混合调味后,得发酵饮品。
实施例2一种用乳双歧杆菌SF-B21制备的发酵饮品
(1)准备发酵饮品的原料,包括被发酵底物和调味料,被发酵底物包括如下重量份的组分:净化水82份,党参提取物1份,黄精提取物1份,甘草提取物3份,枸杞子提取物1份,当归提取物1份,沙棘提取物1份,山药粉3份,红枣粉3份;调味料包括如下重量份的组分:蜂蜜1份,桃浓缩汁3份。
(2)按照实施例1中的工艺制备发酵饮品。
实施例3一种用乳双歧杆菌SF-B21制备的发酵饮品
(1)准备发酵饮品的原料,包括被发酵底物和调味料,被发酵底物包括如下重量份的组分:净化水62份,党参提取物3份,黄精提取物3份,甘草提取物5份,枸杞子提取物3份,当归提取物3份,沙棘提取物3份,山药粉5份,红枣粉5份;调味料包括如下重量份的组分:蜂蜜3份,桃浓缩汁5份。
(2)按照实施例1中的工艺制备发酵饮品。
对比例1 一种用两歧双歧杆菌CICC 10395制备的发酵饮品
(1)准备发酵饮品的原料,同实施例1。
(2)制备发酵饮品,与实施例1中的工艺不同之处在于:发酵菌种采用两歧双歧杆菌CICC 10395,其它工艺同实施例1。
对比例2 一种用动物双歧杆菌 CICC 21711制备的发酵饮品
(1)准备发酵饮品的原料,同实施例1。
(2)制备发酵饮品,与实施例1中的工艺不同之处在于:发酵菌种采用动物双歧杆菌CICC 21711,其它工艺同实施例1。
对比例3 一种抗疲劳维生素保健饮料
按照专利CN103211259B公开内容,制备一种抗疲劳维生素保健饮料,由以下原料制成:以1000kg总量为准,枸杞子提取物0.5kg、牛磺酸0.1kg、维生素B20.001kg、维生素B60.01kg、烟酰胺0.01kg,苹果酸0.1kg、柠檬酸1kg、柠檬酸钠1kg、白砂糖50kg、食用香精0.1kg,余量为水。其制备工艺如下:
(1)将枸杞子提取物在85℃水中搅拌20min溶解,制成枸杞子提取物溶液;(2)将牛磺酸、维生素B2、维生素B6、烟酰胺在65℃的水中预混搅拌,制成混合物溶液;(3)将枸杞子提取物溶液和混合物溶液泵入预混罐混合后加入苹果酸、柠檬酸、柠檬酸钠、白砂糖,搅拌溶解,经200目纱布管道过滤后泵入定容罐,添加食用香精进行调配、定容,再经121℃条件下杀菌15s后,冷却至95℃灌装,冷却,制成抗疲劳维生素保健饮料。
测试例1 乳双歧杆菌SF-B21的胃肠液耐受性测试
(1)模拟胃液的配置:125mmol/L NaCl、7mmol/L KCl、45mmol/L NaHCO3和3g/L胃蛋白酶,用盐酸调节pH至2.5,制成模拟胃液,以0.22μm无菌滤膜过滤,现配现用。
(2)模拟肠液的配置:45mmol/L NaCl、1g/L胰蛋白酶、3g/L牛胆盐,用氢氧化钠调节pH至8.0,制成模拟肠液,以0.22μm无菌滤膜过滤,现配现用。
(3)分别将两歧双歧杆菌CICC 10395、动物双歧杆菌CICC 21711和乳双歧杆菌SF-B21连续活化三代(每代活化时间为18h),测定连续活化三代后得到菌株在A600下的OD值,通过计算求出OD值=5所需的菌液量。
(4)将OD5的菌液以8000G离心10min,弃上清,重悬于1mL模拟胃液中,37℃培养3h后进行平板活菌计数;
(5)取模拟胃液处理后的菌液8000G离心10min,弃上清,重悬于等体积的模拟肠液中,37℃培养4h后进行平板活菌计数。
按照如下公式计算各菌株的耐受胃肠液后的存活率,分析各菌株的胃肠道适应能力,结果如表1所示。
表1 乳双歧杆菌SF-B21胃肠道适应性测试结果
如表1中的结果所示,与两歧双歧杆菌CICC 10395、动物双歧杆菌CICC 21711相比,乳双歧杆菌SF-B21具有更强的胃肠适应能力。
测试例2发酵饮品对小鼠疲劳性的影响测试
1、试验处理
将70只ICR小鼠随机分为7组,每组10只,按照下述方法饲喂:
对照组:正常饲喂,每天口服生理盐水5mL一次,连续饲喂28天;
实施例1组:正常饲喂,每天口服实施例1制备的发酵饮品5mL一次,连续饲喂28天;
实施例2组:正常饲喂,每天口服实施例2制备的发酵饮品5mL一次,连续饲喂28天;
实施例3组:正常饲喂,每天口服实施例3制备的发酵饮品5mL一次,连续饲喂28天;
对比例1组:正常饲喂,每天口服对比例1制备的发酵饮品5mL一次,连续饲喂28天;
对比例2组:正常饲喂,每天口服对比例2制备的发酵饮品5mL一次,连续饲喂28天;
对比例3组:正常饲喂,每天口服对比例3制备的饮料5mL一次,连续饲喂28天。
2、试验方法
(1)小鼠悬尾试验:在离小鼠尾尖部大概1cm处用胶贴上丝线一端,丝线另一端连接二导记录仪的张力换能器,小鼠头部离台面约7cm,悬挂总时长为6min,用二导记录仪记录2min后小鼠累积不动时间,小鼠呈倒悬垂状态,停止挣扎,静止不动为不动时间,结果如表2和图1所示。
(2)小鼠强迫游泳实验:将小鼠放入装满水的塑料桶内,保持水温恒定25℃,用数码相机记录下小鼠游泳的过程,总共6min,记录2min后小鼠累积不动时间,其中小鼠不再挣扎或漂浮于水面上、四肢轻微动作以保持头部能露出水面即可记录为不动时间,结果如表2所示和图2所示。
表2 发酵饮品对小鼠疲劳性的影响测试结果
组别 | 悬尾累计不动时间(s) | 游泳累计不动时间(s) |
对照组 | 71.69±16.28 | 68.23±15.23 |
实施例1组 | 44.35±10.21* | 45.18±10.02* |
实施例2组 | 58.27±12.23* | 60.22±15.26* |
实施例3组 | 55.33±13.23* | 59.18±16.39* |
对比例1组 | 60.18±15.19 | 65.19±20.16 |
对比例2组 | 64.36±10.57 | 63.20±16.72 |
对比例3组 | 55.21±12.14* | 54.32±13.33* |
注:*表示与对照组相比,p<0.05。
由以上图表看出,通过服用实施例1、实施例2、实施例3制备的发酵饮品,能够减少小鼠悬尾不动时间及游泳累计不动时间,与对照组具有显著性差异,说明乳双歧杆菌SF-B21制备的发酵饮品能够起到良好的抗疲劳作用。对比例3是市面上常见的富含维生素的抗疲劳饮料,其抗疲劳效果与对照组相比具有显著性差异,但是效果略差于实施例1,说明实施例1的抗疲劳效果优于市面上常见的抗疲劳维生素保健饮料。
尽管通过参考附图并结合优选实施例的方式对本发明进行了详细描述,但本发明并不限于此。在不脱离本发明的精神和实质的前提下,本领域普通技术人员可以对本发明的实施例进行各种等效的修改或替换,而这些修改或替换都应在本发明的涵盖范围内/任何熟悉本技术领域的技术人员在本发明揭露的技术范围内,可轻易想到变化或替换,都应涵盖在本发明的保护范围之内。
Claims (8)
1. 一种乳双歧杆菌SF-B21的抗疲劳发酵饮品,其特征在于,由被发酵底物经乳双歧杆菌(Bifidobacterium lactis)SF-B21发酵后与调味料混合调味制得,被发酵底物包括如下重量份的组分:净化水62~82份,党参提取物1~3份,黄精提取物1~3份,甘草提取物3~5份,枸杞子提取物1~3份,当归提取物1~3份,沙棘提取物1~3份,山药粉3~5份,红枣粉3~5份;乳双歧杆菌SF-B21保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCCNo.23474,保藏日期为2021年9月23日,保藏地址为北京市朝阳区北辰西路1号院3号。
2.如权利要求1所述的抗疲劳发酵饮品,其特征在于,被发酵底物包括如下重量份的组分:净化水72份,党参提取物2份,黄精提取物2份,甘草提取物4份,枸杞子提取物2份,当归提取物2份,沙棘提取物2份,山药粉4份,红枣粉4份。
3.如权利要求1所述的抗疲劳发酵饮品,其特征在于,调味料包括如下重量份的组分:蜂蜜1~3份,桃浓缩汁3~5份。
4.如权利要求3所述的抗疲劳发酵饮品,其特征在于,调味料包括如下重量份的组分:蜂蜜2份,桃浓缩汁4份。
5.一种如权利要求1所述抗疲劳发酵饮品的制备工艺,其特征在于,包括如下步骤:
(1)首先将被发酵底物的各组分混合,得到混合浆料;
(2)将混合浆料输送至灭菌容器中,120℃高压灭菌,冷却至35℃后得到灭菌液;
(3)将乳双歧杆菌SF-B21接种到MRS液体培养基中,37℃下培养得到接种剂;
(4)将接种剂接种于灭菌液中,37℃下培养得到发酵液;
(5)将发酵液输送至高压喷雾破壁机中,进行破壁处理,得到破壁发酵液;
(6)破壁发酵液与调味料混合调味后,得到抗疲劳发酵饮品。
6.如权利要求5所述的制备工艺,其特征在于,步骤(2)中高压灭菌时间为30min。
7.如权利要求5所述的制备工艺,其特征在于,步骤(3)中乳双歧杆菌SF-B21的接种量为1%,步骤(4)中接种剂的接种量为1%。
8.如权利要求5所述的制备工艺,其特征在于,步骤(5)中高压喷雾破壁机的压力设置为60MPa。
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