CN105962375A - 一种猴头菇活性乳酸菌制剂及其制备方法 - Google Patents
一种猴头菇活性乳酸菌制剂及其制备方法 Download PDFInfo
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- A—HUMAN NECESSITIES
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Abstract
本发明了公开了一种猴头菇活性乳酸菌制剂及其制备方法,所述制剂由猴头菇多糖、活性乳酸菌粉、益生元和辅料组成,具体配方为:猴头菇多糖25~30份,活性乳酸菌粉25~30份、益生元1~5份,辅料50~60份。本发明还提供了上述制剂的制备方法,步骤如下:(1)猴头菇多糖的制备;(2)活性乳酸菌粉的制备;(3)制备成品。该活性制剂同时具备猴头菇多糖对人体消化***的保健功能和多种乳酸菌联合发酵产生的有益活性菌群,加上利于肠道菌群生长繁殖的益生元,集三者合一,提供了一种养胃健胃的复合型制剂,该产品具有保护消化道粘膜,增强益生菌,抑制病原菌,标本兼治,治养兼得。产品携带及食用方便,适宜人群广泛。
Description
技术领域
本发明涉及一种保健品,具体是涉及一种以猴头菇多糖、活性乳酸菌粉和益生元,并辅以辅料制作而成的具有多重保健功能的一种制剂及其制备方法。
背景技术
多糖是一类具有三维立体结构的活性高分子化合物,是细胞受体的重要组成,在识别和传递信息,实现生物催化和物质转换的生理过程中起着非常重要的作用。猴头菇(Hericium erinaceu),自古以来就是著名的药食兼用真菌,不仅能烹制美味佳肴,又是辅助治疗医治消化不良,胃溃疡,十二指肠溃疡、食道癌、胃癌等消化***疾病的保健型食物,并具有很好的滋补作用。目前在国内外,猴头菇已经广泛被应用于医治消化道***的疾病与肿瘤。研究表明,猴头菇多糖是其主要的功效部位,是猴头菇的精华所在,具有:1)抑制肿瘤生长;2)促进消化道溃疡和各种慢性胃炎的修复;3)治疗肝炎;4)提高化疗后白细胞数量,调节机体免疫功能。临床使用结果已经证明,猴头菇多糖无任何毒、副作用。
活性乳酸菌是人体肠道正常菌群的主要微生物,对人体具有很多有益作用,包括排阻、抑制外来致病菌、提供维生素等营养,产生淀粉酶、蛋白酶等有助于消化的酶类,分解有毒或致癌物质(亚硝铵),产生有机酸、降低肠道pH和促进肠道的正常蠕动,刺激机体的免疫***并提高免疫力。活性乳酸菌在肠道的大量繁殖,可以治疗伪膜性肠炎(因大量使用抗生素而导致),治疗慢性腹泻和便秘,降低胆固醇,预防癌症的发生。
益生元是指能够选择性地促进人体肠道内原有的一种或几种有益细菌(益生菌)生长繁殖的物质,通过有益菌的繁殖增多,抑制有害细菌生长,从而达到调整肠道菌群,促进机体健康的目的。益生元不被人体消化和吸收,常见的有低聚木糖、乳果糖、蔗糖低聚糖、棉子低聚糖、异麦芽低聚糖、玉米低聚糖、大豆低聚糖等。
发明内容
本发明的目的是提供一种含有猴头菇提取物、活性乳酸菌和益生元的复合保健型制剂及其制备方法。
充分利用猴头菇多糖对胃肠道***疾病的预防和治疗作用,提取猴头菇精华部位,采用多种益生菌联合共发酵技术,人工干预引入肠道有益菌群,同时加入了大豆低聚糖和低聚木糖等有益于人体正常菌群增殖的益生元,为肠道细菌的持续增殖提供了物质基础。
本发明中一种猴头菇活性乳酸菌制剂,由猴头菇多糖、活性乳酸菌粉、益生元和辅料组成,所占重量份分别为:猴头菇多糖25~30份、活性乳酸菌粉25~30 份、益生元1~5份和辅料50~60份。
所述的猴头菇活性乳酸菌制剂,所述乳酸菌为嗜热链球菌,保加利亚乳酸杆菌、干酪乳杆菌、双歧杆菌,乳酸菌联合发酵剂为其中的两种、或三种、或多种组合,其接种量为乳酸菌发酵总体积的3~6%。
所述的猴头菇活性乳酸菌制剂,所述的益生元为大豆低聚糖和低聚木糖中的一种或是两种组合。
所述的猴头菇活性乳酸菌制剂的制备方法,所述的辅料为淀粉,可以是小麦淀粉,玉米淀粉,地瓜淀粉等,或是一种、或两种或多种组合。
所述的猴头菇活性乳酸菌制剂的制备方法,按照下述步骤进行:(1)猴头菇多糖的制备;(2)活性乳酸菌粉的制备;(3)制备成品。
所述的猴头菇活性乳酸菌制剂的制备方法,按照如下步骤制备猴头菇多糖:A.猴头原料的预处理;B.猴头菇多糖的溶出;C.超滤去除小分子;D.猴头菇多糖的醇沉;E.真空冷冻干燥;
A.原料的预处理:取干燥恒重的猴头菇实体,粉碎,置于8~16倍体积的60~95%乙醇溶液中浸泡18~30 h(优选的,置于9~12倍体积的80~95%乙醇溶液中浸泡20~24 h,更加优选的,置于10倍体积的95%乙醇溶液中浸泡22 h),以除去脂肪、部分单(寡)糖及脂溶性色素,残渣阴凉处风干;
B. 猴头菇多糖的溶出:预处理后的残渣风干后,选择提取时间为1~2 h,温度为60~100℃,水(蒸馏水)料比为20~30 mL/g(v/w),80~120目纱布过滤,滤渣重复提取2~3次;
C. 超滤去除小分子:将前面所述的多次提取的多糖溶液合并,采用超滤膜分子量为1万的中空纤维素膜进行超滤,去除1万以下的小分子物质;
D. 猴头菇多糖的醇沉:在超滤浓缩后的提取液中加无水乙醇,直至溶液的乙醇终点浓度为60~90%(优选的,加无水乙醇直至溶液终点浓度为75~85%,更加优选的,加无水乙醇直至溶液终点浓度为80%),4℃静置24 h,离心,取沉淀部分,冷冻干燥;
E. 真空冷冻干燥:先将多糖样品放置于-20℃下3 h,然后转入-80℃下冷冻6 h以上,然后转入真空冷冻干燥机干燥,得猴头菇多糖。
所述的猴头菇活性乳酸菌制剂的制备方法,按照如下步骤制备活性乳酸菌粉:A.培养基制备与灭菌;B. 菌种的活化;C. 接种发酵;D. 冷冻干燥,制成粉剂;
A. 培养基制备与灭菌:
(1)增殖培养基(MRS培养基)配方为:蛋白胨10.0g;牛肉膏10.0g;酵母膏5.0g;柠檬酸氢二铵2.0g;葡萄糖20.0g;吐温80 1.0mL;乙酸钠5.0g;磷酸氢二钾2.0g;硫酸镁0.58g;硫酸锰0.25g;蒸馏水1000mL;将pH调整到6.2~6.6,在121℃下,灭菌20 min;
(2)菌种活化培养基:取健康奶牛产的鲜牛奶,离心(5000 rpm,15 min),纱布过滤得脱脂乳(或者脱脂乳奶粉,按照1∶7的比例用蒸馏水配制),分装于三角瓶中或装入发酵罐中,108℃下,灭菌15 min;
B. 菌种的活化:冰箱保藏的菌种接入菌种活化培养基中传代3~5次后菌种活力完全恢复;
C. 接种发酵:将活力完全恢复的菌种按发酵液体积3~6% 的接种量,接入到MRS培养基中,37℃~40℃下恒温静置培养8~14 h后,离心(4℃,8000~10000 rpm, 10 min), 弃上清,按照65~95 g/L的量加入脱脂奶粉,充分混匀,制成菌悬液;
D. 冷冻干燥,制成粉剂:将上述的菌悬液先置于4℃平衡1 h,然后放于-20℃下3 h,然后转入-80℃下冷冻6 h,再转入真空冷冻干燥机干燥,制得活性乳酸菌粉。
所述的猴头菇活性乳酸菌制剂的制备方法,按照如下步骤制备成品:分别称取猴头菇多糖、活性乳酸菌粉、益生元和辅料,经干法制成片剂。
本发明产品同时具备猴头菇多糖的营养保健作用、活菌制剂的益生作用和益生元的选择性增殖作用,集三者合一,三位一体,提供了一种养胃健胃、具有持续增殖作用的保健型复合制剂;除此之外,本发明的优良效果还表现在:
(1)不仅拓展了猴头菇的利用途径,提升了猴头菇的附加值,也增加了乳酸菌制剂的品种类型,目前市场上未见有该产品;
(2)本发明产品携带及食用方便,适宜人群广泛,尤其对消化不良、食欲低下、便秘等胃肠功能紊乱者,以及免疫力低下、生活节奏紧张、饮食没有规律、易患胃肠疾病的职业人群,效果更佳,具有良好的社会效益和经济效益。
具体实施方式
下面结合实施例和实验例详细说明本发明的技术方案,但保护范围不限于此。
实施例1 一种猴头菇活性乳酸菌制剂的制备方法,步骤如下:
(1)猴头菇多糖的制备:A. 猴头原料的预处理;B. 猴头菇多糖的溶出;C. 超滤去除小分子;D. 猴头菇多糖的醇沉;E. 真空冷冻干燥;
A.猴头原料的预处理:取干燥恒重的猴头子实体1 kg,粉碎,置于15倍体积的95%乙醇溶液中浸泡24 h,破坏细胞壁并除去脂肪、部分单(寡)糖及脂溶性色素,离心(3000 rpm,20min),残渣阴凉处风干;
B.猴头菇多糖的溶出:风干后的残渣,选择提取时间为2 h,温度为90℃,水(蒸馏水)料比为25 ml/g(v/w),120目纱布过滤,滤渣重复提取2次;
C.超滤去除小分子:将2次提取的多糖溶液合并,采用超滤膜分子量为1万的中空纤维素膜进行超滤,去除1万以下的小分子物质;
D猴头菇多糖的醇沉:将超滤后的提取液离心,取上清加无水乙醇,直至溶液终点浓度为80%,4℃静置24 h,离心,取沉淀部分,冷冻干燥;
E. 真空冷冻干燥:先将多糖样品放置于-20℃下3 h,然后转入-80℃下冷冻6 h以上,然后转入真空冷冻干燥机干燥,得猴头菇多糖;
(2)活性乳酸菌粉的制备
A. 培养基制备与灭菌:
(1)增殖培养基(MRS培养基)配方为:蛋白胨10.0g;牛肉膏10.0g;酵母膏5.0g;柠檬酸氢二铵2.0g;葡萄糖20.0g;吐温801.0mL;乙酸钠5.0g;磷酸氢二钾2.0g;硫酸镁0.58g;硫酸锰0.25g;蒸馏水1000mL; 将pH 调整到6.6,在121℃下,灭菌20 min;
(2)菌种活化培养基:取脱脂乳奶粉,按照1∶7的比例用蒸馏水配制,分装于1000 mL三角瓶中,108℃下,灭菌15 min;
B. 菌种的活化:冰箱保藏的菌种接入菌种活化培养基中传代5次后菌种活力完全恢复;
C. 接种发酵:将完全活化后的菌种配成按照一定质量比配成复合菌剂,其复合菌剂为干酪乳杆菌和嗜热链球菌的混合物,干酪乳杆菌∶嗜热链球菌=1:1(w/w),按发酵液体积5%的量将复合菌剂接入MRS培养基中,37℃下恒温静置培养12 h后,离心(4℃,10000 rpm,10min),弃上清,按照80 g/L的量加入脱脂奶粉,充分混匀,制成菌悬液;
D. 冷冻干燥,制成粉剂:将上述的菌悬液先置于4℃平衡1 h,然后放于-20℃下3 h,然后转入-80℃下冷冻6 h,再转入真空冷冻干燥机干燥,制得活性乳酸菌粉;
(3)制备成品
分别称取猴头菇多糖、活性乳酸菌粉、低聚木糖和玉米淀粉,所占重量份分别为:30份、30 份、2份和和55 份,经干法制成片剂。
实施例2 一种猴头菇活性乳酸菌制剂的制备方法,步骤如下:
(1)猴头菇多糖的制备:A. 猴头原料的预处理;B. 猴头菇多糖的溶出;C. 超滤去除小分子;D. 猴头菇多糖的醇沉;E. 真空冷冻干燥;
A.猴头原料的预处理:取干燥恒重的猴头子实体1 kg,粉碎,置于10倍体积的95%乙醇溶液中浸泡22 h,破坏细胞壁并除去脂肪、部分单(寡)糖及脂溶性色素,离心(3000 rpm,20min),残渣阴凉处风干;
B.猴头菇多糖的溶出:风干后的残渣,选择提取时间为1.5 h,温度为90℃,水(蒸馏水)料比为25 mL/g(v/w),100目纱布过滤,滤渣重复提取3次;
C.超滤去除小分子:将3次提取的多糖溶液合并,采用超滤膜分子量为1万的中空纤维素膜进行超滤,去除1万以下的小分子物质;
D猴头菇多糖的醇沉:将超滤后的提取液离心,取上清加无水乙醇,直至溶液终点浓度为80%,4℃静置24 h,离心,取沉淀部分,冷冻干燥;
E. 真空冷冻干燥:先将多糖样品放置于-20℃下3 h,然后转入-80℃下冷冻6 h以上,然后转入真空冷冻干燥机干燥,得猴头菇多糖;
(2)活性乳酸菌粉的制备
A. 培养基制备与灭菌:
(1)增殖培养基(MRS培养基)配方为:蛋白胨10.0g;牛肉膏10.0g;酵母膏5.0g;柠檬酸氢二铵2.0g;葡萄糖20.0g;吐温80 1.0mL;乙酸钠5.0g;磷酸氢二钾2.0g;硫酸镁0.58g;硫酸锰0.25g;蒸馏水1000mL;将pH调整到6.6,在121℃下,灭菌20 min;
(2)菌种活化培养基:取健康奶牛产的鲜牛奶,离心(5000 rpm,15 min),纱布过滤得脱脂乳,分装于三角瓶中或装入发酵罐中,108℃下,灭菌15 min;
B. 菌种的活化:冰箱保藏的菌种接入菌种活化培养基中传代5次后菌种活力完全恢复;
C. 接种发酵:将完全活化后的菌种配成按照一定质量比配成复合菌剂,按发酵液体积5% 的量接入到MRS培养基中,其复合菌剂为保加利亚乳酸杆菌、嗜热链球菌和双歧杆菌的混合物,保加利亚乳酸杆菌∶嗜热链球菌∶双歧杆菌=6∶1∶1(w/w),37℃下恒温静置培养12h后,离心(4℃,10000 rpm, 10 min), 弃上清,按照80 g/L的量加入脱脂奶粉,充分混匀,制成菌悬液;
D. 冷冻干燥,制成粉剂:将上述的菌悬液先置于4℃平衡1 h,然后放于-20℃下3 h,然后转入-80℃下冷冻6 h,再转入真空冷冻干燥机干燥,制得活性乳酸菌粉;
(3)制备成品
分别称取猴头菇多糖、活性乳酸菌粉、大豆低聚糖和玉米淀粉,所占重量份分别为:28份、28 份、2份和和55 份,经干法制成片剂。
试验例1 本发明所得一种猴头菇活性乳酸菌制剂的抑菌试验:
选取大肠杆菌、沙门氏菌、金黄色葡萄球菌及溶血性链球菌等肠道细菌为实验菌种,将上述菌种接种于普通琼脂培养基上,37℃培养24 h,挑取典型菌落接种于新鲜肉汤培养基中,37℃培养18 h,备用;在无菌操作条件下,提取50 μL各菌液涂布于普通琼脂培养基上,并标记好所加菌种的名称,然后等距离打孔,孔直径为6 mm;分别将实施例1及实施例2所得片剂碾碎,加水制成浓度为1 g/mL的溶液,分别吸取100 μL两种溶液加入至孔洞内,放置于37℃恒温箱内培养24 h,观察并记录孔周围的抑菌圈直径(mm)。
抑菌效果判定:按照文献记载,抑菌圈直径≥20 mm为极敏;15~19 mm为高敏;10~14 mm为中敏;10 mm以下为低敏。
结果如表1所示:
表1 两种溶液对肠道细菌的抑菌作用:
由表1可知,本发明所得产品对上述四种肠道细菌的抑菌圈直径达到10.5~16 mm,其中对沙门氏菌及金黄色葡萄球菌的抑菌作用更好;且经对比可知,实施例2的抑菌圈直径均高于实施例1,说明实施例2的抑菌作用最突出。
应当指出的是,具体实施方式只是本发明比较有代表性的例子,显然本发明的技术方案不限于上述实施例,还可以有很多变形。本领域的普通技术人员,以本发明所明确公开的或根据文件的书面描述毫无异议的得到的,均应认为是本专利所要保护的范围。
本发明的完成由山东省自然基金资助 (No. ZR2015CL008)。
Claims (8)
1.一种猴头菇活性乳酸菌制剂,其特征在于,由猴头菇多糖、活性乳酸菌粉、益生元和辅料组成,所占重量份分别为:猴头菇多糖25~30份、活性乳酸菌粉25~30 份、益生元1~5份和辅料50~60份。
2.根据权利要求1 所述的猴头菇活性乳酸菌制剂,其特征在于,所述乳酸菌粉为嗜热链球菌、保加利亚乳酸杆菌、干酪乳杆菌、双歧杆菌,乳酸菌联合发酵剂为其中的两种、或三种、或多种组合,其接种量为乳酸菌发酵总体积的3~6%。
3.根据权利要求1 所述的猴头菇活性乳酸菌制剂,其特征在于,所述的益生元为大豆低聚糖和低聚木糖中的一种或是两种组合。
4.根据权利要求1 所述的猴头菇活性乳酸菌制剂的制备方法,其特征在于,所述的辅料为淀粉,可以是小麦淀粉,玉米淀粉,地瓜淀粉等,或是一种、或两种或多种组合。
5.根据权利要求1 所述的猴头菇活性乳酸菌制剂的制备方法,其特征在于,按照下述步骤进行:(1)猴头菇多糖的制备;(2)活性乳酸菌粉的制备;(3)制备成品。
6.根据权利要求5所述的猴头菇活性乳酸菌制剂的制备方法,其特征在于,按照如下步骤制备猴头菇多糖:A.猴头原料的预处理;B.猴头菇多糖的溶出;C.超滤去除小分子;D.猴头菇多糖的醇沉;E.真空冷冻干燥;
A.原料的预处理:取干燥恒重的猴头菇实体,粉碎,置于8~16倍体积的60~95%乙醇溶液中浸泡18~30 h(优选的,置于9~12倍体积的80~95%乙醇溶液中浸泡20~24 h,更加优选的,置于10倍体积的95%乙醇溶液中浸泡22 h),以除去脂肪、部分单(寡)糖及脂溶性色素,残渣阴凉处风干;
B. 猴头菇多糖的溶出:预处理后的残渣风干后,选择提取时间为1~2 h,温度为60~100℃,水(蒸馏水)料比为20~30 mL/g(v/w),80~120目纱布过滤,滤渣重复提取2~3次;
C. 超滤去除小分子:将前面所述的多次提取的多糖溶液合并,采用超滤膜分子量为1万的中空纤维素膜进行超滤,去除1万以下的小分子物质;
D. 猴头菇多糖的醇沉:在超滤浓缩后的提取液中加无水乙醇,直至溶液的乙醇终点浓度为60~90%(优选的,加无水乙醇直至溶液终点浓度为75~85%,更加优选的,加无水乙醇直至溶液终点浓度为80%),4℃静置24 h,离心,取沉淀部分,冷冻干燥;
E. 真空冷冻干燥:先将多糖样品放置于-20℃下3 h,然后转入-80℃下冷冻6 h以上,然后转入真空冷冻干燥机干燥,得猴头菇多糖。
7.根据权利要求5 所述的猴头菇活性乳酸菌制剂的制备方法,其特征在于,按照如下步骤制备活性乳酸菌粉:A. 培养基制备与灭菌;B. 菌种的活化;C. 接种发酵;D. 冷冻干燥,制成粉剂;
A. 培养基制备与灭菌:
(1)增殖培养基(MRS培养基)配方为:蛋白胨10.0g;牛肉膏10.0g;酵母膏5.0g;柠檬酸氢二铵2.0g;葡萄糖20.0g;吐温80 1.0mL;乙酸钠5.0g;磷酸氢二钾2.0g;硫酸镁0.58g;硫酸锰0.25g;蒸馏水1000mL;将pH调整到6.2~6.6,在121℃下,灭菌20 min;
(2)菌种活化培养基:取健康奶牛产的鲜牛奶,离心(5000 rpm,15 min),纱布过滤得脱脂乳(或者脱脂乳奶粉,按照1∶7的比例用蒸馏水配制),分装于三角瓶中或装入发酵罐中,108℃下,灭菌15 min;
B. 菌种的活化:冰箱保藏的菌种接入菌种活化培养基中传代3~5次后菌种活力完全恢复;
C. 接种发酵:将活力完全恢复的菌种按发酵液体积3~6% 的接种量,接入到MRS培养基中,37℃~40℃下恒温静置培养8~14 h后,离心(4℃,8000~10000 rpm, 10 min), 弃上清,按照65~95 g/L的量加入脱脂奶粉,充分混匀,制成菌悬液;
D. 冷冻干燥,制成粉剂:将上述的菌悬液先置于4℃平衡1 h,然后放于-20℃下3 h,然后转入-80℃下冷冻6 h,再转入真空冷冻干燥机干燥,制得活性乳酸菌粉。
8.根据权利要求5 所述的猴头菇活性乳酸菌制剂的制备方法,其特征在于,按照如下步骤制备成品:分别称取猴头菇多糖、活性乳酸菌粉、益生元和辅料,经干法制成片剂。
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