CN111728199B - 一种牛樟芝酵素及其制备方法与应用 - Google Patents
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Abstract
本发明提供了一种牛樟芝酵素及其制备方法与应用,属于生物技术领域。第一方面,本发明提供一种牛樟芝酵素,牛樟芝酵素由牛樟芝、葡萄糖、人参药渣、益生菌、硫酸镁等组分制得;第二方面,本发明提供上述牛樟芝酵素的制备方法;第三方面,本发明还提供牛樟芝酵素的应用。利用本发明进行发酵得到的牛樟芝酵素水平与现有技术相比,均有显著提高。
Description
技术领域
本发明涉及生物技术领域,具体而言,涉及一种牛樟芝酵素及其制备方法与应用。
背景技术
牛樟芝,又名樟菇,属于非褶菌目、多孔科、多年生蕈菌类,学名为(Antrodiacamphorata)是1990年才被生化界发表的新种。
牛樟芝本身具有强烈之樟树香气,外形多变,有板状、钟状、马蹄状或塔状。一年生,无柄,木栓质至木质;紧贴着生于木材表面。表面褐色至黑褐色,具不明显的皱纹,有光泽,边缘平而钝。菌肉两层,上层木材色,下层象牙色,厚1~1.5 公分。表面孔状,微细绵密,每毫米有4 或5 个孔,管口小;初生时呈深红色,渐变为乳白色、淡褐色或淡黄褐色。子实体周边常呈放射反卷,并四周扩展生长,呈半圆形或不规则形。
牛樟芝中含有多种生理活性成分,如多糖、三萜类化合物、超氧歧化酶(SOD)腺苷、蛋白质(含免疫蛋白)、多种维生素、微量元素(钙、磷、楮)、核酸、凝集素、氨基酸、胆固醇、木质素、血压稳定物质等。其具备的生理活性功能具有抗肿瘤、增强免疫力、抗病毒、抗过敏、抗高血压、抑制血小板凝集、降血压、降胆固醇、抗细菌、保护肝脏等。
酵素是日文的叫法,是具有生物本身自然生成的具有催化功能的生物大分子—生物催化剂。据史书记载,我国3000多年前的就已经开始应用酶了,如酒曲。酵素存在于所有活的动植物体内,是维持机体正常功能,消化食物,修复组织等生命活动的一种必需物质。生物体内含有千百种酵素,它们支配着生物的新陈代谢、营养和能量转换等许多催化过程,与生命过程关系密切的反应大多是酵素催化反应。酵素能促进新陈代谢,使人精力充沛,心情愉悦;促进血液循环,同时净化血液,排除毒素;增强胃肠道的消化吸收功能,增强体质;调节人体酸碱平衡。
鉴于此,本发明提供一种利用牛樟芝菌丝体发酵制备牛樟芝酵素的方法。
发明内容
本发明的目的在于提供一种牛樟芝酵素及其制备方法与应用。
为了实现本发明的上述目的,特采用以下技术方案:
第一方面,本发明提供一种牛樟芝酵素产品,由牛樟芝、葡萄糖、人参药渣、益生菌、硫酸镁经发酵制得。
在本发明较佳的实施例中,按重量份数计,牛樟芝为0.2~1份、葡萄糖10~18份、人参药渣为5~12份、益生菌0.2~0.4份、硫酸镁为0.01~0.03份。
进一步地,在本发明较佳的实施例中,按重量份数计,牛樟芝为0.4~0.8份、葡萄糖12~16份、人参药渣为6~10份、益生菌0.2~0.4份、硫酸镁为0.01~0.03份和马铃薯冻干超微粉为5~8份。
进一步地,在本发明较佳的实施例中,按重量份数计,牛樟芝为0.6份、葡萄糖15份、人参药渣为8份、益生菌0.3份、硫酸镁为0.02份、马铃薯冻干超微粉为7份。
本发明中发酵所用牛樟芝为菌丝体冻干后经超微粉碎获得的粉体,马铃薯冻干超微粉为马铃薯粉碎后冻干后再经超微粉碎获得的粉体,人参药渣为人参经65%的酒精常规回流提取得到,其他组分在市场上购得。
第二方面,本发明提供上述牛樟芝酵素的制备方法,其包括:
牛樟芝0.2~1份、葡萄糖10~18份、人参药渣为5~12份、益生菌0.2~0.4份、硫酸镁为0.01~0.03份,加水量为上述物质重量份数总和的7~12倍,混匀后121℃灭菌15~30min后冷却。
进一步地,所述牛樟芝为0.4~0.8份、葡萄糖12~16份、人参药渣为6~10份、益生菌0.2~0.4份、硫酸镁为0.01~0.03份和马铃薯冻干超微粉为5~8份,加水量为上述物质重量份数总和的7~12倍,混匀后121℃灭菌15~30min后冷却。
进一步地,在本发明较佳的实施例中,所述牛樟芝为0.6份、葡萄糖15份、人参药渣为20份、益生菌0.3份、硫酸镁为0.02份和马铃薯冻干超微粉为7份,加水量为上述物质重量份数总和的7~12倍,混匀后121℃灭菌15~30min后冷却。
第三方面,本发明还提供牛樟芝酵素及其制备方法与应用,其包括:将益生菌接种至上述的发酵基质中,温度25~33℃,通气搅拌,发酵时间5~9d,将培养好的发酵液过滤或离心2000~10000rpm处理,上清液经40~50℃真空浓缩,即得牛樟芝酵素。
进一步地,在本发明较佳的实施例中,将益生菌接种至上述的发酵基质中,温度29℃,通气搅拌,发酵时间7d,将培养好的发酵液离心4000rpm处理,上清液经40~50℃真空浓缩,即得牛樟芝酵素。
该牛樟芝酵素与食品、特殊食品或药学上可接受的载体或辅料可以制成口服制剂。
该制剂可以是散剂、片剂、胶囊剂、颗粒剂、口服液。
该牛樟芝酵素在制备对胃黏膜损伤有保护作用的食品、特殊食品或药物中的应用。
与现有技术相比,本发明的有益效果包括:
牛樟芝的菌丝体干重和量与现有技术相比,均有显著提高。
具体实施方式
下面将结合实施例对本发明的实施方案进行详细描述,但是本领域技术人员将会理解,下列实施例仅用于说明本发明,而不应视为限制本发明的范围。实施例中未注明具体条件者,按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者,均为可以通过市售购买获得的常规产品。
以下结合实施例对本发明的特征和性能作进一步的详细描述:
实施例1
牛樟芝2g、葡萄糖100g、人参药渣120g、硫酸镁0.3g,加水量为上述物质重量份数总和的7~12倍,混匀后121℃灭菌30min后冷却。
将乳杆菌2g接种至上述的发酵基质中,温度33℃,通气搅拌,发酵时间5d,将培养好的发酵液离心10000rpm处理,上清液经40~50℃真空浓缩,即得牛樟芝酵素。
实施例2
牛樟芝10g、葡萄糖180g、人参药渣为50g、硫酸镁为0.1g,加水量为上述物质重量份数总和的7~12倍,混匀后121℃灭菌15min后冷却。
将酿酒酵母4g接种至上述的发酵基质中,温度25℃,通气搅拌,发酵时间5d,将培养好的发酵液过滤,上清液经40~50℃真空浓缩,即得牛樟芝酵素。
实施例3
牛樟芝为4g、葡萄糖160g、人参药渣为60g、硫酸镁为0.3g和马铃薯冻干超微粉为50g,加水量为上述物质重量份数总和的7~12倍,混匀后121℃灭菌20min后冷却。
将乳杆菌2g接种至上述的发酵基质中,温度25℃,通气搅拌,发酵时间9d,将培养好的发酵液离心2000rpm处理,上清液经40~50℃真空浓缩,即得牛樟芝酵素。
实施例4
牛樟芝为8g、葡萄糖120g、人参药渣为100g、硫酸镁为0.1g和马铃薯冻干超微粉为80g,加水量为上述物质重量份数总和的7~12倍,混匀后121℃灭菌20min后冷却。
将酿酒酵母4g接种至上述的发酵基质中,温度29℃,通气搅拌,发酵时间7d,将培养好的发酵液离心3000rpm处理,上清液经40~50℃真空浓缩,即得牛樟芝酵素。
实施例5
牛樟芝为6g、葡萄糖150g、人参药渣为80g、硫酸镁为0.2g和马铃薯冻干超微粉为70g,加水量为上述物质重量份数总和的7~12倍,混匀后121℃灭菌25min后冷却。
将酿酒酵母3g接种至上述的发酵基质中,温度29℃,通气搅拌,发酵时间7d,将培养好的发酵液离心4000rpm处理,上清液经40~50℃真空浓缩,即得牛樟芝酵素。
实施例6
本实施例提供一种胶囊剂,该片剂的制备方法包括:
上述实施例5牛樟芝酵素加入适量淀粉、糊精,混合均匀,制成颗粒,压片,包薄膜衣,即得片剂。
实施例7
本实施例提供一种胶囊剂,该胶囊剂的制备方法包括:
上述实施例5牛樟芝酵素加入适量淀粉、糊精,混匀,制成颗粒,装入硬胶囊,即得胶囊。
实施例8
本实施例提供一种颗粒剂,该颗粒剂的制备方法包括:
上述实施例5牛樟芝酵素加入适量淀粉、糊精,混匀,湿法制粒,干燥后整粒,包装,即得颗粒剂。
对比例1
牛樟芝为6g、葡萄糖为300g和硫酸镁为0.2g,加水量为上述物质重量份数总和的7~12倍,混匀后121℃灭菌25min后冷却。
将酿酒酵母3g接种至上述的发酵基质中,温度29℃,通气搅拌,发酵时间7d,将培养好的发酵液离心4000rpm处理,上清液经40~50℃真空浓缩,即得牛樟芝酵素。
对比例2
牛樟芝为6g、人参药渣为300g和硫酸镁为0.2g,加水量为上述物质重量份数总和的7~12倍,混匀后121℃灭菌25min后冷却。
将酿酒酵母3g接种至上述的发酵基质中,温度29℃,通气搅拌,发酵时间7d,将培养好的发酵液离心4000rpm处理,上清液经40~50℃真空浓缩,即得牛樟芝酵素。
对比例3
牛樟芝为6g、马铃薯冻干超微粉为300g、硫酸镁为0.2g,加水量为上述物质重量份数总和的7~12倍,混匀后121℃灭菌25min后冷却。
将酿酒酵母3g接种至上述的发酵基质中,温度29℃,通气搅拌,发酵时间7d,将培养好的发酵液离心4000rpm处理,上清液经40~50℃真空浓缩,即得牛樟芝酵素。
对比例4
未经冻干的牛樟芝菌丝体超微粉为6g、葡萄糖150g、人参药渣为80g、酿酒酵母3g、硫酸镁为0.2g和马铃薯冻干超微粉为70g,加水量为上述物质重量份数总和的7~12倍,混匀后121℃灭菌25min后冷却。
将益生菌接种至上述的发酵基质中,温度29℃,通气搅拌,发酵时间7d,将培养好的发酵液离心4000rpm处理,上清液经40~50℃真空浓缩,即得牛樟芝酵素。
本发明的有益效果:
实验例1
表1 活性成分检测结果
序号 | 组别 | 总黄酮含量 | SOD活性(U/g) | 蛋白酶活性(U/g) | 多酚含量 |
1 | 实施例1组 | 2.86% | 338 | 255 | 0.16% |
2 | 实施例2组 | 3.28% | 331 | 261 | 0.19% |
3 | 实施例3组 | 3.21% | 358 | 272 | 0.18% |
4 | 实施例4组 | 3.50% | 372 | 286 | 0.23% |
5 | 实施例5组 | 3.68% | 398 | 316 | 0.25% |
6 | 实施例6组 | 3.61% | 392 | 311 | 0.23% |
7 | 实施例7组 | 3.72% | 377 | 313 | 0.25% |
8 | 实施例8组 | 3.65% | 386 | 322 | 0.24% |
9 | 对比例1组 | 2.52% | 155 | 162 | 0.11% |
10 | 对比例2组 | 2.57% | 163 | 191 | 0.13% |
11 | 对比例3组 | 2.62% | 189 | 186 | 0.15% |
12 | 对比例4组 | 2.71% | 201 | 192 | 0.15% |
由表1可知,实施例1~8组与对比例1~4组相比,实施例1~8组特别是实施例5~8组活性成分检测结果具有明显优势。
实验例2
对乙醇致大鼠胃黏膜损伤的保护作用选取健康SD大鼠,雌雄各半,体质量180~220g,随机份为对照组、实施例1~8组、对比例1~5组,共13组,每组10只动物。每日灌胃给药1次,剂量均为2g/kg,连续7d,对照组给予等量淀粉。实验前,大鼠先禁食48h,自由饮水,于末次给药后1h每只大鼠灌胃无水乙醇1mL/只,再过1h脱颈椎处死动物,用夹子夹闭贲门,自幽门注入1%甲醛5mL,再夹闭幽门,放入1%甲醛溶液固定10min,沿胃大弯剪开胃壁,测量损伤长度,以损伤长度(宽度大于1mm加倍)总和作为胃黏膜损伤指数,并计算抑制率。
抑制率(%)=(对照组溃疡指数-给药组溃疡指数)/对照组溃疡指数×100%
表2 对乙醇所致大鼠急性胃黏膜损伤的影响实验结果(`x±s,n=10)
序号 | 组别 | 溃疡指数(mm) |
1 | 对照组 | 92.3±28.1 |
2 | 实施例1组 | 46.2±21.5 |
3 | 实施例2组 | 40.9±21.0 |
4 | 实施例3组 | 38.3±20.3 |
5 | 实施例4组 | 29.5±19.1 |
6 | 实施例5组 | 23.2±15.9 |
7 | 实施例6组 | 20.8±14.2 |
8 | 实施例7组 | 22.6±16.3 |
9 | 实施例8组 | 21.8±17.0 |
10 | 对比例1组 | 61.5±20.1 |
11 | 对比例2组 | 60.1±19.2 |
12 | 对比例3组 | 59.8±17.8 |
13 | 对比例4组 | 53.3±16.9 |
由表2可知,实施例1~8组与对照组及对比例1~4组相比,实施例1~8组特别是实施例5~8组对乙醇所致大鼠急性胃黏膜损伤的恢复程度具有明显优势。
本发明中未详细说明的环节均为本领域普通技术人员可自行选择的公知常识。虽然,上文中已经用一般性说明及具体实施方案对本发明作了详尽的描述,但在本发明基础上,可以对之作一些修改或改进,这对本领域普通技术人员而言是显而易见的。因此,在不偏离本发明精神的基础上所做的这些修改或改进,均属于本发明要求保护的范围。
Claims (8)
1.一种牛樟芝酵素,其特征在于,所述牛樟芝酵素在有氧发酵的条件下,由以下重量份数的成分制成:牛樟芝菌丝体冻干超微粉为0.4~0.8份、葡萄糖12~16份、人参药渣为6~10份、酿酒酵母0.2~0.4份、硫酸镁为0.01~0.03份和马铃薯冻干超微粉为5~8份;所述牛樟芝酵素的制备方法是将所述牛樟芝菌丝体冻干超微粉、所述葡萄糖、所述人参药渣、所述硫酸镁和所述马铃薯冻干超微粉,加水量为上述物质重量份数总和的7~12倍,混匀后121℃灭菌15~30min后冷却,即为酵素的发酵基质;然后将所述酿酒酵母接种至所述发酵基质中,温度25~33℃,通气搅拌,发酵时间5~9d,将培养好的发酵液过滤或离心2000~10000rpm处理,上清液经40~50℃真空浓缩,即得所述牛樟芝酵素。
2.根据权利要求1所述的牛樟芝酵素,其特征在于,按重量份数计,所述牛樟芝菌丝体冻干超微粉为0.6份、所述葡萄糖15份、所述人参药渣为8份、所述酿酒酵母0.3份、所述硫酸镁为0.02份和所述马铃薯冻干超微粉为7份。
3.根据权利要求1所述的牛樟芝酵素,其特征在于,所述马铃薯冻干超微粉为马铃薯粉碎后冻干后再经超微粉碎获得的粉体。
4.根据权利要求1所述的牛樟芝酵素,其特征在于,所述酿酒酵母活菌数≥1.0×107CFU/mL。
5.根据权利要求1所述的牛樟芝酵素,其特征在于,所述发酵温度为29℃;所述发酵时间为7d;所述发酵液离心速度为4000rpm。
6.根据权利要求5所述的牛樟芝酵素,其特征在于,所述牛樟芝酵素制成散剂、片剂、胶囊剂、颗粒剂或口服液。
7.一种如权利要求1~6所述的牛樟芝酵素在食品的应用。
8.一种如权利要求1~6所述的牛樟芝酵素在制备对胃黏膜损伤有保护作用的药物中的应用。
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