CN111100803B - 用于培养耐酸性乳酸菌的方法及其应用 - Google Patents
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Abstract
本发明公开了一种用于培养耐酸性乳酸菌的方法,包含将耐酸性乳酸菌接种于一培养基,该培养基基本上由米酒粕组成,以及在一酸性条件下培养该耐酸性乳酸菌。另一方面,本发明揭示一种组合物,其通过包含以下步骤的程序制备:以如前述的方法培养耐酸性乳酸菌得一第一产物,自该第一产物中去除米酒粕残渣得一第二产物,以及将该第二产物离心后去除沉淀的耐酸性乳酸菌得该组合物。
Description
技术领域
本发明涉及一种用于培养耐酸性乳酸菌的方法,以及应用所述方法得到的组合物。
背景技术
目前产业界利用市售培养基,或是利用乳酸菌生长特性自行配制培养基,但是 由于微生物需要的营养成分多达数十种,包括氮源、碳源、维生素及矿物质等,每 次配制需要额外添加许多品项,且必须依据不同菌种调整不同配方,造成人力及物 力的耗费。
发明内容
在一方面,本发明提供一种用于培养耐酸性乳酸菌的方法,包含将耐酸性乳酸 菌接种于一培养基,该培养基基本上由米酒粕组成,以及在一酸性条件下培养该耐 酸性乳酸菌。
根据本发明,优选的,该酸性条件为pH 3.5-4.2。在本发明的一具体实施例中, 该酸性条件为约pH 3.8。
根据本发明,优选的,以重量计,该培养基至少90%由米酒粕组成,更优选为 至少95%由米酒粕组成。
在本发明的一具体实施例中,该耐酸性乳酸菌为乳酸戊糖片球菌L-11(Pediococcus pentosaceus L-11),分类命名为乳酸戊糖片球菌L-11(Pediococcuspentosaceus L-11),该菌种保藏在中国典型培养物保藏中心,地址在中国武汉,菌种 保藏编号为CCTCC NO:M 2018381,保藏时间为2018年6月19日。
另一方面,本发明提供一种组合物,其通过包含以下步骤的程序制备:以如上 述的方法培养耐酸性乳酸菌得一第一产物,自该第一产物中去除米酒粕残渣得一第 二产物,以及将该第二产物离心后去除沉淀的耐酸性乳酸菌得该组合物。
根据本发明的部分具体实施例,该组合物制备为一食品或饮品组合物。
又一方面,本发明提供一种如上述的组合物在制备用于治疗高血压的医药组合物中的用途。
应了解先前的一般描述及以下的详述两者皆仅为示例性及解释性且并不限制 本发明。
附图说明
本发明先前的概述以及以下详述在配合随附图式阅读时得以更佳地被了解。为说明本发明,在图式中显示目前较佳的实施例。
在图示中:
图1显示以米酒粕或MRS作为乳酸菌的培养基,菌数成长的比较。
图2显示经不同发酵时间取得的本发明的组合物对血管升压素转换酶的抑制活性百分比。
图3显示以本发明的组合物管喂自发性高血压大鼠(spontaneous hypertensionrats,SHR)21天后大鼠的收缩压变化。
具体实施方式
在一方面,本发明提供一种用于培养耐酸性乳酸菌的方法,包含将耐酸性乳酸 菌接种于一培养基,该培养基基本上由米酒粕组成,以及在一酸性条件下培养该耐 酸性乳酸菌。
根据本发明,优选的,该酸性条件为pH 3.5-4.2。在本发明的一具体实施例中, 该酸性条件为约pH 3.8。
在本发明的较佳具体实施例中,在耐酸性乳酸菌接种前,该培养基经灭菌、调 整pH值及/或添加碳源(例如,糖蜜及蔗糖)。
在本发明的一具体实施例中,采用高温灭菌,例如,100℃或更高的温度,110℃ 或更高的温度,或120℃或更高的温度,灭菌时间可为10分钟或更长的时间, 或15分钟或更长的时间。
根据本发明,以重量计,该培养基至少90%、至少91%、至少92%、至少93%、 至少94%、至少95%、至少96%、至少97%、至少98%、至少99%或100%由米酒 粕所组成。
在本发明的一具体实施例中,该耐酸性乳酸菌为乳酸戊糖片球菌L-11(Pediococcus pentosaceus L-11),该菌种保藏在中国典型培养物保藏中心,菌种保 藏编号为CCTCC NO:M 2018381。
另一方面,本发明提供一种组合物,其通过包含以下步骤的程序制备:以如上 述的方法培养耐酸性乳酸菌得一第一产物,自该第一产物中去除米酒粕残渣得一第 二产物,以及将该第二产物离心后去除沉淀的耐酸性乳酸菌得该组合物。
根据本发明的部分具体实施例,该组合物制备为一食品或饮品组合物。
又一方面,本发明提供一种如上述的组合物在制备用于治疗高血压的医药组合物中的用途。
在本发明的一具体实施例中,该用于治疗高血压的医药组合物含有一本发明的组合物,以及一医药上可接受的载体。
本发明通过以下实例进一步说明,其提供作为例示而不限制本发明。
实施例1:本发明组合物的制备
米酒粕以121℃灭菌15分钟,冷却后备用。调整米酒粕的pH值至约3.8。将 新鲜培养的耐酸性乳酸菌Pediococcus pentosaceus L-11,以5%(v/v)接种于米酒粕 (菌量为1x106CFU/mL)。加入除菌后的糖蜜(5%(v/v))及蔗糖(1%(v/v)) 以增加碳源,并于32℃、150rpm发酵培养48小时。相较于市售培养基MRS,菌 数成长的结果相当且略佳(图1)。进行分段式离心,第一段以1,000rpm离心去除 酒粕渣,第二段以10,000rpm离心取得上清液(去除沉淀的耐酸性乳酸菌),得本 发明的组合物。
实施例2:抗高血压活性试验
进行抑制血管升压素转换酶(angiotensin-converting enzyme,下称「ACE」)活性的体外试验,结果如图2所示,本发明的组合物抑制ACE的IC50值为8.92±065 mg/mL胜肽。此外,进行动物试验,每日以管喂灌食自发性高血压大鼠(spontaneous hypertensionrats,SHR)本发明的组合物21天,其血压数值与空白组间具有显著的 差异(图3)。
本领域技术人员将理解可对上述具体实施例进行改变而不背离其广泛的发明 概念。因此,应当理解的是,本发明不限于所揭示的特定具体实施例,而是意在涵 盖在如随附的权利要求书所界定的本发明的精神与范畴内的修改。
Claims (5)
1.一种用于培养耐酸性乳酸菌的方法,该耐酸性乳酸菌为乳酸戊糖片球菌L – 11(Pediococcus pentosaceus L-11),该菌种保藏在中国典型培养物保藏中心,菌种保藏编号为CCTCC NO:M 2018381,该方法包含将耐酸性乳酸菌接种于一培养基,该培养基至少90%由米酒粕组成;以及在一酸性条件下培养该耐酸性乳酸菌,其中该酸性条件为pH 3.5-4.2。
2.如权利要求1所述的方法,其中,以重量计,该培养基至少95%由米酒粕组成。
3.一种制备具抗高血压活性的组合物的方法,其包含以下步骤:
以权利要求1或2所述的方法培养耐酸性乳酸菌得一第一产物,该耐酸性乳酸菌为乳酸戊糖片球菌L – 11(Pediococcus pentosaceus L-11),该菌种保藏在中国典型培养物保藏中心,菌种保藏编号为CCTCC NO:M 2018381;
自该第一产物中去除米酒粕残渣得一第二产物;以及
将该第二产物离心后去除沉淀的耐酸性乳酸菌得该组合物。
4.按照权利要求3所述的方法制备得到的组合物在制备具抑制血管升压素转换酶活性的药物中的用途。
5.一种耐酸性乳酸菌,其为乳酸戊糖片球菌L – 11(Pediococcus pentosaceus L- 11),该菌种保藏在中国典型培养物保藏中心,菌种保藏编号为CCTCC NO:M 2018381。
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