CN115210359A - 利用红参促进乳酸菌增殖的培养工艺 - Google Patents
利用红参促进乳酸菌增殖的培养工艺 Download PDFInfo
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- CN115210359A CN115210359A CN202180018655.XA CN202180018655A CN115210359A CN 115210359 A CN115210359 A CN 115210359A CN 202180018655 A CN202180018655 A CN 202180018655A CN 115210359 A CN115210359 A CN 115210359A
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- lactic acid
- acid bacteria
- lactobacillus
- red ginseng
- culturing
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Abstract
本发明提供了:一种包括红参来源的膳食纤维的乳酸菌培养基,以及一种使用该培养基培养乳酸菌的方法。根据本发明,在乳酸菌培养基中使用红参来源的膳食纤维,从而乳酸菌的活细胞数可以增加、且可培养性可以提高。另外,本发明的培养乳酸菌的方法,使用该培养基,从而在相同条件下提高了可培养性,且因此在成本方面存在优势。
Description
技术领域
本发明涉及一种包含红参原料的乳酸菌用培养基及使用该培养基培养乳酸菌的方法。
背景技术
人参(Panax ginseng C.A.Meyer)作为五加科人参属的多年生草药,是东方医学中使用的草药之一,其味甘苦、具有温补的能量,并具有如强元气、保脾和健肺等草药功效。
人参的主要生理活性物质包含皂苷(saponin)(也称为人参皂苷(ginsenoside))、精油成分、聚乙炔、酚成分、糖苷、酸性肽等,并且包含另外各种成分例如维生素、糖、和矿物质。据报道,普通人参的生理活性功效具有对中枢神经系统的效果、增强免疫功能的效果、抗癌效果等。特别是,迄今为止,已经发现了约40种类型的人参皂苷,并且已经证实其通过广泛影响内分泌系统、免疫系统、代谢系统等包含中枢神经系统而显示出对机体功能调节即生理功能正常化的优异效果。这些人参皂苷具有相似或相反的作用,并且已知单独通过特定成分或通过其几种类型的相互作用而表现出各种作用。
另一方面,膳食纤维是已知作为在食品中的蔬菜、水果、海草中含有的大量纤维或纤维素等的成分,并且是不被人的消化酶消化而排出体外的高分子碳水化合物。已知纤维通过使胃感觉不太饥饿并延迟食物的吸收以过滤胆固醇而作为膳食中的有益成分。这些纤维富含于蔬菜的韧性部分(纤维素)、萝卜的棕色素(木质素)、水果中的果胶、以及海芥和海带,尤其是蘑菇,的粘性成分(海藻酸)。
乳酸菌是产生大量乳酸作为代谢产物的细菌的总称,并且已知其通过保持肠内各种肠微生物的恒定比例并促进肠运动而具有生理功能,例如改善便秘、提高免疫力等。由于乳酸菌通常易受温度、酸度、压力等的影响,取决于流通过程或储存环境,乳酸菌的量可能达不到食品和药品安全部的推荐含量。因此,为了提高乳酸菌的稳定性,对乳酸菌的制造方法进行了持续的研究(专利文献1)。在此,在乳酸菌的培养中,由于乳酸菌的活细胞数根据培养条件而有很大的差异,因此能够提高乳酸菌的可培养性的条件的研究也在一直持续。
使用红参进行了各种研究,本发明人等确认了在乳酸菌用培养基中使用除去红参的水溶性成分的红参来源的膳食纤维时,乳酸菌的活细胞数增加、且可培养性大幅提高,从而完成了本发明。
[现有技术文献]
[专利文献]
(专利文献1)韩国专利公开第10-2048690号
发明内容
【技术问题】
本发明的目的是提供一种具有改进的乳酸菌可培养性的乳酸菌用培养基以及使用该培养基培养乳酸菌的方法。
【技术方案】
本发明的一个方面提供了一种乳酸菌用培养基,包含红参来源的膳食纤维。
本发明的另一方面提供了一种培养乳酸菌的方法,该方法包含在含有红参来源的膳食纤维的乳酸菌用培养基中培养乳酸菌。
【有益效果】
根据本发明,通过在乳酸菌用培养基中使用红参来源的膳食纤维,乳酸菌的活细胞数增加,因此可以提高可培养性。另外,根据本发明的培养乳酸菌的方法,由于在相同条件下通过使用该培养基提高了可培养性,因此在成本方面存在优势。
附图说明
图1表示实施例和比较例的乳双歧杆菌的培养结果。
图2表示实施例和比较例的短双歧杆菌的培养结果。
具体实施方式
在下文中,将更详细地描述本发明以帮助理解本发明。
基于发明人可以适当地定义术语的概念以便以最佳方式描述其自己的发明的原则,在本说明书和权利要求中使用的术语和词语不应被解释为限于典型的或字典的含义,而是应被解释为具有符合本公开的技术精神的含义和概念。
1.乳酸菌用培养基
本发明的一个方面提供了一种乳酸菌用培养基,包含红参来源的膳食纤维。
在本发明中,“红参”是指蒸制的人参(Panaxginseng)。具体地,红参可以通过用蒸汽蒸人参的根,然后干燥蒸过的根来制备,并且通过本领域使用的常规制备方法使用人参作为原料制备的任何物质都可以用于本发明。红参当通过上述相同的方法制备时可略带红色,并且可浓缩并含有人参的成分。红参中所含有的活性成分,例如皂苷,其量和类型可以与人参中所含的活性成分不同,并且红参具有比人参更少的苦味和更强的甜味或更好的质地且更易于储存。人参可以不具限制地使用,例如栽培的或商业上可获得的人参。
在本发明中,“膳食纤维”是指不被人消化酶消化,而被排出体外的高分子碳水化合物,并且可以包含例如果胶、树胶、纤维素、木质素等。
在本发明中,“红参来源的膳食纤维”是指从红参中获得的膳食纤维,其获得方法没有限制。具体而言,红参来源的膳食纤维可通过常规方法从红参中获得和制备,并可从红参原料、红参提取物或红参提取过程中产生的残渣中获得。例如,可以从红参提取过程中的残渣中提取并获得红参来源的膳食纤维,但是当从制备用于各种目的红参提取物过程中产生的残渣中获得膳食纤维时,在更有效和经济方面是有利的。
红参来源的膳食纤维,可以使用离心机、压滤机、压带机、干燥机等,从红参提取过程中的残渣中提取得到,但并不特别限定于此。
当使用干燥机从残渣中提取红参来源的膳食纤维时,使用干燥机的干燥工序可以在80℃以上、90℃以上、或100℃至200℃下进行,但并不特别限定于此。
红参提取过程可以使用所有常规已知的提取方法进行,例如溶剂提取、超声提取、过滤、和回流提取,并且例如可以使用溶剂提取或回流提取。萃取过程可以重复几次。
红参提取过程可以使用水、有机溶剂、超临界流体、其混合物等作为提取溶剂来进行。有机溶剂可以是醇,优选C1–C4低级醇、己烷(正己烷)、醚、甘油、丙二醇、丁二醇、乙酸乙酯、乙酸甲酯、二氯甲烷、氯仿、乙酸乙酯、苯、混合溶剂等,例如,乙醇。例如,当水和有机溶剂的混合物用作提取溶剂时,水和有机溶剂的混合物可以具体地为水和C1–C4低级醇的混合物,更具体地为水和乙醇的混合物。
红参提取过程可以在0℃至100℃,具体地40℃至90℃,更具体地80℃至90℃下进行,但不限于此。红参提取过程可以进行6小时至36小时,具体地7小时至24小时,更具体地8小时至14小时,但不限于此。提取可以进行1至8次,具体地1至6次,且更具体地1至5次,但不限于此。
此外,红参来源的膳食纤维中除了含有纤维外,还可含有人参皂苷如Rb1、Rg3、Rg1、和Re,红参多糖等成分。因此,本发明的红参来源的膳食纤维可以通过人参皂苷、红参多糖等发挥各种生理活性功能,也可以通过普通纤维发挥功能。在本发明中,将红参来源的膳食纤维用作乳酸菌用培养基,因此具有良好的可培养性。
本发明的乳酸菌用培养基中可以包含0.001%(w/v)至5%(w/v)的红参来源的膳食纤维。具体地,红参来源的膳食纤维可以以0.005%(w/v)至4.5%(w/v)、0.01%(w/v)至4%、0.1%(w/v)至3.5%、或1%至3%的量被包含,但不限于此。由于本发明的乳酸菌用培养基包含上述含量范围的红参来源的膳食纤维,因此通过促进培养基中培养的乳酸菌的生长而增加活细胞数的效果非常好。特别是,即使红参来源的膳食纤维的含量在上述范围内相对低,乳酸菌增殖促进效果也优异,因此,根据培养目的,可以适当选择并调整膳食纤维的含量来使用。
在本发明中,“乳酸菌”是产生大量乳酸作为代谢物的细菌的通称,并且可以包含,例如,乳杆菌属、乳球菌属、肠球菌属、链球菌属、明串珠菌属、片球菌属、双歧杆菌属等。
作为例子,乳酸菌可以包含嗜酸乳杆菌、干酪乳杆菌、格氏乳杆菌、德氏乳杆菌保加利亚亚种、瑞士乳杆菌、发酵乳杆菌、副干酪乳杆菌、植物乳杆菌、罗伊氏乳杆菌、鼠李糖乳杆菌、唾液乳杆菌、德氏乳杆菌乳亚种、乳酸乳球菌、乳酸乳球菌奶油亚种、乳酸乳球菌乳亚种丁二酮变种、乳酸乳球菌乳亚种、屎肠球菌、粪肠球菌、嗜热链球菌、肠膜明串珠菌、假肠膜明串珠菌、两歧双歧杆菌、短双歧杆菌、长双歧杆菌、动物双歧杆菌乳亚种、婴儿双歧杆菌、乳双歧杆菌、凝结芽孢杆菌等,但不限于上述乳酸菌,而包括可用于食品的所有乳酸菌。
本发明的培养基只要是通常用于培养乳酸菌的培养基,就没有特别的限制,例如可以使用MRS、BL、M17、NB、BHI培养基等。由于本发明的培养基包含红参来源的膳食纤维和培养乳酸菌常用培养基中所含有的成分,使用该培养基培养的乳酸菌的活细胞数可以显著增加,从而促进乳酸菌的生长并提高可培养性。
本发明的培养基可以进一步包含选自由糖、氮源、微量元素和碳酸盐所组成的组的一或更多个作为培养乳酸菌所需的其它成分。
糖质成分可为乳糖、果糖、蔗糖、葡萄糖、麦芽糖、海藻糖等的混合。
氮源成分可为酵母提取物、大豆蛋白胨、酪蛋白胨等。
微量元素可包含二磷酸钾、硫酸镁、硫酸锰、氯化钙、柠檬酸钾、乙酸钠、L-抗坏血酸、L-谷氨酸、L-半胱氨酸盐酸盐、聚山梨醇酯80等。
碳酸盐可以是碳酸钾、碳酸钙、碳酸钠等。
2.乳酸菌的培养方法
本发明的另一方面提供了一种使用乳酸菌用培养基培养乳酸菌的方法。
乳酸菌的培养方法包含在含有红参来源的膳食纤维的培养基中培养乳酸菌。
对于“培养基”,由于可以应用与上述相同的内容,因此将省略详细描述。
首先,在培养基中培养乳酸菌。
接种可以通过在培养基中以107CFU/mL至1011CFU/mL、5×107CFU/mL至1010CFU/mL或108CFU/mL至109CFU/mL的浓度接种乳酸菌来进行,但不限于此,并且待接种的乳酸菌的浓度可以根据乳酸菌的培养目的、培养时间等适当地选择。
然后,培养乳酸菌。
培养步骤可以基于本领域通常用于培养乳酸菌的常规条件,并且不受特别限制。
例如,培养温度条件可以是30℃至40℃、35℃至40℃、或35℃至37℃的温度。乳酸菌可以在培养相对湿度条件下培养,相对湿度条件为65%至85%、70%至80%、或72%至78%。培养步骤可以是在厌氧条件下,例如在用氮气或二氧化碳气体代替空气的条件下,培养乳酸菌。用于培养乳酸菌的培养时间可以是6至48小时、10至40小时、或15至24小时。
通过本发明的培养方法培养乳酸菌时,促进了乳酸菌的增殖,同时具有使乳酸菌的活细胞数急遽增加的效果。即,与在以往的乳酸菌用培养基中培养乳酸菌的情况相比,在通过本发明的培养方法培养乳酸菌的情况下,具有活细胞数增加16%至59%的优点。
以下,通过实施例和比较例对本发明进行详细说明。
但是,以下的制备例和实验例仅是对本发明的说明,本发明的内容并不限于以下的制备例和实验例。
实施例1
红参加入纯净水,在87℃的温度下提取24小时以分离红参提取物和残渣。为了获得红参膳食纤维(DF),通过脱水过程从除红参提取物外的红参残渣中除去水,并使用干燥器在110℃的温度下干燥所述残渣,以获得红参膳食纤维(DF)。
本发明的培养基是通过将红参膳食纤维加入到通常用于培养乳酸菌的MRS培养基中而制备的。具体地,制备添加了浓度为总培养基的1%(w/v)的红参膳食纤维的培养基(实施例1-1)、添加了浓度为2%(w/v)的红参膳食纤维的培养基(实施例1-2)、和添加了浓度为3%(w/v)的红参膳食纤维的培养基(实施例1-3),然后用于乳酸菌的培养实验。
比较例1和2
比较例1和2的培养基是通过在常规MRS培养基中加入来自红参的其它成分而不添加实施例1的红参来源的膳食纤维而制备的。具体地,比较例1-1至1-3的培养基是通过将通过收集在人参蒸制过程中产生的蒸制人参流出物并随后对其灭菌而获得的“红参来源液(CW)”以分别为1、5和10%(w/v)的浓度加入至MRS培养基而制备。比较例2-1至2-3的培养基是通过将通过离心蒸煮过的人参流出物以除去淀粉而浓缩的“红参来源浓缩物(CCW)”以分别为1、2和3%(w/v)的浓度加入至MRS培养基而制备。
实验例
在实施例和比较例的培养基中接种并培养乳酸菌,在MRS琼脂平板上测定活细胞数。使用乳双歧杆菌IDCC4301菌株(LA4301)和短双歧杆菌IDCC 4401菌株(BR 4401)作为乳酸菌。将各乳酸菌株培养至5×109CFU/ml,以5%(v/v)接种于实施例和比较例的培养基中,在37℃、75%相对湿度、厌氧条件下培养40小时,测定活细胞数的变化。在对照组中,使用未处理组不添加红参膳食纤维测量活细胞数的变化,并以百分比计算与对照组相比活细胞数的变化率。结果示于下表1中。
[表1]
结果,在含有红参来源液或红参来源浓缩物的比较例1-1至1-3和比较例2-1至2-3的培养基中,乳酸菌的活细胞数与对照组相比没有显著增加或减少。即,在培养乳双歧杆菌时,比较例中红参来源液或红参来源浓缩物的含量越增加,活细胞数越有增加的倾向,但其值不大。在含有3%(w/v)红参来源浓缩液的比较例2-3培养基中,活细胞数减少15%,促进乳酸菌培养的效果没有显著地表现出来。另外,确认了在培养短双歧杆菌时,在比较例的培养基中,所有的活细胞数减少,不适合于乳酸菌的培养。
另一方面,在含有本发明的红参膳食纤维的实施例1-1至1-3的培养基中,无论是培养乳双歧杆菌的情况还是培养短双歧杆菌的情况,乳酸菌的活细胞数都大幅度增加。即,测得活细胞数的增加率为16%以上59%以下,且在本发明的培养基中培养乳酸菌时,确认了其可培养性大幅提高。另外,即使使用含有1%(w/v)的少量红参膳食纤维的培养基(实施例1-1),也测得活细胞数的增加率高。因此,根据培养目的,可以以各种方式设定合适的培养条件,即使使用少量的红参膳食纤维,也可以获得优异的效果,因此可以预期,即使在成本方面也具有优势。
Claims (10)
1.一种乳酸菌用培养基,包括红参来源的膳食纤维。
2.如权利要求1所述的乳酸菌用培养基,其中所述乳酸细菌包括选自嗜酸乳杆菌、干酪乳杆菌、格氏乳杆菌、德氏乳杆菌保加利亚亚种、瑞士乳杆菌、发酵乳杆菌、副干酪乳杆菌、植物乳杆菌、罗伊氏乳杆菌、鼠李糖乳杆菌、唾液乳杆菌、德氏乳杆菌乳亚种、乳酸乳球菌、乳酸乳球菌奶油亚种、乳酸乳球菌乳亚种丁二酮变种、乳酸乳球菌乳亚种、屎肠球菌、粪肠球菌、嗜热链球菌、肠膜明串珠菌、假肠膜明串珠菌、两歧双歧杆菌、短双歧杆菌、长双歧杆菌、动物双歧杆菌乳亚种、婴儿双歧杆菌、乳双歧杆菌、和凝结芽孢杆菌,所组成的组的至少一个。
3.如权利要求1所述的乳酸菌用培养基,其中所述红参来源的膳食纤维是通过在红参提取过程中从残渣中提取膳食纤维而获得的。
4.如权利要求3所述的乳酸菌用培养基,其中所述红参来源的膳食纤维,是使用选自由离心机、压滤机、压带机和干燥机所组成的组的至少一个,从红参提取过程中的残渣中提取。
5.一种培养乳酸菌的方法,包括在含有红参来源的膳食纤维的乳酸菌用培养基中接种和培养乳酸菌。
6.如权利要求5所述的培养乳酸菌的方法,其中所述乳酸细菌包括选自嗜酸乳杆菌、干酪乳杆菌、格氏乳杆菌、德氏乳杆菌保加利亚亚种、瑞士乳杆菌、发酵乳杆菌、副干酪乳杆菌、植物乳杆菌、罗伊氏乳杆菌、鼠李糖乳杆菌、唾液乳杆菌、德氏乳杆菌乳亚种、乳酸乳球菌、乳酸乳球菌奶油亚种、乳酸乳球菌乳亚种丁二酮变种、乳酸乳球菌乳亚种、屎肠球菌、粪肠球菌、嗜热链球菌、肠膜明串珠菌、假肠膜明串珠菌、两歧双歧杆菌、短双歧杆菌、长双歧杆菌、动物双歧杆菌乳亚种、婴儿双歧杆菌、乳双歧杆菌、和凝结芽孢杆菌,所组成的组的至少一个。
7.如权利要求5所述的培养乳酸菌的方法,其中所述红参来源的膳食纤维是通过在红参提取过程中从残渣中提取膳食纤维而获得的。
8.如权利要求5所述的培养乳酸菌的方法,其中,所述培养在35℃至40℃的温度下进行。
9.如权利要求5所述的培养乳酸菌的方法,其中,所述培养在厌氧条件下进行。
10.如权利要求5所述的培养乳酸菌的方法,其中,所述培养进行6至48小时。
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