CN115216411A - 含有阿洛酮糖的细菌、真菌和酵母生长抑制剂 - Google Patents

含有阿洛酮糖的细菌、真菌和酵母生长抑制剂 Download PDF

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CN115216411A
CN115216411A CN202211012604.9A CN202211012604A CN115216411A CN 115216411 A CN115216411 A CN 115216411A CN 202211012604 A CN202211012604 A CN 202211012604A CN 115216411 A CN115216411 A CN 115216411A
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aspergillus
fermented alcoholic
alcoholic beverage
saccharide
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崔钟珉
金秀庭
朴允卿
朴政圭
边成倍
沈东锡
李仁�
朴承源
郑东澈
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CJ CheilJedang Corp
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Abstract

本申请涉及包含含有阿洛酮糖的糖类的微生物生长抑制剂,以及含有该生长抑制剂的发酵饮料。

Description

含有阿洛酮糖的细菌、真菌和酵母生长抑制剂
本专利申请是申请号为2017800619403、申请日为2017年9月28日、发明名称为“含有阿洛酮糖的细菌、真菌和酵母生长抑制剂”的专利申请的分案申请。
技术领域
本发明涉及用于细菌、真菌和酵母的生长抑制剂,该生长抑制剂包含含有阿洛酮糖的糖类。
背景技术
发酵食品富含活的微生物并持续进行发酵。随着时间的推移,发酵食品中微生物的数量发生了显著的变化,从而引起品质变化,如酒精含量或味道的变化。因此,使发酵食品的流通(distribution)和销售受到限制。
具体而言,未加工的马格利酒(makgeolli)(其为发酵饮料)的保质期很短,为7至10天,从而使其难以保持长期销售。此外,葡萄酒(wine)(一种发酵的酒精饮料)由于后发酵而不能保持品质不变,因此需要灭菌过程或低温储存设备以防止此类品质变化引起感官性能劣化或成本增加。
本发明人致力于抑制发酵的酒精饮料的后发酵。作为结果,本发明人发现添加阿洛酮糖可以抑制用于马格利酒的发酵的真菌(例如,米曲霉(Aspergillus oryzae)、泡盛曲霉(Asp.awamori)、紫红曲霉(Monascus purpureus)、红色红曲霉(Monascus ruber)和米根霉(Rhizopus oryzae))、细菌(例如,干酪乳杆菌(Lactobacillus casei)和乳酸乳球菌乳亚种(Lactococcus lactis subsp.lactis))和酵母(例如,酿酒酵母(Saccharomycescerevisiae))的生长,并且可以抑制用于葡萄酒的发酵的酵母(例如,酿酒酵母(Saccharomyces cerevisiae)和巴斯德酵母(Saccharomyces pastorianus))的生长,从而抑制发酵食品(例如,发酵的酒精饮料,如马格利酒和葡萄酒)的后发酵,并由此完成了本发明。
[相关文献]
[专利文献]
韩国专利No.10-1352025
发明内容
技术问题
本发明的一个目的是提供一种用于微生物的生长抑制剂,该生长抑制剂包含含有阿洛酮糖的糖类。
本申请的另一个目的是提供包含该生长抑制剂的发酵的酒精饮料。
本申请的又一个目的是提供用于抑制发酵的酒精饮料的后发酵的方法,该方法包括将该生长抑制剂添加到发酵的酒精饮料中。
技术方案
根据本发明的一个方面,用于微生物的生长抑制剂包含含有阿洛酮糖的糖类,其中所述微生物包括选自由下列细菌、真菌和酵母组成的组中的至少一者:
(i)作为细菌的干酪乳杆菌和乳酸乳球菌乳亚种;
(ii)作为真菌的米曲霉、泡盛曲霉、紫红曲霉、红色红曲霉和米根霉;以及
(iii)作为酵母的酿酒酵母和巴斯德酵母。
本文所用的阿洛酮糖可以是直接从天然产物中提取的,或者可以是化学或生物合成的,但不限于此。此外,阿洛酮糖可以是以晶体形式或含有阿洛酮糖的糖浆形式(即,以液体形式)提供的。
相对于100重量份的基于干固体(DS)含量的糖类,阿洛酮糖的含量可以为50重量份至100重量份。具体而言,相对于100重量份的基于干固体含量的糖类,阿洛酮糖的含量可以为70重量份至100重量份、90重量份至100重量份、95重量份至100重量份、98重量份至100重量份、98.5重量份至100重量份、99重量份至100重量份或99.5重量份至100重量份。
除了阿洛酮糖外,糖类还可包含至少一种甜味剂。甜味剂的实例可包括任意已知的甜味剂(例如,单糖、二糖、低聚糖、糖醇和高强度甜味剂),但不限于此。单糖的实例可包括阿拉伯糖、木糖、果糖、塔格糖、阿洛糖和半乳糖。二糖是指由连接在一起的两个单糖单元组成的一组碳水化合物,并且其实例可包括乳糖、麦芽糖、海藻糖、松二糖和纤维二糖。低聚糖是指由连接在一起的3个或更多个单糖单元组成的一组碳水化合物,并且其实例可包括低聚果糖、低聚异麦芽糖、低聚木糖、低聚龙胆糖、低聚麦芽糖和低聚半乳糖。糖醇是指通过还原糖类中的羰基而得到的化合物,并且其实例可包括赤藓糖醇、木糖醇、阿拉伯糖醇、甘露糖醇、山梨糖醇、麦芽糖醇和乳糖醇。高强度甜味剂是指甜度为蔗糖的十倍以上的甜味剂,并且其实例可包括阿斯巴甜、安赛蜜、莱苞迪甙A和三氯蔗糖,但不限于此。在另一个实施方案中,根据本发明的糖类可以不含蔗糖、葡萄糖或它们的组合。
生长抑制剂可以抑制干酪乳杆菌或乳酸乳球菌乳亚种的生长,使得在37℃下在MRS培养基中培养12小时、24小时或48小时后测量的单个细胞集落数目为培养0小时的单个细胞集落初始数目的300%以下、200%以下、180%以下、150%以下、130%以下、120%以下或100%以下。
生长抑制剂可以抑制米曲霉、泡盛曲霉、紫红曲霉、红色红曲霉或米根霉的生长,使得在25℃下在PD培养基中培养24小时、48小时或72小时后测量的单个细胞集落数目为培养0小时的单个细胞集落初始数目的3,500%以下、2,000%以下、500%以下、400%以下、300%以下、200%以下、180%以下、150%以下、130%以下、120%以下、110%以下或100%以下。
生长抑制剂可以抑制酿酒酵母或巴斯德酵母的生长,使得在25℃下在YM培养基中培养24小时、48小时或72小时后测量的单个细胞集落数目为培养0小时的单个细胞集落初始数目的400%以下、350%以下、300%以下、200%以下、150%以下、130%以下、120%以下、110%以下或100%以下。
根据本发明的另一个方面,发酵的酒精饮料包含根据本发明的生长抑制剂。
发酵的酒精饮料可包括通过将产生醇的微生物接种到水果或谷物中,然后在特定条件下发酵而得到的任意已知的发酵的酒精饮料。具体而言,根据本发明的发酵的酒精饮料可以为马格利酒、咚咚酒(dongdongju)、低度米酒(takju)、葡萄酒或啤酒,更具体而言为马格利酒或葡萄酒。
发酵的酒精饮料可以为这样的酒精饮料,其由选自由下列细菌、真菌和酵母组成的组中的至少一种微生物发酵:(i)作为细菌的干酪乳杆菌和乳酸乳球菌乳亚种;(ii)作为真菌的米曲霉、泡盛曲霉、紫红曲霉、红色红曲霉和米根霉;以及(iii)作为酵母的酿酒酵母和巴斯德酵母。
发酵的酒精饮料可以为这样的发酵的酒精饮料,其包含选自由下列细菌、真菌和酵母组成的组中的至少一种微生物:(i)作为细菌的干酪乳杆菌和乳酸乳球菌乳亚种;(ii)作为真菌的米曲霉、泡盛曲霉、紫红曲霉、红色红曲霉和米根霉;以及(iii)作为酵母的酿酒酵母和巴斯德酵母。
发酵的酒精饮料还可包含通常用于发酵的酒精饮料的食品成分。具体而言,根据本发明的发酵的酒精饮料还可包含选自由下列组成的组中的至少一种食品成分:纯净水、低聚糖、大米、淀粉(例如麦芽糖糊精)、二氧化碳气体、阿斯巴甜、有机酸(例如柠檬酸)和植物提取物。
在发酵的酒精饮料中,生长抑制剂、阿洛酮糖、糖类、细菌、真菌和酵母与上述相同。
根据本发明的又一个方面,用于抑制发酵的酒精饮料的后发酵的方法包括将根据本发明的生长抑制剂添加到发酵的酒精饮料中。
现在,将通过制备发酵的酒精饮料的方法来更详细地描述本发明的实施方案。如上所述,发酵的酒精饮料的实例包括代表性的马格利酒和葡萄酒。首先,将通过制造马格利酒的方法来更详细地描述本发明的实施方案,所述方法包括以下步骤:
(1)通过使用细菌、酵母或真菌发酵未熟或熟的淀粉原料来制备母液;
(2)通过添加淀粉原料、糖化剂和水来使步骤(1)中制备的母液糖化;
(3)使步骤(2)中得到的糖化产物发酵,然后进行熟化;
(4)通过将步骤(3)中得到的熟化产物过筛来除去未溶解的淀粉固体;以及
(5)通过向步骤(4)中得到的过筛产物中添加阿洛酮糖、糖类和水来制备最终产物。
如在上述制备马格利酒的方法中,由于根据本发明的抑制除马格利酒之外的发酵的酒精饮料(如葡萄酒)的后发酵的方法基本上与典型的葡萄酒制造方法相同,不同之处在于添加了根据本发明的糖类和阿洛酮糖而不是典型的糖类或甜味剂,因此将省略对其描述。
在用于抑制发酵的酒精饮料的后发酵的方法中,生长抑制剂、阿洛酮糖、糖类、细菌、真菌和酵母与上述相同。
有益的效果
根据本发明,可以抑制发酵的酒精饮料中微生物的生长,从而抑制发酵的酒精饮料的后发酵。具体而言,当使用阿洛酮糖代替通常添加到发酵的酒精饮料(如马格利酒或葡萄酒)中的糖类(如蔗糖)时,本发明可以得到这样的效果:防止在发酵的酒精饮料的储存或流通过程中可能发生的无意的品质劣化,以及消除对用于提高储存稳定性的单独工艺或设备的需要,从而在降低制造成本的同时简化制造工艺。
附图说明
图1和图2为示出如在使用葡萄糖或蔗糖作为糖类的比较例和使用阿洛酮糖作为糖类的实施例的样品上所测量的细菌目标物(干酪乳杆菌(图1)和乳酸乳球菌乳亚种(图2))的时间依赖性CFU值的图。
图3至图7为示出如在使用葡萄糖或蔗糖作为糖类的比较例和使用阿洛酮糖作为糖类的实施例的样品上所测量的真菌目标物(米曲霉(图3)、泡盛曲霉(图4)、紫红曲霉(图5)、红色红曲霉(图6)和米根霉(图7))的时间依赖性CFU值的图。
图8和图9为示出如在使用葡萄糖或蔗糖作为糖类的比较例和使用阿洛酮糖作为糖类的实施例的样品上所测量的酵母目标物(酿酒酵母(图8)和巴斯德酵母(图9))的时间依赖性CFU值的图。
具体实施方式
在下文中,将参照例子更详细地描述本发明。然而,应当注意,提供这些例子仅用于说明,而不应以任何方式解释为限制本发明。此外,提供这些例子是为了使本领域普通技术人员更完整地理解本发明。
例子
实验例1:用于发酵酒精饮料的发酵的微生物和培养基的准备
为了评价对于发酵的酒精饮料的后发酵的抑制,确定了对发酵的酒精饮料中常用的微生物的生长的抑制程度。
具体而言,通过典型的方法在培养基(表1)中将得自韩国微生物培养中心(KCCM)的各微生物(细菌、真菌和酵母)传代培养三次或四次以增强活性。然后,将培养的微生物接种到培养基(表1)中,然后进行过度培养至浓度为105CFU/mL以上,从而得到菌株母液(表1)。
表1
Figure BDA0003811497650000061
实验例2:根据糖类组成确认微生物的生长
为了比较仅根据添加的糖类的种类而非由于发酵的酒精饮料所含的酒精和其他添加剂而产生的偏差所致的对于微生物生长的抑制程度,仅改变添加至各微生物培养基中的糖类的种类并进行研究。
具体而言,作为适合于各菌株的培养基的糖类,将含水结晶葡萄糖(纯度98重量%以上,CJ Cheiljedang)、蔗糖(纯度98重量%以上,CJ Cheiljedang)和结晶阿洛酮糖(纯度98重量%以上,CJ Cheiljedang)添加到各培养基中,从而制备改性培养基(表2至表4)。然后,向100mL所制备的改性培养基中接种1mL实验例1中制备的各菌株的母液,然后观察各菌株的细胞数目的时间依赖性变化。也就是说,以相同的方式在表2中列出的MRS培养基、表3中列出的PDB培养基和表4中列出的YMB培养基中分别接种细菌(干酪乳杆菌和乳酸乳球菌乳亚种)、真菌(米曲霉、泡盛曲霉、紫红曲霉、红色红曲霉和米根霉)和酵母(酿酒酵母和巴斯德酵母)。然后,将接种有细菌的各培养基在37℃储存12小时、24小时和48小时,以用作测量单个细胞集落数目的样品,而将接种有真菌或酵母的各培养基在25℃储存24小时、48小时和72小时,以用作测量单个细胞集落数目的样品。
通过典型的微生物试验进行单个细胞集落数目的测量。具体而言,通过10倍稀释法使用9mL 0.9%的无菌生理盐水逐渐稀释1mL的各样品,从而得到稀释的样品。然后,将1mL稀释的样品置于培养皿中,并将25mL添加有琼脂的标准培养基倒入培养皿中,以便充分混合并固化,从而得到接种的样品。这里,MRSA、PDA和YMA作为添加有琼脂的标准培养基分别用于细菌、真菌和酵母。
此后,将测量的细胞集落数目乘以稀释因子,从而计算每mL样品的集落形成单位(CFU)的值。
表2
Figure BDA0003811497650000081
表3
Figure BDA0003811497650000082
表4
Figure BDA0003811497650000083
作为结果,证实了使用阿洛酮糖作为糖类的实施例1至3的9种菌株的细胞集落数目显著低于比较例的菌株的细胞集落数目。此外,证实了在典型培养基中使用葡萄糖作为糖类的比较例1-1、2-1和3-1的菌株表现出微生物的高度生长,并且使用蔗糖作为糖类的比较例1-2、2-2和3-2的菌株表现出与使用葡萄糖作为糖类的接种的菌株无显著差异的微生物生长。
具体而言,在使用葡萄糖作为糖类的培养基中(比较例1-1)和使用蔗糖作为糖类的培养基中(比较例1-2),作为细菌的干酪乳杆菌和乳酸乳球菌乳亚种的细胞数增加了40.6倍至137倍。与之相比,在使用阿洛酮糖作为糖类的培养基中,细菌的细胞数仅增加了1.3倍至2.2倍。因此,基本上没有观察到菌株生长(图1和图2)。
此外,在使用葡萄糖作为糖类的培养基中(比较例2-1)和使用蔗糖作为糖类的培养基中(比较例2-2),作为真菌的米曲霉、泡盛曲霉、紫红曲霉、红色红曲霉和米根霉的细胞数分别增加了9.8倍至148.5倍和5.9倍至81.1倍。与之相比,在使用阿洛酮糖作为糖类的培养基中,真菌的细胞数仅增加了1.6倍至9.8倍(实施例2)。因此,观察到了统计学上显著的微生物生长的抑制(图3至图7)。
最后,在使用葡萄糖作为糖类的培养基中(比较例3-1)和使用蔗糖作为糖类的培养基中(比较例3-2),作为酵母的酿酒酵母和巴斯德酵母的细胞数增加了27.4倍至134.8倍。与之相比,在使用阿洛酮糖作为糖类的培养基中,酵母的细胞数仅增加了1.3倍至3倍。因此,基本上没有观察到菌株生长(图8和图9)。
对于各菌株,在各培养基中培养0小时、12小时、24小时、48小时和72小时后测量单个菌落的数目。结果示于表5中。
表5
Figure BDA0003811497650000091
Figure BDA0003811497650000101
总之,可以看出,当使用阿洛酮糖代替通常用作添加到诸如马格利酒之类的发酵的酒精饮料中的甜味剂的蔗糖时,可以降低在发酵的酒精饮料的制造或流通过程中可能发生的微生物生长所引起的后发酵。
虽然本文已经描述了一些示例性实施方案,但是本领域技术人员应当理解,这些实施方案仅以说明的方式给出,并且在不脱离本发明的精神和范围的情况下可以进行各种修改、变化和改变。因此,实施方案和附图不应被理解为对本发明的技术精神的限制,而应被理解为对本发明的技术精神的说明。本发明的范围应根据以下所附权利要求来解释,以涵盖来源于所附权利要求及其等同物的所有修改或变化。

Claims (8)

1.一种用于微生物的生长抑制剂,包含:
含有阿洛酮糖的糖类,
其中所述微生物包括选自由下列细菌、真菌和酵母组成的组中的至少一者:
(i)作为细菌的干酪乳杆菌和乳酸乳球菌乳亚种;
(ii)作为真菌的米曲霉、泡盛曲霉、紫红曲霉、红色红曲霉和米根霉;以及
(iii)作为酵母的酿酒酵母和巴斯德酵母。
2.根据权利要求1所述的生长抑制剂,其中相对于100重量份的基于干固体含量的所述糖类,所述阿洛酮糖的含量为50重量份至100重量份。
3.根据权利要求1所述的生长抑制剂,其中所述糖类不含蔗糖、葡萄糖或它们的组合。
4.一种发酵的酒精饮料,其包含根据权利要求1至3中任一项所述的生长抑制剂。
5.根据权利要求4所述的发酵的酒精饮料,其中所述发酵的酒精饮料为马格利酒或葡萄酒。
6.根据权利要求4所述的发酵的酒精饮料,其中使用选自由下列细菌、真菌和酵母组成的组中的至少一种微生物来发酵所述发酵的酒精饮料:
(i)作为细菌的干酪乳杆菌和乳酸乳球菌乳亚种;
(ii)作为真菌的米曲霉、泡盛曲霉、紫红曲霉、红色红曲霉和米根霉;以及
(iii)作为酵母的酿酒酵母和巴斯德酵母。
7.根据权利要求4所述的发酵的酒精饮料,其中所述发酵的酒精饮料包含选自由下列细菌、真菌和酵母组成的组中的至少一种微生物:
(i)作为细菌的干酪乳杆菌和乳酸乳球菌乳亚种;
(ii)作为真菌的米曲霉、泡盛曲霉、紫红曲霉、红色红曲霉和米根霉;以及
(iii)作为酵母的酿酒酵母和巴斯德酵母。
8.一种抑制发酵的酒精饮料的后发酵的方法,包括:将根据权利要求1至3中任一项所述的生长抑制剂添加到所述发酵的酒精饮料中。
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