CN109890954A - 具有透明质酸产生促进能力的乳酸菌 - Google Patents
具有透明质酸产生促进能力的乳酸菌 Download PDFInfo
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- CN109890954A CN109890954A CN201780044028.7A CN201780044028A CN109890954A CN 109890954 A CN109890954 A CN 109890954A CN 201780044028 A CN201780044028 A CN 201780044028A CN 109890954 A CN109890954 A CN 109890954A
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- China
- Prior art keywords
- lactic acid
- acid bacteria
- hyaluronic acid
- lactobacillus
- promotion ability
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- NPFOYSMITVOQOS-UHFFFAOYSA-K iron(III) citrate Chemical compound [Fe+3].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NPFOYSMITVOQOS-UHFFFAOYSA-K 0.000 description 1
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- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
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- 238000004519 manufacturing process Methods 0.000 description 1
- 239000008268 mayonnaise Substances 0.000 description 1
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- HOVAGTYPODGVJG-ZFYZTMLRSA-N methyl alpha-D-glucopyranoside Chemical compound CO[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O HOVAGTYPODGVJG-ZFYZTMLRSA-N 0.000 description 1
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- 229940120668 salicin Drugs 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
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- 238000010186 staining Methods 0.000 description 1
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- 239000005720 sucrose Substances 0.000 description 1
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- 229940034610 toothpaste Drugs 0.000 description 1
- 239000000606 toothpaste Substances 0.000 description 1
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
Abstract
本发明的目的在于，提供具有高的透明质酸产生促进能力、可以合适地用于饮食品的属于乳杆菌属的乳酸菌及含有其处理物的饮食品。着眼于对于肌肤功能的改善而言透明质酸必不可缺这一点，对多种乳酸菌进行了筛选，结果发现存在具有高的透明质酸产生促进能力的乳酸菌，从而完成了本发明。本发明提供具有高的透明质酸产生促进能力的属于乳杆菌属的乳酸菌，更具体而言，提供格氏乳杆菌N320株(NITE BP‑02287)。另外，提供含有该乳酸菌的饮食品或以该乳酸菌作为有效成分的透明质酸产生促进剂。
Description
技术领域
本发明涉及具有高的透明质酸产生促进能力的来自于婴儿肠内的乳酸菌的菌株及含有该乳酸菌的处理物的饮料、食品。
背景技术
近年来，广泛地进行了有关防止皮肤老化的研究。对于皮肤老化的原因，年龄增长是重要的因素，然而除此以外，作为与皮肤老化相关的直接的因素还可以举出由干燥、氧化、紫外线等造成的影响。作为皮肤老化的具体现象，已知有以透明质酸为首的粘多糖类的减少、胶原蛋白的交联反应、由紫外线造成的细胞的损伤等。
此处，透明质酸广泛地存在于皮肤、关节、韧带、肺、肾脏、脑等脏器、组织中，由于保湿性高，因此是保持皮肤湿润的重要的因素(非专利文献1)。另外，已知在皮肤中存在有占整个身体的50％的量的透明质酸(非专利文献2)。
现有技术文献
非专利文献
非专利文献1：Papakonstantinou E.，：Hyaluronic acid：A Keymoleculein skinaging，Dermatoendocrinol，4(3)，253-8，2012
非专利文献2：T.C.Laurent and J.R.Fraser，“Hyaluronan，”FASEB J，Vol.6，pp.2397-2402，1992
发明内容
发明所要解决的问题
本发明人等着眼于透明质酸对于肌肤功能的改善必不可少这一点，对多种乳酸菌进行了筛选，结果发现，存在具有高的透明质酸产生促进能力的乳酸菌，从而完成了本发明。
即，本申请第一发明是具有透明质酸产生促进能力的属于乳杆菌属的乳酸菌，本申请第二发明是，本申请第一发明中记载的乳酸菌为格氏乳杆菌N320株(NITE BP-02287)。
此外，本申请的申请人还意欲提供包含该乳酸菌的饮食品。即，本申请第三发明是含有本申请第二发明中记载的乳酸菌的饮食品。另外，本申请的申请人还意欲提供以该乳酸菌作为有效成分的透明质酸产生促进剂。即，本申请第四发明是以本申请第二发明中记载的乳酸菌作为有效成分的透明质酸产生促进剂。
发明效果
本发明的乳酸菌具有透明质酸产生促进能力。
附图说明
图1是比较本发明的菌株与比较菌株的透明质酸产生促进能力的图。
图2是比较本发明的菌株与比较菌株的脱脂乳培养基增殖性的图。
具体实施方式
以下，对本发明进行详细说明。
1.格氏乳杆菌N320株(NITE BP-02287)
本发明的乳酸菌为格氏乳杆菌(lactobacillus gasseri)。特别是属于格氏乳杆菌的乳酸菌中的格氏乳杆菌N320株(NITE BP-02287)。本发明中所说的N320的记号是由日清食品控股株式会社独自对菌株赋予的编号，本格氏乳杆菌N320株是由本发明人首次分离出的菌株。
本发明的格氏乳杆菌N320株在下述的条件下保藏。
(1)保藏单位名：国家技术评估学会，专利微生物保藏中心；
(2)联系地址：〒292－0818千叶县木更津市上总镰足2－5－8122号室；
(3)保藏编号：NITE BP－02287；
(4)用于识别的标识：N320；
(5)原保藏日：2016年6月14日；
(6)转移至基于布达佩斯条约的保藏的移管日：2017年4月10日。
本发明的格氏乳杆菌N320株的菌学的性质如以下的表1及表2所示。本菌学的性质基于Bergey’s manual of systematic bacteriology Vol.2(1986)中记载的方法。表1表示有关本菌株的形状等，表2表示利用API50CH及APICHL(生物梅里埃(bioMerieux)制)试验了糖同化性的结果。表2中，“+”表示有发酵性，“－”表示无发酵性。
[表1]
(用MRS琼脂平板培养基进行37℃、48小时好氧培养时的目视观察时)
(用MRS琼脂平板培养基进行37℃、48小时好氧培养时的目视观察时)
| 革兰氏染色性 | 阳性 |
| 芽孢形成 | 阴性 |
| 气体生成 | 无 |
| 运动性 | 无 |
| 过氧化氢酶活性 | 阴性 |
| 脱脂乳凝固性 | 凝固 |
2.透明质酸产生试验
本发明的格氏乳杆菌N320株如后述的实验例所示，具有高的透明质酸产生促进能力。对于透明质酸产生促进能力的确认，利用以下的试验方法进行。
＜透明质酸产生促进能力评价中所用的受试体(培养液)的制备＞
透明质酸产生促进能力评价中所用的受试体(培养液)通过如下操作得到，即，将乳酸菌用表3所示的MRS培养基(Difco Lactobacilli MRS Broth)进行37℃、24小时培养，并进行95℃、20分钟加热杀菌后，使用氢氧化钠水溶液使pH为7.0。
＜透明质酸产生促进能力评价＞
对利用上述操作得到的试样的透明质酸产生促进作用进行了评价。即，向24孔板中以5.0×104细胞/孔的浓度接种正常人成人表皮角化细胞，达到汇合状态时，添加包含各试样组(最终浓度1％)的500μL的HuMedia-KG2培养液。添加后，温育72小时，然后利用ELISA法测定出透明质酸产生量。透明质酸产生量的测定单位设为ng/ml，测定中使用了Hyaluronan assay kit(生化学BIOBUSINESS公司制)。需要说明的是，使用乳酸菌增殖培养基(MRS)作为对照例(Control)。
[表2]
| 糖底物 | 结果 | 糖底物 | 结果 | ||
| 0 | Control | - | 25 | Esculin ferric citrate | + |
| 1 | 丙三醇 | - | 26 | 水杨苷 | + |
| 2 | 赤藓醇 | - | 27 | D-纤维二糖 | + |
| 3 | D-阿拉伯糖 | - | 28 | D-麦芽糖 | + |
| 4 | L-阿拉伯糖 | - | 29 | D-乳糖 | + |
| 5 | D-核糖 | - | 30 | D-蜜二糖 | - |
| 6 | D-木糖 | - | 31 | D-蔗糖 | + |
| 7 | L-木糖 | - | 32 | D-海藻糖 | + |
| 8 | D-阿东糖醇 | - | 33 | 菊粉 | - |
| 9 | 甲基-βD-吡喃木糖苷 | - | 34 | D-松三糖 | - |
| 10 | D-半乳糖 | + | 35 | D-棉子糖 | - |
| 11 | D-葡萄糖 | + | 36 | 淀粉 | + |
| 12 | D-果糖 | + | 37 | 糖原 | - |
| 13 | D-甘露糖 | + | 38 | 木糖醇 | - |
| 14 | L-山梨糖 | - | 39 | 龙胆二糖 | + |
| 15 | L-鼠李糖 | - | 40 | D-松二糖 | - |
| 16 | 卫矛醇 | - | 41 | D-来苏糖 | - |
| 17 | 肌醇 | - | 42 | D-塔格糖 | + |
| 18 | D-甘露醇 | - | 43 | D-岩藻糖 | - |
| 19 | D-山梨醇 | - | 44 | L-岩藻糖 | - |
| 20 | 甲基-αD-吡喃甘露糖苷 | - | 45 | D-阿糖醇 | - |
| 21 | 甲基-αD-吡喃葡萄糖苷 | - | 46 | L-阿糖醇 | - |
| 22 | N-乙酰基-葡糖胺 | + | 47 | 葡糖酸 | - |
| 23 | 扁桃苷 | + | 48 | 2-酮-葡糖酸 | - |
| 24 | 熊果苷 | + | 49 | 5-酮-葡糖酸 | - |
[表3]
| Proteose Pepton No.3 | 10g |
| 牛肉浸膏 | 10g |
| 酵母浸膏 | 5g |
| D-葡萄糖 | 20g |
| 聚山梨醇酯80 | 1g |
| 柠檬酸铵 | 2g |
| 乙酸钠 | 5g |
| 硫酸锰 | 0.1g |
| 硫酸镁 | 0.05g |
| 磷酸氢二钾 | 2g |
| 蒸馏水 | 1000ml |
3.利用脱脂乳培养基的增殖性试验
<脱脂乳培养基增殖性试验>
将前培养液接种于10％SM(脱脂乳)培养基中，对其进行37℃、24小时培养后，根据乳酸菌数评价了利用乳培养基的增殖性。
4.饮食品
可以使饮食品中含有本发明的乳酸菌而使用。本发明的乳酸菌可以特别合适地用于乳制品，例如可以考虑加入乳酸菌的发酵乳及加入乳酸菌的乳酸菌饮料。现行的关于乳及乳制品的成分标准等的部令中，作为成分标准没有特别规定乳酸菌数，然而如果是发酵乳(脱脂乳固体成分为8.0％以上的)、乳酸菌饮料(脱脂乳固体成分为3.0％以上的)，则优选为1.0×107cfu/ml以上，如果是乳酸菌饮料(脱脂乳固体成分小于3.0％的)，则优选为1.0×106cfu/ml以上，使之在乳等的发酵液中增殖、或以最终制品的形态增殖，由此可以实现上述的菌数。另外，除了可以用于加入乳酸菌的发酵乳及加入乳酸菌的乳酸菌饮料以外，还可以用于黄油等乳制品、蛋黄酱等蛋加工品、黄油蛋糕等糕点面包类等。另外，还可以合适地用于方便面、曲奇饼等加工食品中。除了上述以外，本发明的食品也可以是与所述乳酸菌一起根据需要添加适当的载体及添加剂而制剂化了的形态(例如粉末、颗粒、胶囊、片剂等)。
除了可以使一般的饮料、食品中含有本发明的乳酸菌以外，还可以使特定保健用食品、营养辅助食品等中含有本发明的乳酸菌。
另外，本发明的乳酸菌除了应用于食品以外，还可以应用于化妆水等化妆品领域、整肠剂等医药品领域、牙膏等日用品领域、青贮饲料、动物用饵食、植物液体肥料等动物饲料、植物肥料领域。
产业上的可利用性
本发明的乳酸菌(格氏乳杆菌N320株)具有高的透明质酸产生促进能力。
[实施例]
以下，给出本发明的实施例，然而本发明并不限定于以下的实施例。
＜试验例1＞透明质酸产生促进能力评价
对本发明的格氏乳杆菌N320株和本公司拥有的3种格氏乳杆菌比较菌株(N219、N220、N313)实施了透明质酸产生促进能力评价。
依照下面的步骤进行了透明质酸产生促进能力评价。对于本发明的菌株和比较菌株，分别用表3所示的MRS培养基(Difco Lactobacilli MRS Broth)进行37℃、24小时培养并进行95℃、20分钟加热杀菌后，使用氢氧化钠水溶液使pH为7.0，由此得到受试体(培养液)。
另行向24孔板中以5.0×104细胞/孔的浓度接种正常人成人表皮角化细胞，达到汇合状态时，添加包含各试样组(最终浓度1％)的500μL的HuMedia-KG2培养液。添加后，进行72小时温育，然后利用ELISA法测定出透明质酸产生量。需要说明的是，使用乳酸菌增殖培养基(MRS)作为对照例(Control)。测定中使用了Hyaluronan assay kit(生化学BIOBUSINESS公司制)。测定值的单位表示透明质酸产生量(ng/ml)。
表4及图1中表示出将乳酸菌增殖培养基(MRS培养基)设为0的各菌株的透明质酸产生量。
[表4]
| 菌株名 | 透明质酸产生量(ng/ml) |
| Control(未添加乳酸菌) | 0 |
| 格氏乳杆菌N219 | 163.3 |
| 格氏乳杆菌N220 | 7.6 |
| 格氏乳杆菌N313 | 106.8 |
| 格氏乳杆菌N320 | 332.6 |
从表4及图1可以清楚地看到，本发明的乳酸菌(格氏乳杆菌N320株)的透明质酸产生促进能力非常强，确认与其他的本公司拥有的乳酸菌相比具有更高的透明质酸产生促进能力。
＜试验例2＞脱脂乳培养基增殖性试验
对本发明的格氏乳杆菌N320株和本公司拥有的3种格氏乳杆菌比较菌株(N219、N220、N313)，实施了脱脂乳培养基增殖性试验。
向10％SM(脱脂乳)培养基中植入各菌株，对其进行37℃、24小时培养后，根据乳酸菌数评价了利用脱脂乳培养基的增殖性。
将增殖性的评价结果表示于表5及图2中。
[表5]
| 菌株名 | 菌数(×10<sup>7</sup>cfu/ml) |
| 格氏乳杆菌N219 | 0.9 |
| 格氏乳杆菌N220 | 0.9 |
| 格氏乳杆菌N313 | 15.5 |
| 格氏乳杆菌N320 | 22 |
本发明的菌株(格氏乳杆菌N320株)的菌数为22×107cfu/ml，与其他的比较菌株相比增殖性高，是在生产加入乳酸菌的发酵乳及加入乳酸菌的乳酸菌饮料方面没有问题的水平。
PCT/RO/134表
Claims (4)
1.一种乳酸菌，其具有透明质酸产生促进能力，且属于乳杆菌属。
2.根据权利要求1所述的乳酸菌，其中，
所述属于乳杆菌属的乳酸菌为格氏乳杆菌N320株，即NITE BP-02287。
3.一种饮食品，其含有权利要求2所述的乳酸菌。
4.一种透明质酸产生促进剂，其以权利要求2所述的乳酸菌作为有效成分。
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| CN102575223A (zh) * | 2009-09-02 | 2012-07-11 | 株式会社明治 | 乳杆菌属乳酸菌的增殖促进剂和/或存活性提高剂 |
| KR20140128675A (ko) * | 2013-04-29 | 2014-11-06 | 주식회사한국야쿠르트 | 피부 주름 개선 및 보습 효능을 갖는 신규의 락토바실러스 가세리 ｈｙ７０２５ 유산균 및 이를 유효성분으로 함유하는 제품 |
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| JP2010143885A (ja) * | 2008-12-22 | 2010-07-01 | Asahi Breweries Ltd | 乳酸菌およびそれらを用いた飲食物又は化粧品 |
| CN102575223A (zh) * | 2009-09-02 | 2012-07-11 | 株式会社明治 | 乳杆菌属乳酸菌的增殖促进剂和/或存活性提高剂 |
| KR20140128675A (ko) * | 2013-04-29 | 2014-11-06 | 주식회사한국야쿠르트 | 피부 주름 개선 및 보습 효능을 갖는 신규의 락토바실러스 가세리 ｈｙ７０２５ 유산균 및 이를 유효성분으로 함유하는 제품 |
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