CN1387564A - 产生抗高血压二和三肽的瑞士乳杆菌 - Google Patents
产生抗高血压二和三肽的瑞士乳杆菌 Download PDFInfo
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- CN1387564A CN1387564A CN00815393A CN00815393A CN1387564A CN 1387564 A CN1387564 A CN 1387564A CN 00815393 A CN00815393 A CN 00815393A CN 00815393 A CN00815393 A CN 00815393A CN 1387564 A CN1387564 A CN 1387564A
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- Prior art keywords
- lactobacterium helveticus
- bacterial strain
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- helveticus lbk
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Classifications
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- 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
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- A—HUMAN NECESSITIES
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- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1234—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
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- A23C9/1427—Milk preparations; Milk powder or milk powder preparations in which the chemical composition of the milk is modified by non-chemical treatment by dialysis, reverse osmosis or ultrafiltration by dialysis, reverse osmosis or hyperfiltration, e.g. for concentrating or desalting
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- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A—HUMAN NECESSITIES
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- 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
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- A—HUMAN NECESSITIES
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- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/12—Antihypertensives
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
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- C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
- C07K5/0808—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms, e.g. Val, Ile, Leu
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- 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
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- 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
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- 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
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- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
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- Food Science & Technology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
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- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- General Engineering & Computer Science (AREA)
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- Mycology (AREA)
- Biophysics (AREA)
- Water Supply & Treatment (AREA)
- Pharmacology & Pharmacy (AREA)
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- Public Health (AREA)
- Cardiology (AREA)
- Veterinary Medicine (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
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Abstract
本发明涉及一种新菌株瑞士乳杆菌LBK-16H,DSM 13137,和含有此菌株的细菌制品。该新菌株瑞士乳杆菌LBK-16H具有蛋白水解活性并能够促进一氧化氮的产生,因此可以在乳品工业中用作起始菌、并可以就此及在各种促进健康产品和药物中用作治疗性物质。
Description
发明领域
本发明涉及一种新微生物和其用途。更准确地说,本发明描述了瑞士乳杆菌(Lactobacillus helveticus)的一个新菌株、及它的生理学特性、和它在例如食品工业和制药业中的用途。
发明背景
伯杰氏系统细菌学手册(Bergey’s Manual of SystematicBacteriology),第2卷,Sneath等编,Williams和Wilkins,Baltimore,London,Los Angeles,Sydney,1984,第14部分,自1208页始,在标题“常规非产孢子革兰氏阳性杆菌”下描述了乳杆菌属(Lactobacillus)微生物的特性和分类以及菌种瑞士乳杆菌的性质。瑞士乳杆菌菌株一般从乳制品如发酵的乳制品和干酪中分离,并且它们已被常规用作生产干酪,尤其是Emmental和Gruyere型干酪,的发酵剂微生物。
现有技术中也已有瑞士乳杆菌菌株的生物学作用的描述。例如,Yamamoto等的国际专利申请WO99/16862描述了菌株瑞士乳杆菌CM4,FERM BP-6060,该菌株能够产生大量的三肽Val-Pro-Pro和/或Ile-Pro-Pro,并且具有高的细胞外蛋白酶活性。该公开文本还描述了含有上述三肽和细菌的发酵乳制品,以及通过含有这些三肽序列的所述细菌的发酵产品制备该乳制品的方法。
Nakamura等的美国专利5,449,661描述了对含有三肽序列Val-Pro-Pro的肽的制备和其在降低高血压中的用途。该肽是通过采用菌株瑞士乳杆菌JCM1004发酵脱脂乳制备的,之后该肽经层析方法纯化并被冻干。
Yamamoto等(生物化学杂志.1993,114:740)也描述了来源于微生物瑞士乳杆菌CP790的蛋白酶的纯化和性质。而且,Yamamoto等还报道了有关采用所述蛋白酶水解αs1-和β-酪蛋白及所获肽对ACE抑制作用的研究(J Dairy Sci,1994,77:917)。总共研究了25种肽,它们的分子大小和作用均有极大的差异。最有效的是从β-酪蛋白获得的三种肽,它们分别含有8、18和27个氢基酸。该研究还比较了采用菌株瑞士乳杆菌CP790发酵的乳和采用其缺失了蛋白酶活性的变种CP791发酵的乳的ACE活性,由此发现前者在自发高血压SHR大鼠中有效,但在普通大鼠品系中无效,而后者根本没有活性。
尽管对乳酸菌及瑞士乳杆菌种已有广泛研究,并且它们已被推荐用作常规的发酵剂和促进健康的物质,但在该领域中仍需要持续寻找可以在制酪业和其它食品工业中用作发酵剂和益生菌、以及可以作为天然产品使用和用于制药业的新的有效的微生物。
发明描述
本发明的目标是提供具有优秀的蛋白水解性质和生理学性质、并由此极为适于用作发酵剂和促进健康物质的瑞士乳杆菌的一个新菌株。
根据本发明,该菌株被提供给消费者,就此使用或用于食品、功能性产品或药物的生产中。
因此,本发明涉及瑞士乳杆菌LBK-16H,DSM1317。
本发明还涉及含有菌株瑞士乳杆菌LBK-16H,DSM1317的细菌制品。
本发明还涉及菌株瑞士乳杆菌LBK-16H,DSM1317在食品工业或制药业中的用途。
再有,本发明还涉及含有上述菌株或通过使用上述菌株制备的可食用产品,例如食品和药物。
本发明还涉及菌株瑞士乳杆菌LBK-16H,DSM13137作为治疗性物质的用途。
本发明还涉及菌株瑞士乳杆菌LBK-16H,DSM13137在高血压治疗中的用途。
再有,本发明还涉及菌株瑞士乳杆菌LBK-16H,DSM13137在制备抗高血压产品中的用途。
而且,本发明还涉及利用菌株瑞士乳杆菌LBK-16H,DSM 13137制备抗高血压产品的方法。
本发明基于一个新菌株瑞士乳杆菌LBK-16H,该菌株于1999年11月3日保藏在保藏单位-德意志微生物保藏中心(DSM),保藏号DSM13137,它具有如下特性:
瑞士乳杆菌菌株是革兰氏阳性、杆状长细胞。生长的温度范围是约35-45℃,最适温度约37-42℃。
在37-42℃时瑞士乳杆菌LBK-16H在乳中生长良好,产生2.5-2.9%乳酸(DL)。生长的最适pH是约4.5-7。在未进行pH调节的培养中,乳的pH降低范围为3.3至3.6。
该菌株在通常用于乳酸菌属的培养基中也生长良好,并且也可以采用柠檬酸盐作为碳源。
瑞士乳杆菌LBK-16H的碳水化合物利用情况如下:
碳水化合物 瑞士乳杆菌LBK-16H
甘油 -
赤藓糖醇 -
D-阿拉伯醣 -
L-阿拉伯糖 -
核糖 -
D-木糖 -
L-木糖 -
阿东糖醇 -
β-甲基木糖苷 -
半乳糖 +
D-葡萄糖 +
D-果糖 +
D-甘露糖 +
L-山梨糖 -
鼠李糖 -
半乳糖醇 -
肌醇 -
甘露醇 -
山梨糖醇 -
α-甲基-D-甘露糖苷 -
α-甲基-D-葡萄糖苷 -
N-乙酰葡糖胺 +
七叶苷 -
纤维二糖 -
麦芽糖 -
乳糖 +
蔗糖 -
海藻糖 -
菊粉 -
松三糖 -
D-棉子糖 -
糖原 -
木糖醇 -
瑞士乳杆菌LBK-16H具有蛋白水解活性。通过OPA方法测定(该方法在以下实施例中有述并基于邻苯二醛的蛋白水解),该菌株的蛋白水解活性在0.3-0.6级。可以利用蛋白水解活性作为发酵剂和用于生物学活性物质的制备。
瑞士乳杆菌LBK-16H对低pH值有极好的耐受性。从可保存性和生理活性的观点看,这是一个重要的性质。正如实施例中所述,该菌株可极好地耐受4甚至3的pH水平4个小时,并且其可保存性尤其在乳制品中十分优秀,在pH2这样低的pH中可保存时间高达4小时。基于这些结果,也可以推测该菌株在通过胃后可存活于消化道中,并且在结肠中仍可存活。
瑞士乳杆菌LBK-16H还表现出优异的胆汁耐受性,它耐受浓度高达0.5%的胆汁。该结果也显示出,该菌株可以在通过胃和小肠后在消化道中存活,并且在结肠中仍可存活。
而且,已经证明瑞士乳杆菌LBK-16H具有产生氧化氮的能力。对一氧化氮产生的研究发现,一氧化氮的形成是由于一氧化氮合成酶的激活导致的。在J774巨噬细胞系中,该菌株产生0.4μM的NO,而在T84人肠细胞系中该菌株产生约2-6μM的NO。该性质极有意义:激活产生适量的NO是有利的,例如在炎症反应和血压调节过程中。
还发现瑞士乳杆菌LBK-16H具有产生生物活性肽的能力。在芬兰专利申请992360的实施例中描述了如何通过两步骤方法制备易于保存的含有抗高血压肽的产品,在该方法的第一步中通过单独用瑞士乳杆菌LBK-16H或联合使用所述菌株和其它乳酸菌菌株发酵,例如各种乳制品,产生上述生物学活性肽。根据此公开文本的实施例,瑞士乳杆菌LBK-16H分别产生了已知的抗高血压三肽VPP(Val-Pro-Pro)和IPP(Ile-Pro-Pro)13-15mg/ml和6-8mg/ml。该公开文本指出,在发酵反应中形成了含有不同肽的混合物。当发酵的持续时间足够长时,获得的二和三肽相对较小。
基于上述特性,可以认为瑞士乳杆菌LBK-16H是一种益生生物。
瑞士乳杆菌LBK-16H是有用的,例如它可在乳品工业中,如在发酵乳制品和干酪的制备中,用作常规种子细菌。在干酪尤其是Emmental干酪的制备中的应用被认为是一个优选实施方案。
瑞士乳杆菌LBK-16H还可以极好地被用于制备含有生物活性肽的特殊产品,如发酵的乳制品,尤其是酸奶和酸乳酪。
瑞士乳杆菌LBK-16H还可以就这样用作功能性物质、或用于可食用物质的制备中,或作为可食用物质的成分或添加剂。
例如,可以在如MRS液体培养基中、在乳如生牛乳、再生的奶粉乳或经超声处理的乳中、或在通常用于乳酸菌属的培养基如Rogosa或MRS中,采用常规方法培养瑞士乳杆菌LBK-16H以制备该细菌。适合培养条件和其它参数,如温度、pH和通氧量,的选择是本领域技术人员所明了的。例如,温度可以是30-45℃。可能需要调节pH。
可以单独培养瑞士乳杆菌LBK-16H以形成纯培养物。也可以以混合培养物的形式培养该菌株,例如与本领域已知的其它发酵剂微生物一起培养。如果需要的话,还可以通过单独培养不同类型的微生物、然后按所需的比例将这些不同微生物组合在一起,以产生混合培养物。适当地选择该微生物组合,以便在终产品中提供最佳的可能性质并消除污染的危险。
培养后,回收细胞悬液,然后就这样使用,或以所需的方式例如通过浓缩、干燥或冻干进行处理。
自然,还可以以纯培养物或混合培养物的形式,单独地或例如与常规使用的和商业可获得的起子菌或益生菌一起使用瑞士乳杆菌LBK-16H。
根据本发明,在适当佐剂中,经冻干并含有菌株瑞士乳杆菌LBK-16H,DSM13137的细菌制品被认为是优选制品。
本发明的细菌制品可以就含有以上细菌或还含有其它组成成分如其它微生物。
根据本发明,该菌株还可以用于生产可食用产品,尤其是(功能性)食物、天然产品或药物。
本发明的可食用产品是通过使用瑞士乳杆菌LBK-16H,DSM13137、或含有该菌株的细菌产物和(终)产品的常规成分进行制备的。可以将该细菌在生产过程中加入食品或其它产品中,或将该菌株加入终产品中。还可以在产品的生产中使用该细菌,以便终产品中仅保留细菌生长过程中产生的产物,例如调味剂和芳香物质或具有生物活性的物质,而没有细菌细胞。
在本文中,术语食品具有广泛的含义,包括可以是固体、凝胶或液体形式的所有可食用产品,并包括速食产品和食用时加入了本发明产品作为添加剂或产品的组成成分的产品。例如,食品可以是乳品工业、肉类加工业、食品加工业、饮料工业、面包工业和糖果工业的产品。典型的产品包括乳制品如脱脂乳、具有不同脂肪含量的乳等(它们可以就这样存在或以相应的乳粉形式存在)、和发酵的乳制品如酸奶、酪乳、凝乳酪、酸乳酪、凝乳、未成熟干酪及成熟干酪、点心饼馅等。饮料如乳清饮料、水果饮料和啤酒构成了另一个重要的组。
对于本发明,制药业的产品除了包括各种药物制品外,还包括促进健康的天然产品等。制品的典型形式是例如胶囊、片剂和溶液等。
根据本发明,使用足够量的瑞士乳杆菌LBK-16H,DSM13137,以产生所需的效果。因此使用量可以根据用途和效果在一个宽范围内变化。
以下将通过实施例更为详细地描述本发明。这些实施例仅旨在例证本发明,而非以任何方式限制其范围。
实施例1
菌株瑞士乳杆菌LBK-16H的培养
通过在MRS或牛奶液体培养基中以1%的接种量接种保存菌株两次,37℃下培养菌株瑞士乳杆菌LBK-16H 20-24个小时。实际(生产性)也在基于MRS或基于乳的培养液中、于35℃-42℃培养该菌株20-48小时进行培养(调节或不调节pH)。
可以就这样从培养物中回收细胞,或可以通过利用已知技术将这些细胞浓缩并作为细胞浓缩物使用,或在浓缩后还可以将其冻干成粉末样产物。
实施例2
菌株瑞士乳杆菌LBK-16H的蛋白水解活性
通过OPA方法(Church,F.C.,Swaisgood,H.E.,Porter,D.H.,Catigna,G.L.,1983,使用邻苯二醛在牛奶和分离的乳蛋白中蛋白水解活性的分光光度测定法,J.Dairy Sci66:1219-1227),从牛奶中确定菌株瑞士乳杆菌LBK-16H的蛋白水解活性。
为了实施该方法,将细菌细胞培养在含有MRS液体培养基的10ml试管中,之后用0.9%NaCl洗涤悬浮物,并重悬至10ml。按1%的接种量将细菌细胞接种在10ml 22度的奶中。使用不含细菌接种物的相应乳样品作为对照。
在涡旋装置中充分混合2.5ml样品、0.5ml水和0.5ml 0.75N三氯乙酸(TCA),使其静置10分钟,然后通过Whatman 2号滤纸过滤。从TCA过滤物中取150μl样品,并与3ml OPA试剂在一次性杯中混合。室温孵育混合物2分钟,之后用分光光度计在340nm波长下进行测量。从读数中减去对照值,由此瑞士乳杆菌LBK-16H的蛋白水解活性为0.3-0.6。
OPA试剂:
25ml 100mmol偏硼酸钠
2.5ml 20%(w/w)SDS
40mg溶解在1ml甲醇中的OPA(邻苯二醛)
100μl β-巯基乙醇
将这些物质混合并溶解在水中,终体积为50ml。该试剂储存在暗处可保存1天。
实施例3
瑞士乳杆菌LBK-16H的pH耐受性
测试前两次在MRS液体培养基中培养LBK-16H 17-18小时。之后,在MRS液体培养基或牛奶液体培养基中培养该菌株以便测试。在pH测试中,将测试液体培养基(MRS)的pH调节至pH6.5、pH4和pH2(用乳酸)。用MRS或牛奶液体培养基的培养物接种这些液体培养基,以便初始浓度为107cfu/ml,并在37℃培养3小时和4小时,之后测定这些培养物中菌株LBK-16H的浓度(在MRS琼脂上通过乳酸菌属的测定方法测定)。结果见表1。表1.菌株LBK-16H的pH耐受性
因此该菌株极好地耐受4甚至3的pH水平4小时。与加入水性产品中相比,当加入至乳制品中时该菌株在低至2的pH水平上4小时的保存性更好。
3小时 4小时 |
初始浓 pH pH4 pH3 pH pH4 pH3 pH2度 6.5 6.5 |
MRS 4×107 6×107 3×107 5×107 6×107 3×107 3×107 <100 |
乳 5×107 5×107 3×107 6×107 5×107 4×107 7×107 1×104 |
实施例4
菌株瑞士乳杆菌LBK-16H的胆汁耐受性
按实施例3培养LBK-16H。在加有0.3%和0.5%胆汁酸(牛胆汁,SigmaB-3883)的MRS液体培养基中进行测试。在该含有胆汁的液体培养基中加入1%新鲜培养物,初始浓度为2-5×106cfu/ml,其中该新鲜培养物来自基于MRS或牛奶的培养物。在该液体培养基中培养该菌株3小时,之后在MRS琼脂上测定细胞的浓度。结果见表2。表2.LBK-16H的胆汁耐受性
生长培养基 | 胆汁浓度0 0.3% 0.5% |
MRS奶 | 2×106 2×106 4×1055×106 6×106 5×106 |
正如该表显示的,LBK-16H耐受浓度高达0.5%的胆汁。
实施例5
由瑞士乳杆菌LBK-16H产生的一氧化氮
采用Korhonen等描述的方法(在J774巨噬细胞和T84结肠上皮细胞中通过益生乳酸菌GG诱导一氧化氮合成,由Korhonen,R.,Korpela,R.,Saxelin,M,Maki,M.,Kankaanranta,H.和Moilanen,E.已提交),研究由瑞士乳杆菌LBK-16H引起的硝酸产生。该方法是基于诱导型一氧化氮合成酶(iNOS)的诱导和所导致的一氧化氮(NO)的产生。在本试验中采用两种不同的细胞系:J774小鼠巨噬细胞系和T84人肠细胞系。由于单独的细菌细胞并不产生NO,所以在存在γ-干扰素时用瑞士乳杆菌LBK-16H细胞在J774细胞系中进行诱导。该细胞系细胞和细菌株细胞的细胞比例是1∶10。
采用脂多糖(LPS)和脂磷壁酸质(LA)作为参考,通过Western印迹技术确定酶iNOS的表达。
以24小时孵育后生长培养基中一氧化氮代谢物亚硝酸盐的量,确定一氧化氮(NO)的产生。亚硝酸盐的量通过Griess反应(Green等,1988,分析生物化学,第126卷,第131-138页)来测量。
瑞士乳杆菌LBK-16H能够在小鼠巨噬细胞系中产生0.4μM的一氧化氮,在人上皮细胞系中产生5-6μM一氧化氮。与此比较,Lactobacillusrhamonosus LC705,DSM7061,在T84细胞中仅产生0.7μM一氧化氮。实施例6
由瑞士乳杆菌LBK-16H产生的生物活性肽
在MRS液体培养基中37℃ 24小时培养菌株瑞士乳杆菌LBK-16H,并用该菌株接种再生乳(10%)以便形成接种物。两轮培养后,将接种物(15%)接种在发酵培养基中,该培养基由9-10%脱脂奶粉奶组成,并在110℃灭菌10min。发酵在37℃进行22-24小时,始终都进行有力的混合。
为了进行比较试验,采用(a)几种菌株,即瑞士乳杆菌LB161、瑞士乳杆菌LBK-16H和瑞士乳杆菌LB230的混合物,(b)菌株瑞士乳杆菌LBK-16H和鼠李糖乳杆菌LC705,DSM7061的混合物,和(c)菌株瑞士乳杆菌LBK-16H和嗜热链球菌(Streptococcus thermophilus)T101,DSM4022的混合物,重复该发酵实验。
所用生长培养基是在100℃灭菌15分钟的9%乳。为了形成接种物,将菌株瑞士乳杆菌和鼠李糖乳杆菌LC705在MRS液体培养基中37℃培养24小时,然后从其中转移1%的接种物至乳中。嗜热链球菌T101在LM17液体培养基中37℃培养18小时,然后从其中将接种物转移至乳中。
第一次培养通过在乳中37℃孵育24小时,单独培养所有菌株。对于第二次培养,将1%的每种混合物的菌株转移至乳中,之后37℃继续共培养24小时。对于第三次培养,将5-10%的上述所获得的共培养物转移至乳中,并37℃孵育24小时。
表3显示了菌株瑞士乳杆菌LBK-16H和这些不同微生物混合物产生的VPP和IPP的量。本发明的瑞士乳杆菌LBK-16H能够产生大量的生物活性肽。这些不同混合物中的其它微生物均不产生生物活性肽,但它们也不干扰LBK-16H的活性。在混合物之间或单独考虑菌株LBK-16H,没有观察到差异。
表3
菌株LBK-16H和不同的微生物混合物产生的IPP和VPP的量
微生物(混合物) | VPP,mg/l | IPP,mg/l |
L161+LBK-16H+LB230,5%L161+LBK-16H+LB230,10%LBK16H+LC705,5%LBK16H+LC705,10%LBK16H+Str.T101 10%LBK16H(单独地) | 13-1413-1413-1413-1413-1513-15 | 6-76-76-76-76-86-8 |
Claims (14)
1.瑞士乳杆菌LBK-16H,DSM13137。
2.含有菌株瑞士乳杆菌LBK-16H,DSM13137的细菌制品。
3.权利要求2的细菌制品,其特征在于还含有其它微生物。
4.权利要求2或3的细菌制品,其特征在于是冻干的粉末或胶囊形式。
5.菌株瑞士乳杆菌LBK-16H,DSM13137在食品工业中的用途。
6.权利要求5的用途,其特征在于制备的是乳品工业或饮料工业的产品。
7.可食用产品,其特征在于含有权利要求1的细菌或权利要求2-4之任一项的细菌制品,或是采用它们制备的。
8.权利要求7的可食用产品,其特征在于是乳制品。
9.权利要求7的可食用产品,其特征在于是饮料,优选乳清饮料、水果饮料或啤酒。
10.菌株瑞士乳杆菌LBK-16H,DSM13137,作为治疗性物质的用途。
11.瑞士乳杆菌LBK-16H,DSM13137,在高血压治疗中的用途。
12.菌株瑞士乳杆菌LBK-16H,DSM13137,在制备抗高血压产品中的用途。
13.权利要求12的用途,其特征在于该抗高血压产品含有抗高血压二和三肽。
14.权利要求12的产品,其特征在于具有高浓度的三肽,尤其是Ile-Pro-Pro和/或Val-Pro-Pro。
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Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
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CN100402089C (zh) * | 2003-03-18 | 2008-07-16 | 三得利株式会社 | 血管紧张素转化酶抑制肽 |
CN100370019C (zh) * | 2006-04-21 | 2008-02-20 | 江南大学 | 一种具有抗高血压作用的植物乳杆菌cw006 |
CN101678051B (zh) * | 2006-09-04 | 2012-12-12 | 雪印惠乳业株式会社 | 促进血中脂连蛋白浓度增加及/或抑制其减少的制剂及内脏脂肪蓄积抑制剂 |
CN102786580A (zh) * | 2012-07-17 | 2012-11-21 | 上海交通大学 | 抗高血压活性肽ivp |
CN102786579A (zh) * | 2012-07-17 | 2012-11-21 | 上海交通大学 | 抗高血压活性肽vip |
CN102786579B (zh) * | 2012-07-17 | 2014-07-09 | 上海交通大学 | 抗高血压活性肽vip |
CN109517763A (zh) * | 2018-12-27 | 2019-03-26 | 内蒙古农业大学 | 瑞士乳杆菌h11及其应用 |
CN109517763B (zh) * | 2018-12-27 | 2022-03-22 | 内蒙古农业大学 | 瑞士乳杆菌h11及其应用 |
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