CN101616680B - 内脏脂肪减少剂 - Google Patents
内脏脂肪减少剂 Download PDFInfo
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- CN101616680B CN101616680B CN200880005463XA CN200880005463A CN101616680B CN 101616680 B CN101616680 B CN 101616680B CN 200880005463X A CN200880005463X A CN 200880005463XA CN 200880005463 A CN200880005463 A CN 200880005463A CN 101616680 B CN101616680 B CN 101616680B
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Abstract
本发明提供以乳酸菌、特别是乳杆菌属及乳球菌属的乳酸菌的菌体及/或其培养物为有效成分,用于减轻作为高血压、高血脂症、糖耐量异常等危险因素聚集后的状态的代谢综合征的病状、减少内脏脂肪积累的内脏脂肪减少剂、含有该内脏脂肪减少剂的饮食品或饲料。
Description
技术领域
本发明涉及以乳酸菌、特别是乳杆菌属及乳球菌属乳酸菌的菌体及/或其培养物为有效成分的内脏脂肪减少剂。
此外,本发明还涉及通过配合上述内脏脂肪减少剂,从而赋予了减少内脏脂肪作用的新型饮食品或饲料。
内脏脂肪的积累是与血栓症、胰岛素抵抗、糖代谢异常、高血压等未来循环系统疾病的发病有关的所谓代谢综合征的原因,通过摄取本发明的内脏脂肪减少剂,可减少积累的内脏脂肪,有效地预防·治疗代谢综合征。
背景技术
近年来,随着生活习惯的欧美化,呈现作为生活习惯病的糖尿病、高血压、高血脂症、动脉硬化症病状的人口正在增加。特别是因心血管疾病和脑血管疾病引起的死亡约占死因的3分之1,且其数量在逐年增加,所以,针对这些的对策已成为国民性的课题。这些动脉硬化性疾病随着高血压、高血脂症、糖耐量异常等危险因素的聚集而使发病的危险度显著上升。此危险因素聚集的状态被称为代谢综合征,并已被广泛认识。
由对日本的12万企业劳动者的调查可知,即使是具有症状较轻的“肥胖”、“高血压”、“高血糖”、“高甘油三酯(中性脂肪)血症”、或“高胆固醇血症”中的危险因素中的1项的人的心脏病发病危险也为5倍,具有2项的人为10倍,同时具有3~4项的人为31倍。而且,在厚生劳动省的调查中,报道了高血压患者的数量为3,900万人、高血脂症为2,200万人、糖尿病(包含预备军)为1,620万人、肥胖症为468万人,这些患者还在逐年增加。
代谢综合征是指“内脏脂肪的积累、和在此基础上并发胰岛素抵抗性及糖代谢异常、脂质代谢异常、高血压多项的多重危险因素综合征,是易患动脉硬化的病状”,因而内脏脂肪的积累无疑是其根本性原因。作为生物体最大分泌组织的脂肪组织产生各种内分泌因子,与维持生物体的恒常性有关。
但是,现在已经了解了内脏脂肪的过度积累会破坏内分泌因子的分泌平衡,而引发各种病状。特别是纤溶酶原激活物抑制物(PAI-1)、肿瘤坏死因子(TNF-α)、瘦蛋白等的内分泌因子随内脏脂肪的积累而分泌量增加,从而引起血栓症、胰岛素抵抗性、糖代谢异常、高血压等。
另一方面,已知脂肪组织特异性分泌的脂联素通常在血液中高浓度存在,但随内脏脂肪的积累其浓度减少。已知脂联素具有抗糖尿病、抗动脉硬化、抗炎症作用、抗高血压等各种生理功能,在血液中促进脂联素浓度的增加、或抑制脂联素浓度的减少对预防·治疗代谢综合征非常重要。
以往,作为针对代谢综合征的各种病状的对策进行过药物疗法,但具有需要处方和伴随副作用等的问题。而且还可知,针对一种病状进行治疗时,其他病状却有机会而向严重病状发展,所以需要调整在这种状态的上游存在的来自脂肪细胞的内分泌因子的分泌平衡。
因此,在由内脏脂肪的积累引起的代谢综合征的预防·治疗中,与药物疗法相比,采取运动疗法、饮食疗法等改善每天的生活方式更为重要。因此,希望有可日常摄取、即使长期摄取安全性也高、对内脏脂肪积累所引起的代谢综合征的预防·治疗有效的药剂、饮食品或饲料。
专利文献1中公开了作为对人体有效的成分的乳酸菌的适用:属于被分类为嗜酸乳杆菌(Lactobacillus acidophilus)、卷曲乳杆菌(Lactobacilluscrispatus)、食淀奶粉杆菌(Lactobacillus amylovorus)、格氏乳杆菌(Lactobacillus gasseri)、鸡乳杆菌(Lactobacillus gallinarum)及约氏乳杆菌(Lactobacillus johnsonii)的嗜酸乳杆菌复合物(Lactobacillus acidophiluscomplex),与唾液酸基缺乏(asialo)GM1有亲和性的乳酸菌及其提取物、对与唾液酸基缺乏(asialo)GM1有亲和性的病原体的感染预防有效。
此外,在专利文献2中记载了以格氏乳杆菌(Lactobacillus gasseri)的菌体或其发酵产物为有效成分的炎性肠疾病及过敏性肠综合征的预防·治疗剂有效。而且在专利文献3中还公开了以属于格氏乳杆菌(Lactobacillus gasseri)的乳酸菌及其培养物、和属于长双歧杆菌(Bifidobacterium longum)的乳酸菌及其培养物为有效成分的糖尿病并发症的预防·改善·治疗剂。
作为以培养属于格氏乳杆菌(Lactobacillus gasseri)的乳酸菌而得到的菌体及培养物为有效成分的制剂,除上述以外,专利文献4中还公开了血清胆固醇上升抑制剂。此外,专利文献5中还记载了以培养同样属于格氏乳杆菌(Lactobacillus gasseri)的乳酸菌而得到的菌体及其培养物为有效成分的骨吸收抑制剂,公开了具有骨吸收抑制作用的饮食品或饲料。专利文献6中公开了不是单独使用属于格氏乳杆菌(Lactobacillus gasseri)的乳酸菌,而是将燕麦等的禾本科谷粒及其加工物与属于格氏乳杆菌(Lactobacillus gasseri)的乳酸菌组合用于增强免疫的免疫增强剂及免疫增强用饮食品,其显示出与各个单独使用相比,巨噬细胞被激活,细胞因子产生量协同增高,增大了免疫增强效果。但是,格氏乳杆菌(Lactobacillusgasseri)的培养物、或其菌体自身具有减少内脏脂肪作用还不为人所知。
而且,从古至今就作为代表性的乳业乳制品用乳酸菌发酵剂使用、且作为乳杆菌属(Lactobacillus sp.)的乳酸菌的瑞士乳杆菌(Lactobacillushelveticus)具有强的蛋白水解活性,特别是具有高活性的菌体外蛋白酶对乳品发酵性起着重要的作用。即菌体外蛋白酶水解乳蛋白质,生成各种肽片段。生成的肽受肽酶组的作用,形成更低分子的肽。已知通过蛋白质水解酶组的作用而在培养基中生成的肽的一部分被摄入乳酸菌的菌体内,作为氮源利用。
另一方面,非专利文献1中报到了在培养基中生成的肽中,存在对于作为升压作用的原因物质的血管紧张肽转换酶(Angiotensin ConvertingEnzyme,以下称ACE)具有抑制活性的肽。对于以抑制ACE的酶活性、抑制血压上升为目的的肽,非专利文献2中已报道了乳蛋白质、大豆蛋白质或鱼肉蛋白水解物等中的多种有效肽,公开了使用了瑞士乳杆菌(Lactobacillus helveticus)的发酵乳中具有ACE抑制活性的肽是Val-Pro-Pro和Ile-Pro-Pro,且这些lactotripeptide具有强降压作用,该结论已通过使用自然发病的高血压大鼠(SHR)的实验得到了证实。这样,含有由瑞士乳杆菌(Lactobacillus helveticus)发酵的酸奶的功能性食品不仅具有降压活性,还具有缓解精神压力的作用,这已由专利文献7及8进行了阐明。此外,专利文献9中还揭示了瑞士乳杆菌(Lactobacillushelveticus)、干酪乳杆菌(Lactobacillus casei)、德氏乳杆菌保加利亚亚种(L.delbrueckii subsp,bulgaricus)、德氏乳杆菌乳酸亚种(L.delbrueckiisubsp.lactis)等的乳酸菌具有免疫赋活作用。
但是,这些乳酸菌的菌体自身或其培养物具有减少内脏脂肪作用,还不为人所知。
乳酸乳球菌(Lactococcus lactis)及乳脂乳球菌(Lactococcus cremoris)是乳制品制造中广泛使用的乳酸菌,但有关其生理功能还知之甚少,虽已知用乳球菌属乳酸菌使牛乳、豆乳、咸菜等发酵时能产生γ氨基丁酸(GABA)等,具有调节血压的作用等,但其有助于内脏脂肪的减少还不为人所知。
专利文献1日本特开平8-268899号公报
专利文献2日本特开2003-95963号公报
专利文献3日本特开2003-252770号公报
专利文献4日本特开2003-306436号公报
专利文献5日本特开2004-315477号公报
专利文献6日本特开2006-69993号公报
专利文献7日本特开平10-45610号公报
专利文献8日本特开平11-98978号公报
专利文献9日本特开2006-76961号公报
非专利文献1J.Dairy Sci.,78:777-783(1995)
非专利文献2J.Dairy Sci.,78:1253-1257(1995)
发明内容
本发明的课题是提供可日常摄取、且通过摄取使内脏脂肪减少从而对代谢综合征的预防·治疗有效的内脏脂肪减少剂、及赋与了该功能的饮食品或饲料。
本发明者对乳成分中使引起代谢综合征的内脏脂肪降低的成分进行锐意研究,其结果,在乳酸菌、特别是乳杆菌属(Lactobacillus sp.)及乳球菌属(Lactococcus sp.)乳酸菌中,发现了极强的减少内脏脂肪作用,从而完成了本发明。
已知上述乳杆菌属及乳球菌属的乳酸菌可用于发酵乳、奶酪、咸菜物等各种发酵食品的制造,是从乳业乳酸菌、人体消化道的肠道菌群中分离的乳酸菌,不仅具有调节肠胃功能的作用,还具有免疫调节作用、降低血清胆固醇作用、预防感染作用等多种生理功能,而本发明中指以乳杆菌属及乳球菌属乳酸菌的菌体及/或其培养物为有效成分的内脏脂肪减少剂及赋与了该功能的饮食品或饲料。
上述内脏脂肪减少剂的使用方式有片剂、散剂、胶囊剂等。
且其使用用途的指定考虑为发酵乳、健康食品或奶酪类等的饮食品或饲料等。
本发明原则上是由以下构成形成的发明。
(1)内脏脂肪减少剂,其以乳酸菌的菌体及/或其培养物为有效成分。
(2)根据上述(1)中的内脏脂肪减少剂,其特征在于,以乳杆菌属及/或乳球菌属乳酸菌的菌体及/或其培养物为有效成分。
(3)根据上述(2)中的内脏脂肪减少剂,其特征在于,以格氏乳杆菌(Lactobacillus gasseri)、瑞士乳杆菌(Lactobacillus helveticus)、乳酸乳球菌(Lactococcus lactis)、乳脂乳球菌(Lactococcus cremoris)中的任一种以上的菌体及/或其培养物为有效成分。
(4)减少内脏脂肪用片剂、散剂、胶囊剂、饮食品或饲料,其特征在于,配合了上述(1)~(3)中任一项所述的乳酸菌的菌体及/或其培养物。
(5)根据上述(4)中的饮食品,其特征在于,上述饮食品为发酵乳、健康食品或奶酪类等。
本发明的以乳酸菌的菌体及/或其培养物为有效成分的内脏脂肪减少剂、及含有该内脏脂肪减少剂且赋与了减少内脏脂肪作用的饮食品或饲料对因内脏脂肪积累而发病的所谓代谢综合征的预防·治疗有效。
且本发明的内脏脂肪减少剂、及赋与了减少内脏脂肪作用的饮食品或饲料,因使用乳杆菌属(Lactobacillus sp.)乳酸菌及乳球菌属(Lactococcussp.)乳酸菌、特别是其中的格氏乳杆菌(Lactobacillus gasseri)、瑞士乳杆菌(Lactobacillus helveticus)、乳酸乳球菌(Lactococcus lactis)或乳脂乳球菌(Lactococcus cremoris)的培养物或其菌体自身,所以可比较廉价、大量地供给,且具有安全性极高的效果。
具体实施方式
作为本发明中的乳酸菌,可例举属于乳杆菌属(Lactobacillus sp.)的乳酸菌及乳球菌属(Lactococcus sp.)的乳酸菌、特别是其中的格氏乳杆菌(Lactobacillus gasseri)、瑞士乳杆菌(Lactobacillus helveticus)、乳酸乳球菌(Lactococcus lactis)或乳脂乳球菌(Lactococcus cremoris)的乳酸菌。
可选择属于格氏乳杆菌(Lactobacillus gasseri)的乳酸菌中的格氏乳杆菌(Lactobacillus gasseri)SBT2055株、格氏乳杆菌(Lactobacillus gasseri)JCM1131株及格氏乳杆菌(Lactobacillus gasseri)ATCC19992株。
且SBT2055株的保藏接收编号为FERM ABP-10953,保藏于日本的独立行政法人产业技术综合研究所专利生物保藏中心(茨城县筑波市东1丁目1番地1中央第6、接收日;2008年2月26日)。
另外,JCM1131株是独立行政法人理化学研究所的公开菌株,ATCC19992株是美国菌种保藏中心(American Type Culture Collection)的公开菌株。
而且,可选择属于瑞士乳杆菌(Lactobacillus helveticus)的乳酸菌中的瑞士乳杆菌(Lactobacillus helveticus)SBT2171株、瑞士乳杆菌(Lactobacillus helveticus)ATCC10386株及瑞士乳杆菌(Lactobacillushelveticus)ATCC 10797株。
此外,SBT2171株的保藏编号为FERM BP-5445,保藏于独立行政法人产业技术综合研究所专利生物保藏中心(茨城县筑波市东1丁目1番地1中央第6、接收日;1996年3月6日)。且ATCC10386株及ATCC10797株为美国菌种保藏中心(American Type Culture Collection)的公开菌株。
而且,还可选择属于乳酸乳球菌(Lactococcus lactis)的乳酸菌中的乳酸乳球菌(Lactococcus lactis)JCM1107株。乳酸乳球菌(Lactococcuslactis)JCM1107株为独立行政法人理化学研究所的公开菌株。
进而还可选择属于乳脂乳球菌(Lactococcus cremoris)的乳酸菌中的乳脂乳球菌(Lactococcus cremoris)ATCC11602株、乳脂乳球菌(Lactococcus cremoris)ATCC14365株、乳脂乳球菌(Lactococcus cremoris)ATCC19257株。此外,ATCC11602株、ATCC14365株及ATCC19257株为美国菌种保藏中心(American Type Culture Collection)的公开菌株。
且对于乳酸菌的筛选,随着逐步筛选耐胃酸性高、低pH条件下生育良好、对人体肠道显示高定居性、与人体肠道细胞显示高度亲和性、有耐胆汁酸性、在肠道内定居、适用于食品时存活性高、香味、物性也优异的菌株,可筛选出更适合本发明的乳酸菌。
特别是格氏乳杆菌(Lactobacillus gasseri)SBT2055株(FERMABP-10953)与人体肠道细胞有高度亲和性,经口给药时,能以存活状态到达肠道内,且可长时间常驻于肠道内,通过在肠道内生长发育而对宿主起作用,具有减少内脏脂肪效果的活性高。对从体外投给的属于格氏乳杆菌(Lactobacillus gasseri)的乳酸菌在肠内定居而显示上述生理效果还全然不知,由本发明者首次查明了这一点。
而且在本发明中,不限于上述菌株,即使是从人体、发酵乳中分离的格氏乳杆菌(Lactobacillus gasseri),不仅是活菌体,即使是死菌体,只要显示上述作用,均可使用。此外,也可使用作为格氏乳杆菌(Lactobacillusgasseri)的突变株、显示上述作用的菌株。
即使是上述乳酸菌中的格氏乳杆菌(Lactobacillus gasseri)以外的乳酸菌,与格氏乳杆菌(Lactobacillus gasseri)同样被称作嗜酸乳杆菌(Lactobacillus acidophilus)群的卷曲乳杆菌(Lactobacillus crispatus)、作为高温性同型的保加利亚乳杆菌(Lactobacillus delbrueckii subsp.Bulgaricus)等的乳品加工用乳酸菌、作为中温性同型的乳酸菌的植物乳杆菌(Lactobacillus plantarum)、干酪乳杆菌(Lactobacillus casei)等也具有同样的减少内脏脂肪的效果。
培养作为本发明的乳酸菌的格氏乳杆菌(Lactobacillus gasseri)的培养基,可使用乳培养基或含有乳成分的培养基、不含有该成分的半合成培养基等的各种培养基。作为此种培养基,可例示还原脱脂乳后加热杀菌的还原脱脂乳培养基。
这些乳杆菌属及乳球菌属乳酸菌的培养法,为进行静置培养或进行将pH控制为恒定的中和培养,但只要是菌体良好生长发育的条件,对培养法无特别限制。菌体根据乳酸菌培养的常法进行培养,可将从所得到的培养物中通过离心分离等的集菌方法分离后的产物直接作为本发明的有效成分使用。此外,作为菌体,不仅可使用单纯分离的菌体,还可使用培养物、悬浮物、其他的菌体含有物、使用酶、物理手段处理菌体后的细胞质、细胞壁组分。而且,不仅可使用活菌体,也可使用死菌体。
本发明的内脏脂肪减少剂以乳杆菌属(Lactobacillus sp.)及乳球菌属(Lactococcus sp.)的乳酸菌的培养物及/或其菌体自身为有效成分。而且,在制备制剂时,除可适当配合制剂上允许的赋形剂、稳定剂、矫味剂等后进行浓缩、冷冻干燥以外,还可以加热干燥成死菌体。也包含这些的干燥物、浓缩物、糊状物。此外,在不妨碍菌体、培养物的作用的范围内,还可混合赋型剂、粘合剂、崩解剂、润滑剂、矫味矫臭剂、悬浮剂、包衣剂、其他任意的药剂来制备制剂。作为剂形,可为片剂、丸剂、胶囊剂、颗粒剂、散剂、粉剂、糖浆剂等,并希望将这些经口给药。
此外,本发明还指以乳杆菌属(Lactobacillus sp.)乳酸菌及乳球菌属(Lactococcus sp.)的乳酸菌、特别是其中的格氏乳杆菌(Lactobacillusgasseri)、瑞士乳杆菌(Lactobacillus helveticus)、乳酸乳球菌(Lactococcuslactis)或乳脂乳球菌(Lactococcus cremoris)的培养物、或其菌体自身为有效成分并赋与了减少内脏脂肪功能的饮食品或饲料。作为本发明的赋与了减少内脏脂肪作用的饮食品或饲料,可以是乳杆菌属(Lactobacillus sp.)乳酸菌及乳球菌属(Lactococcus sp.)乳酸菌、特别是其中的格氏乳杆菌(Lactobacillus gasseri)、瑞士乳杆菌(Lactobacillus helveticus)、乳酸乳球菌(Lactococcus lactis)或乳脂乳球菌(Lactococcus cremoris)的培养物或其菌体自身,也可以是配合了菌体、培养物自身的饮食品。作为培养物的形态,酸奶、奶酪等的发酵食品均为适合形态。
菌体、培养物可配合在任意饮食品中,也可在饮食品的制造过程中添加到原料中。做为饮食品的例子,可例举乳饮料、发酵乳、果汁饮料、果冻、糖果、乳制品、沙拉酱等的蛋类加工品、黄油蛋糕等的点心·面包类等的食品。此外,除各种奶粉以外,还可例示配合在以婴儿、幼儿及低出生体重儿等为对象的营养组合物中的饮食品。作为活菌体利用时,作为材料也可使用乳杆菌属(Lactobacillus sp.)乳酸菌及乳球菌属(Lactococcussp.)乳酸菌、特别是其中的格氏乳杆菌(Lactobacillus gasseri)、瑞士乳杆菌(Lactobacillus helveticus)、乳酸乳球菌(Lactococcus lactis)或乳脂乳球菌(Lactococcus cremoris)的菌株自身、及使上述乳酸菌的菌体发酵后得到的发酵乳、奶酪,制成用于减少内脏脂肪的面包、休闲食品、蛋糕、布丁等。因这些饮食品可日常摄取且具有减少内脏脂肪的作用,所以对代谢综合征的预防·治疗有效。
本发明还进一步指以乳杆菌属(Lactobacillus sp.)乳酸菌及乳球菌属(Lactococcus sp.)乳酸菌、特别是其中的格氏乳杆菌(Lactobacillusgasseri)、瑞士乳杆菌(Lactobacillus helveticus)、乳酸乳球菌(Lactococcuslactis)或乳脂乳球菌(Lactococcus cremoris)的培养物、或其菌体自身为有效成分并赋与了减少内脏脂肪功能的饲料。作为家畜用饲料,与所述饮食品同样,菌体、培养物可与任何饲料配合,也可在饮食品的制造过程中添加到原料中。
在本发明中,为发挥减少内脏脂肪作用,成人的情况下,可将乳杆菌属(Lactobacillus sp.)乳酸菌及乳球菌属(Lactococcus sp.)乳酸菌、特别是其中的格氏乳杆菌(Lactobacillus gasseri)、瑞士乳杆菌(Lactobacillushelveticus)、乳酸乳球菌(Lactococcus lactis)或乳脂乳球菌(Lactococcuscremoris)的培养物调节到每天可摄取10~200g、或调节到每天可摄取其菌体自身0.1~5,000mg的配合量等。乳酸菌的含有比例无特别限定,可结合制造的容易性、优选的每天给药量等适当调节。例如剂型为液体时,优选乳酸菌体浓度为1×105cells/ml~1×1010cells/ml,为固体时,优选为1×105cells/g~1×1010cells/g。作为活菌给药时,成人每人以108~1012cfu/日给药,即可发挥本发明的目的效果。通过如此摄取,可使乳酸菌在肠道内定居并发挥所需的效果。
以乳杆菌属的乳酸菌的菌体及其培养物为有效成分的内脏脂肪积累抑制剂及赋与了抑制内脏脂肪积累作用的饮食品(日本特愿2006-239290号)已由本申请人进行了申请,但其是以抑制内脏脂肪本身的积累为目的的发明,而其中未发现如本发明中的内脏脂肪积累后的减少作用。
此外,本发明不仅具有更迅速且有效地改善代谢综合征的各种病状的特征,而且在属于乳球菌属的乳酸菌的菌体及其培养物中也发现了有效成分。
因本发明中的内脏脂肪减少剂、或减少内脏脂肪用饮食品、饲料具有减少内脏脂肪的效果,所以,可非常有益于预防、治疗、改善由前述的内脏脂肪积累所引起的种种病状。
本发明中使用的乳酸菌体自古以来被用于发酵乳、奶酪的制造,所以,本发明的内脏脂肪减少剂、或减少内脏脂肪用饮食品、饲料具有不存在安全性问题的特征。
实施例
以下表示实施例及试验例,对本发明进行更加详细的说明,但这些只不过是单纯的举例,本发明不受这些的任何限定。
实施例1
(格氏乳杆菌培养物粉末的制备1)
将还原脱脂乳培养基(含有13重量%脱脂奶粉、0.5重量%酵母浸膏)在95℃下杀菌30分钟后,接种格氏乳杆菌(Lactobacillus gasseri)SBT2055(FERM ABP-10953),37℃下培养16小时,冷冻干燥所得到的培养物后,得到格氏乳杆菌(Lactobacillus gasseri)SBT2055(FERM ABP-10953)的培养物粉末。该培养物粉末可直接作为本发明的内脏脂肪减少剂使用。
实施例2
(格氏乳杆菌培养物粉末的制备2)
将还原脱脂乳培养基(含有13重量%脱脂奶粉、0.5重量%酵母浸膏)在95℃下杀菌30分钟后,接种格氏乳杆菌(Lactobacillus gasseri)JCM1131,37℃下培养16小时,冷冻干燥所得到的培养物后,得到格氏乳杆菌(Lactobacillus gasseri)JCM1131的培养物粉末。该培养物粉末可直接作为本发明的内脏脂肪减少剂使用。
实施例3
(片剂的制造)
将格氏乳杆菌(Lactobacillus gasseri)SBT2055(FERM ABP-10953)的液体培养物在4℃、7,000rpm的条件下离心分离15分钟后,重复进行3次用灭菌水洗涤和离心分离,得到洗涤菌体。将该洗涤菌体进行冷冻干燥处理后,得到菌体粉末。在该菌体粉末1份中混合脱脂奶粉4份,用压片机将该混合粉末通过常法按每片1g进行压片,从而制备本发明的含200mg格氏乳杆菌(Lactobacillus gasseri)SBT2055(FERM ABP-10953)的菌体的片剂。
实施例4
(发酵乳的制造)
将格氏乳杆菌(Lactobacillus gasseri)SBT2055(FERM ABP-10953)在MRS液体培养基(商品名Lactobacilli MRS Broth,Difco公司制品)中培养。将处于对数增殖期的各培养液在添加有0.3重量%的酵母浸膏的10重量%还原脱脂乳(115℃、20分钟灭菌后)中接种1重量%,制成母种培养液。在其中添加10重量%的还原脱脂乳,在100℃下加热10分钟的酸奶混合物中添加2.5重量%来进行制备。在37℃下进行发酵,在乳酸酸度达到0.85时进行冷却,使发酵结束,得到本发明的减少内脏脂肪用发酵乳。
实施例5
(散剂的制造)
将格氏乳杆菌(Lactobacillus gasseri)SBT2055(FERM ABP-10953)接种于5L MRS液体培养基(商品名Lactobacilli MRS Broth,Difco公司)中后,进行37℃下18小时的静置培养。培养结束后,进行7,000rpm、15分钟的离心分离,得到培养液的1/50量的浓缩菌体。而后,将该浓缩菌体与含有脱脂奶粉10重量%、谷氨酸钠1重量%的分散介质等量混合,pH调整到7后进行冷冻干燥。将所得到的冷冻干燥物用60目的筛子整粒,制造冷冻干燥菌粉。以第13改正日本药局方解说书制剂总则“散剂”的规定为依据,在该冷冻干燥菌粉1g中加入乳糖(日局)400g、马铃薯淀粉(日局)600g均匀混合,制得本发明的减少内脏脂肪用散剂。
实施例6
(棒状健康食品的制造)
在实施例1中得到的格氏乳杆菌(Lactobacillus gasseri)SBT2055(FERM ABP-10953)的培养物粉末30g中,加入维生素C和柠檬酸的等量混合物40g、精制白砂糖100g、玉米淀粉和乳糖的等量混合物60g后进行混合。将混合物装入棒状袋中,制成本发明的减少内脏脂肪用棒状健康食品。
实施例7
(天然奶酪的制造)
将调整了脂肪率的原料乳在75℃下进行15秒的片式加热杀菌后,冷却到30℃,添加0.01重量%氯化钙。而后,在这些原料乳中添加市售的乳酸菌发酵剂(科汉森(Chr.Hansen)公司制)0.7重量%及格氏乳杆菌(Lactobacillus gasseri)SBT2055(FERM ABP-10953)1重量%,添加凝乳酶0.003重量%使乳凝固后,切割并搅拌至pH为6.2~6.1,排出乳清,得到凝乳粒。再将此凝乳粒成型压榨,进一步盐渍,制成本发明的减少内脏脂肪用天然奶酪。
实施例8
(胶囊剂的制造)
按表1中所示的组成将原料混合、整粒后,填充到胶囊中,制成本发明的减少内脏脂肪用胶囊剂。
表1
实施例9
(饮料的制造)
按表2中所示的组成将原料混合、填充到容器中后,进行加热杀菌,制成本发明的减少内脏脂肪用饮料。
表2
实施例10
(低脂肪硬质天然奶酪的制造)
使用使奶酪中的脂肪率为12~30重量%而制备的原料乳,制成多种低脂肪硬质天然奶酪。即将调整了脂肪率的原料乳在75℃下进行15秒的片式加热杀菌后,冷却到30℃,添加0.01重量%氯化钙。而后,在这些原料乳中添加乳酸菌发酵剂(科汉森(Chr.Hansen)公司制)0.7重量%及瑞士乳杆菌(Lactobacillus helveticus)SBT2171(FERM BP-5445)1重量%,添加凝乳酶0.003重量%使乳凝固后,切割并搅拌至pH为6.2~6.1,排出乳清,得到凝乳粒。再将此凝乳粒成型压榨,进一步盐渍,制成本发明的减少内脏脂肪用高达奶酪(Gouda cheese)型低脂肪硬质天然奶酪。
实施例11
(发酵乳的制造)
使用瑞士乳杆菌(Lactobacillus helveticus)SBT2171(FERMBP-5445)进行发酵乳的制造。将瑞士乳杆菌(Lactobacillus helveticus)SBT2171(FERM BP-5445)使用脱脂乳100g在37℃下培养12小时后的产物在新的相同培养基3kg中接种,37℃下培养12小时。将培养结束后的乳全部作为发酵剂使用,使脱脂乳100kg在32℃下发酵20小时,从而得到本发明的减少内脏脂肪用发酵乳。瑞士乳杆菌(Lactobacillushelveticus)SBT2171(FERM BP-5445)的发酵乳中的活菌数为8.2×108cells/g。
实施例12
(饮料型酸乳(Drinking yoghurt)的制造)
在实施例11中得到的发酵乳43kg中加入精制白砂糖4kg、水3kg、果胶0.15kg后进行均质化,得到50kg饮料型酸乳(Drinking yoghurt)。此饮料型酸乳(Drinking yoghurt)具有清淡、受欢迎的风味,pH为3.6,瑞士乳杆菌(Lactobacillus helveticus)SBT2171(FERM BP-5445)在饮料型酸乳(Drinking yoghurt)中的活菌数为4.6×108cells/g。
实施例13
在与实施例11同样得到的发酵物5kg中添加等量的水后,用连续离心分离机进行3,500×G、20分钟的离心分离,只分离回收菌体。为去除沉淀物中含有的非菌体成分,在添加1kg水后,再次进行离心分离,重复3次此操作。最终回收瑞士乳杆菌(Lactobacillus helveticus)SBT2171(FERM BP-5445)的菌体20g。
实施例14
按照表3中所示的组成混合原料,制成本发明的减少内脏脂肪用饲料。
表3
[试验例1]
(减少内脏脂肪作用的确认)
使用各种乳酸菌,确认减少内脏脂肪作用。
将小鼠脂肪细胞3T3-L1株在含有10%胎牛血清的Dulbecco改良Eagle培养基中培养到融合后,用胰蛋白酶处理将细胞从塑料壁面上剥下,回收于离心分离管中。通过离心分离(1,000×G,10分钟)回收细胞后,用含有0.9重量%食盐的磷酸缓冲液(PBS、pH7.2)分散为2×107个/ml的浓度。在Balb/c系3周龄的56只裸鼠的肠间膜脂肪组织内,按照每次1×107个(0.5ml)移植3T3-L1细胞。用一般标准饲料CE2饲养4星期后,使用实验动物用微型CT装置每隔1星期测定内脏脂肪量,确认内脏脂肪为充分积累。而且,用将各种乳酸菌以0.1%含量配合的CE2饲料饲养4星期后,通过解剖后计量内脏脂肪量来进行评价。动物实验为1组4只。
结果如表4所示。相对于未摄取乳酸菌的小鼠及摄取嗜热链球菌(Streptococcus thermophilis)的小鼠的内脏脂肪量,显示出摄取配合了乳杆菌属乳酸菌及乳球菌属乳酸菌的饲料的小鼠的内脏脂肪量为显著的低值。
表4
平均±标准偏差(∩=4)
Claims (3)
1.格氏乳杆菌SBT2055株、格氏乳杆菌JCM1131株、格氏乳杆菌ATCC19992株、瑞士乳杆菌SBT2171株、瑞士乳杆菌ATCC10386株、瑞士乳杆菌ATCC10797株、乳酸乳球菌JCM1107株、乳脂乳球菌ATCC11602株、乳脂乳球菌ATCC14365株、乳脂乳球菌ATCC19257株中的任意一种以上的菌体及/或其培养物在制备内脏脂肪减少剂中的用途。
2.根据权利要求1所述的用途,其特征在于,所述内脏脂肪减少剂为减少内脏脂肪用片剂、散剂、胶囊剂、饮食品或饲料。
3.根据权利要求2所述的用途,其特征在于,所述饮食品为发酵乳、健康食品或奶酪类。
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