CN112385846B - 凝结芽孢杆菌用于制备解酒的组合物的用途及食用产品 - Google Patents
凝结芽孢杆菌用于制备解酒的组合物的用途及食用产品 Download PDFInfo
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- CN112385846B CN112385846B CN202010820593.1A CN202010820593A CN112385846B CN 112385846 B CN112385846 B CN 112385846B CN 202010820593 A CN202010820593 A CN 202010820593A CN 112385846 B CN112385846 B CN 112385846B
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Abstract
本发明公开一凝结芽孢杆菌TCI711用于制备解酒的组合物的用途及食用产品,且此凝结芽孢杆菌TCI711寄存于德国微生物保藏中心,寄存编号为DSM33163。
Description
技术领域
本发明关于一种凝结芽孢杆菌,特别是关于一种凝结芽孢杆菌TCI711(Bacilluscoagulans TCI711)用于制备解酒的组合物的用途以及含凝结芽孢菌TCI711的用以解酒的食品产品。
背景技术
现代人不论是为了社交或是个人喜好,常有饮酒的情况发生。人体在饮酒之后,大部分的酒精(alcohol)会在前半段的肠道被吸收,所吸收的酒精被引导到肝脏并代谢成乙醛(acetaldehyde)。若是数量不多的乙醛,大多可以顺利代谢成乙酸(Acetic acid),最后就会变成二氧化碳跟水排出体外。
但是过多的乙醛若是代谢不及,则会对人体造成各种无法回复的损害。其中,乙醛是公认的一级致癌物质,并且饮酒之后90%以上的酒精代谢产物都要经由肝脏进行处理,也因此过度饮酒常造成肝脏的各种损伤。
发明内容
在中国台湾专利公开号TWI669124B中,发明人已公开一种凝结芽孢杆菌TCI711用于制备代谢重金属组成物的用途。并且提供实验证实在一般情况下凝结芽孢杆菌TCI711可以提升肝细胞抗氧化能力,凝结芽孢杆菌TCI711亦可提升肝细胞的线粒体活性,以及凝结芽孢杆菌TCI711具有降低脂肪肝形成的机率。但是,为了提升凝结芽孢杆菌TCI711的价值,发明人继续研究开发凝结芽孢杆菌TCI711或其相关产品的其他用途。
有鉴于此,本发明提供一种凝结芽孢杆菌TCI711(Bacillus coagulans TCI711)用于制备解酒的组合物的用途以及一种用以解酒的食品、保健食品或膳食补充品。
在一些实施例中,一种凝结芽孢杆菌TCI711(Bacillus coagulans TCI711)用于制备解酒的组合物的用途,其中凝结芽孢杆菌TCI711寄存于德国微生物保藏中心(寄存编号为DSM33163)。
在一些实施例中,凝结芽孢杆菌TCI711包含多种酵素或蛋白质,其中包括比例大于1%的酒精去除酶(alcohol dehydrogenase)及比例大于4%的醛去氢酶(aldehydedehydrogenase)。
在一些实施例中,凝结芽孢杆菌是经由于肠壁形成保护膜来减少酒精吸收以达到解酒的功能。
在一些实施例中,凝结芽孢杆菌是经由提升肝细胞线粒体活性来达到解酒的功能。线粒体为肝细胞的各种代谢活动提化学能量,当肝细胞内的线粒体活性提升时,肝细胞的对于酒精的代谢功能也能一并提升。
在一些实施例中,凝结芽孢杆菌具备酸碱耐受性,其中酸碱耐受性范围为pH值3到pH值7。
在一些实施例中,凝结芽孢杆菌的肠道定殖率为每个肠道细胞附着346CFU。
在一些实施例中,凝结芽孢杆菌的有效剂量为1x1010cells/天。
在一些实施例中,一种用以解酒的食品产品,其中包含有效剂量的凝结芽孢杆菌TCI711(Bacillus coagulans TCI711),凝结芽孢杆菌TCI711的寄存编号为DSM33163。
在一些实施例中,上述食品产品中的凝结芽孢杆菌TCI711有效剂量为1x1010cells/天。
综上所述,任一实施例的凝结芽孢杆菌TCI711或其代谢产品可制备酒解用途的组合物。换言之,前述的组合物具有下列一种或多种功能:解酒保肝、减少酒精的吸收、提升肝细胞的代谢能力。任一实施例的凝结芽孢杆菌藉由酒精去除酶及醛去除酶直接代谢被凝结芽孢杆菌所接触到的酒精。任一实施例的凝结芽孢杆菌藉其自身对于酸碱的耐受性高,可以顺利通过胃酸达到肠道。任一实施例的凝结芽孢杆菌藉由高定殖率可以在肠道壁形成保护膜以减少小肠对酒精的吸收量。任一实施例的凝结芽孢杆菌只要每日食用1x1010 cells即能有效提升酒精在体内的代谢速率。
以下结合附图和具体实施例对本发明进行详细描述,但不作为对本发明的限定。
附图说明
图1是模拟胃肠道消化实验结果图。
图2是人类结肠细胞生长状态图。
图3是凝结芽孢杆菌TCI711定殖于人类结肠细胞状态图。
图4是图3中A部分的局部放大图。
图5是凝结芽孢杆菌TCI711酒精代谢实验结果图。
图6是凝结芽孢杆菌TCI711成分及含量分析结果图。
图7是凝结芽孢杆菌TCI711人体酒精代谢实验结果图。
其中,附图标记:
BC:凝结芽孢杆菌TCI711
C2B:人类结肠细胞
A:局部放大范围
具体实施方式
在一实施例中,凝结芽孢杆菌TCI711(Bacillus coagulans TCI711)是一种可以产生乳酸的革兰式阳性菌,可于厌氧环境下生长,为兼性厌氧乳酸菌。凝结芽孢杆菌在不良的生长环境(例如:超过50℃)下会产生内孢子而停止生长,上述孢子被食入后经与胃酸反应再进到小肠时,在小肠内会从孢子状态回复而继续生长与繁殖,因此凝结芽孢杆菌TCI711在孢子状态下具有耐酸与耐热的特性。
在一些实施例中,凝结芽孢杆菌TCI711可用于制备解酒的组合物。并且,另一些实施例中,含有凝结芽孢杆菌TCI711的食品产品具备有解酒的用途。
在一些实施例中,凝结芽孢杆菌TCI711可直接代谢酒精。
在一些实施例中,凝结芽孢杆菌TCI711包含多种酵素或蛋白质,例如:50S核糖体蛋白质(50S ribosomal protein)、小酸溶性孢子蛋白(small,acid-soluble sporeprotein)、30S核糖体蛋白质(30S ribosomal protein)、延长因子tu(Elongation factorTu)三磷酸甘油醛(Glyceraldehyde-3-phosphate)醛去氢酶(Aldehyde dehydrogenase)、未定义蛋白(Uncharacterized protein)、鸟氨酸氨基转移酶(Ornithineaminotransferase)、核糖休眠促进因子(Ribosome hibernation promoting factor)、短链去氢酶(Short-chain dehydrogenase)、Pfpl家族细胞内蛋白酶(Pfpl familyintracellular protease)、琥珀酰辅酶A合成酶(Succinate CoA ligase)、二氢脂酰胺乙酰转移酶(Dihydrolipoamide acetyltransferase)、3-氧酰基-[酰基载体蛋白]还原酶还原酶(3-oxoacyl-[acyl-carrier protein]reductase)、UPF0180蛋白(UPF0180 proteinHMPREF3212_01356)、晚期胚胎发育多样化蛋白(Late embryogeneis abundant protein)、葡萄糖激酶(Glucokinase)、果糖双磷酸酶(Fructose-1,6-bisphosphatase)、二氢脂酰赖氨酸残基(Dihydrolipoyllysine-residue)、冷休眠蛋白CspB(Cold shock proteinCspB)、醛缩酶(Aldolase 2)、酒精去除酶(alcohol dehydrogenase)。其中,包括相对于凝结芽孢杆菌TCI711总体的蛋白质含量比例大于1%的酒精去除酶(alcoholdehydrogenase)及相对于凝结芽孢杆菌TCI711总体的蛋白质含量比例大于4%的醛去氢酶(aldehyde dehydrogenase)。
在一些实施例中,凝结芽孢杆菌TCI711具有耐胃酸胆盐的功能。举例来说,凝结芽孢杆菌TCI711于胃模拟环境(pH值3)的存活率为70%,且于肠道模拟环境(pH值7)的存活率为90%。
在一些实施例中,凝结芽孢杆菌TCI711可在人体肠胃道环境定殖生长,进而在肠壁形成保护膜状态。藉此,凝结芽孢杆菌TCI711可有效减少肠壁对酒精的吸收率,减少饮酒对于人体的伤害。
在一些实施例中,凝结芽孢杆菌TCI711有效剂量系1x1010 cells/天。
在一些实施例中,前述的组合物包含特定含量的凝结芽孢杆菌TCI711。
在一些实施例中,前述的组合物可为医药品。换言之,此医药品包含有效剂量的凝结芽孢杆菌TCI711。
在一些实施例中,前述的医药品可利用熟习此技艺者所详知的技术而被制造成适合于经肠地道、非经肠地道(parenterally)、口服的、或局部地(topically)投药剂型。
在一些实施例中,经肠道或口服的投药剂型可为,但不限于,锭剂(tablet)、片剂(troche)、口含锭(lozenge)、丸剂(pill)、胶囊(capsule)、分散性粉末(dispersiblepowder)或细颗粒(granule)、溶液、悬浮液(suspension)、乳剂(emulsion)、糖浆(syrup)、酏剂(elixir)、浓浆(slurry)或类似的物。在一些实施例中,非经肠地道或局部地投药剂型可为,但不限于,注射品(injection)、无菌的粉末(sterile powder)、外部制剂(externalpreparation)或类似之物。在一些实施例中,注射品的投药方式可为皮下注射(subcutaneous injection)、表皮内注射(intraepidermal injection)、皮内注射(intradermal injection)或病灶内注射(intralesional injection)。
在一些实施例中,前述的医药品可包含被广泛地使用于药物制造技术的医药上可接受的载剂(pharmaceutically acceptable carrier)。在一些实施例中,医药上可接受的载剂可为下列载剂中一种或多种:溶剂(solvent)、缓冲液(buffer)、乳化剂(emulsifier)、悬浮剂(suspending agent)、分解剂(decomposer)、崩解剂(disintegrating agent)、分散剂(dispersing agent)、黏结剂(binding agent)、赋形剂(excipient)、安定剂(stabilizing agent)、螯合剂(chelating agent)、稀释剂(diluent)、胶凝剂(gellingagent)、防腐剂(preservative)、润湿剂(wetting agent)、润滑剂(lubricant)、吸收延迟剂(absorption delaying agent)、脂质体(liposome)以及类似之物。关于选用的载剂的种类与数量是落在熟习此项技术的人士的专业素养与例行技术范畴内。在一些实施例中,作为医药上可接受的载剂的溶剂可为水、生理盐水(normal saline)、磷酸盐缓冲液(phosphate buffered saline,PBS)、或含有醇的水性溶液(aqueous solutioncontaining alcohol)。
在一些实施例中,前述的食品产品包含特定含量的凝结芽孢杆菌TCI711。
在一些实施例中,食品产品可为一般食品、保健食品或膳食补充品。换言之,此一般食品、保健食品(health foods)或膳食补充品包含有效剂量的凝结芽孢杆菌TCI711。
在一些实施例中,前述的食品产品可利用熟习此技艺者所详知的技术而被制造成适合于口服的剂型。在一些实施例中,前述的一般食品可为食品产品本身或为另一食品产品的添加物(food additive)。在一些实施例中,一般食品可为但不限于:饮料(beverages)、发酵食品(fermented foods)、烘培产品(bakery products)或调味料。
实验一:菌株的活化
首先,将保存在甘油的凝结芽孢杆菌TCI711(BCRC910807)接种于MRS培养基(BDDifcoTMLactobacilli MRS Broth,1%(v/v))中,并于37℃且厌氧环境下培养过夜并确认单一菌落形成,以得到第一次活化的凝结芽孢杆菌TCI71。
再挑取适量的第一次活化的凝结芽孢杆菌TCI711的菌落培养至15mL MRS培养基中,并于37℃且厌氧环境下培养隔夜,以形成含有第二次活化的凝结芽孢杆菌TCI711的菌液(以下简称二次活化的菌液)。
实验二:模拟胃肠道消化实验
将凝结芽孢杆菌TCI711于模拟胃酸(pH 3)(实验组A)、模拟胆汁(pH7)(实验组B)及缓冲液(pH 7)(控制组)三种待测溶液中进行测试,以确认凝结芽孢杆菌TCI711的于生物体消化道的酸碱耐受性。
实验组A采用的待测溶液为0.2穆尔浓度的氯化钾缓冲液(0.2M KCl/HCl buffer)且pH值为3。实验组B采用的待测溶液为含有0.3重量%的胆盐(购自DifcoTMOxgall,型号212820)的0.2穆尔浓度的氯化钾(0.2M KCl/HCl buffer)且pH值为7。控制组采用的待测溶液为0.2穆尔浓度氯化钾(0.2M KCl/HCl buffer)且pH值为7。
以实验一所制得的二次活化的菌液与MRS培养基配置OD600=1OD(1OD=5×108CFU)的含凝结芽孢杆菌TCI711的菌液(以下称1OD菌液)。其中,OD600为以酵素免疫分析测读仪(ELISA Reader)于600nm的波长下所测定到的吸光值(OD值)。
取100μL的1OD菌液接种至9.9毫升(mL待测溶液中。接着,使菌液与待测溶液充分混合,并在37℃且厌氧环境下培养3小时以形成待测菌液。将待测菌液取1毫升(mL)做序列倍数稀释(例如:107~108倍),再将稀释后的待测菌液以MRS培养基于37℃且厌氧环境下培养48小时,然后以平板计数法(plate count)计算各组中的凝结芽孢杆菌TCI711菌数。
请参阅图1。图式中凝结芽孢杆菌TCI711的存活力为计数后的活菌数,并以logCFU/mL表示。其中,log CFU/mL代表每毫升菌液含有的菌落形成单位(CFU,colony-formingunit)并以对数(log)表示。由图1可知,控制组的凝结芽孢杆菌TCI711的存活力为9.88logCFU/mL、实验组A的凝结芽孢杆菌TCI711的存活力为7.48log CFU/mL,以及实验组B的凝结芽孢杆菌TCI711的存活力为9.58log CFU/mL。
由此可知,相对于控制组,凝结芽孢杆菌TCI711的存活率为70%以上,换言之,凝结芽孢杆菌TCI711可于胃模拟环境(pH值3-4)存活。相对于控制组,凝结芽孢杆菌TCI711的存活率为90%以上,换言之,凝结芽孢杆菌TCI711亦可于肠道模拟环境(pH值7)存活。因此,凝结芽孢杆菌TCI711具有耐胃酸胆盐的功能。
实验三:肠道定殖实验
于此,采用人类结肠细胞C2BBel(CRL-2102TM)与凝结芽孢杆菌TCI711共同培养后,以显微镜观察其定殖状态并且以平板计数法分析定殖率,以确认凝结芽孢杆菌TCI711的肠道定殖状。肠道是人体最大的消化吸收器官,益生菌若有较高的肠道定殖率,则能够更高效的发挥其效能。
首先,取得六孔培养盘,将人类结肠细胞以每孔7.5×105个细胞的数量殖入孔中,并且每孔添加2毫升(mL)的培养液,置于5%二氧化碳浓度、温度37℃的恒温箱中培养24小时。于此,培养液是以DMEM培养基(Dulbecco's Modified Eagle Medium,Gibco,Cat.12100-038)添加10%的胎牛血清(Gibco,Cat.10438-026)、1%的青霉素/链霉素(Gibco,Cat.15140-122)和0.01mg/ml的转铁蛋白所制得。
接着,以实验一所制得的二次活化的菌液与MRS培养基配置OD600=1OD(1OD=5×108CFU)的含凝结芽孢杆菌TCI711的菌液(以下称1OD菌液)。其中,OD600为以酵素免疫分析测读仪(ELISA Reader)于600nm的波长下所测定到的吸光值(OD值)。将1OD菌液离心以搜集凝结芽孢杆菌TCI711的菌体。然后,使用不含抗生素的C2BBe1培养基将搜集到的菌体调整成108CFU/ml的实验用菌液。
将培养盘中的培养液移除后,以1×PBS(购自Gibco)清洗。于清洗后,对于实验组01与实验组02,每孔加入1毫升(mL)的实验用菌液,而对于控制组01与控制组02,则每孔加入1mL的C2BBe1培养基(不含抗生素及实验用菌液)。然后,所有组别同时在低氧环境(氧含量1%以下)下培养1小时。
于培养1小时后,将培养盘中的上清液去除后,使用2mL的1×PBS清洗五次。
然后,对实验组01与控制组01的人类结肠细胞及凝结芽孢杆菌TCI711进行革兰氏染色(染色试剂购自BaCO Biotech)后,将实验组01与控制组01置于显微镜下观察其细胞及菌体定殖情形,如图2、图3及图4。对于实验组02与控制组02,则每孔添加1毫升(mL)的Triton X-100(非离子型界面活性剂)并于室温下反应10分钟以将人类结肠细胞及凝结芽孢杆菌TCI711自培养盘中取下,然后使用稀释涂抹法于琼脂胶平板进行涂盘,并以平板计数法分析定殖率。
参考图2、图3及图4。相较于控制组01,实验组01的照片可看到在人类结肠细胞(即图4中椭圆形微凸起状阴影C2B)的外围有许多凝结芽孢杆菌TCI711(即图4中的细小且深色的细长阴影BC),此即为凝结芽孢杆菌TCI711定殖于人类结肠细胞上的形态。并且,透过涂盘计数可得到凝结芽孢杆菌TCI711的定殖率为346CFU/cells。可知,凝结芽孢杆菌TCI711能够稳定的定殖在人类结肠细胞外围。
实验四:酒精代谢实验
于此,将活化后的凝结芽孢杆菌TCI711接种于含酒精的MRS培养基一段时间后,再量测酒精含量的变化,以确认凝结芽孢杆菌TCI711是否具直接备分解酒精的能力。
首先,以实验一所制得的二次活化的菌液与MRS培养基配置OD600=1OD(1OD=5×108CFU)的含凝结芽孢杆菌TCI711的菌液(以下称1OD菌液)。其中,OD600为以酵素免疫分析测读仪(ELISA Reader)于600nm的波长下所测定到的吸光值(OD值)。
实验组是1%(v/v)的1OD菌液接种至含有5%酒精的MRS培养基中,而控制组则是直接使用相同总量的含5%酒精的MRS培养基(不接种凝结芽孢杆菌TCI711)。然后,将控制组与实验组置于同样的环境温度37℃下培育8小时。于此,MRS培养基购自BD DifcoTM。
接下来,将各组别培养后所形成的菌液收集至离心管中,以进行离心。于离心后,收集上清液。然后,将收集到的上清液在温度60℃~80℃之间下进行蒸馏1小时并收集其冷凝液。最后,以刻度式酒精度计(型号AL80)进行量测冷凝液的酒精度(W/W%)。
参照图4。控制组所量测到的酒精度为3.9%(w/w),而实验组的酒精度降为2.8%以下。换言之,相较于控制组,凝结芽孢杆菌TCI711可以在8小时内减少28%的酒精含量。基此,凝结芽孢杆菌TCI711具备直接代谢酒精的效果。
实验五:菌体蛋白质分析
首先,以实验一所制得的二次活化的菌液与MRS培养基配置OD600=0.1OD的初始菌液,并将初始菌液于37℃且厌氧环境下培养隔夜,以得到OD600=8OD的待测菌液。
将待测菌液以高速离心机(Heraeus Megafuge 16centrifuge,ThermoScientific)以5000XP转速离心20分钟,然后去除上清液,并保留沉淀物(即到菌体)。接着,以50毫升(mL)裂解缓冲液(Lysis Buffer)回溶菌体,再加入1μg/ml的氧核糖核酸酶(DNAse)并于室温下反应10分钟,以形成待破菌液。于此,裂解缓冲液是由50mM NaH2PO4、300mM NaCl及1mM MgCl2所制得,并且其pH值为7。
反应后,将待破菌液以高压破菌机(Constant System TS series CF1)分别以25Kpsi、30Kpsi及32Kpsi三种压力进行三次破菌以得到破菌液。
将破菌液以冷冻抽干机(EYELA)在-80℃低温及真空环境下干燥24小时,把破菌液中的水分直接以升华为水蒸气的方式除去以得到干燥后的破菌。
接着,以乙腈及三氟乙酸为溶剂将上述干燥后的破菌配置为30mg/ml的样本,将样本使用高效液相色谱仪(High Performance Liqid Chromatography)设定检测波长220,280nm,流速0.5ml/min,柱温40℃,进样体积20μl,梯度设定ACN0%-45%进行分离,并将分离出的胜肽分馏物(fractions)以冷冻抽干机(EYELA)在-80℃低温及真空环境下干燥24小时得粗分离胜肽。
将粗分离胜肽以300μ的无菌水回溶后,取出10μl粗分离胜肽水溶液以奈米级液相层析仪(UltiMate 3000RSLCnano LC Systems)进行分离,再经由飞行时间式串联质谱仪系统(Q-TOF Mass Spectrometry:6600System)分析所分离出来的胜肽的分子质量,并将质量比对于NCBI及UniProt数据库,可得到结果包括:50S核糖体蛋白质(50Sribosomal protein)、小酸溶性孢子蛋白(small,acid-soluble spore protein)、30S核糖体蛋白质(30S ribosomal protein)、延长因子tu(Elongation factor Tu)、三磷酸甘油醛(Glyceraldehyde-3-phosphate)、醛去氢酶(Aldehyde dehydrogenase)、未定义蛋白(Uncharacterized protein)、鸟氨酸氨基转移酶(Ornithine aminotransferase)、核糖休眠促进因子(Ribosome hibernation promoting factor)、短链去氢酶(Short-chaindehydrogenase)、Pfpl家族细胞内蛋白酶(Pfpl family intracellular protease)、琥珀酰辅酶A合成酶(Succinate CoA ligase)、二氢脂酰胺乙酰转移酶(Dihydrolipoamideacetyltransferase)、3-氧酰基-[酰基载体蛋白]还原酶还原酶(3-oxoacyl-[acyl-carrier protein]reductase)、UPF0180蛋白(UPF0180 protein HMPREF3212_01356)、晚期胚胎发育多样化蛋白(Late embryogeneis abundant protein)、葡萄糖激酶(Glucokinase)、果糖双磷酸酶(Fructose-1,6-bisphosphatase)、二氢脂酰赖氨酸残基(Dihydrolipoyllysine-residue)、冷休眠蛋白CspB(Cold shock protein CspB)、醛缩酶(Aldolase 2)、及酒精去除酶(alcohol dehydrogenase),如图5所示。
参考图5。在凝结芽孢杆菌TCI711中,含量最高的为50S核糖体蛋白质,50S核糖体蛋白质的含量相对于凝结芽孢杆菌TCI711总体蛋白质含量的比例为14.8%。含量次高的是小酸溶性孢子蛋白,小酸溶性孢子蛋白的含量相对于凝结芽孢杆菌TCI711总体蛋白质含量的比例为9.47%。其余蛋白质中30S核糖体蛋白质的含量为8.64%、延长因子tu的含量为7.82%、三磷酸甘油醛的含量为5.35%、未定义蛋白的含量为4.12%、鸟氨酸氨基转移酶的含量为3.7%、核糖休眠促进因子的含量为3.29%、短链去氢酶(Short-chaindehydrogenase)的含量为%、Pfpl家族细胞内蛋白酶的含量为2.88%、琥珀酰辅酶A合成酶的含量为2.47%、二氢脂酰胺乙酰转移酶和3-氧酰基-[酰基载体蛋白]还原酶还原酶的含量为1.65%。其中,UPF0180蛋白、晚期胚胎发育多样化蛋白、葡萄糖激酶、果糖双磷酸酶、二氢脂酰赖氨酸残基、冷休眠蛋白CspB及醛缩酶的含量为1.23%。
并且,凝结芽孢杆菌TCI711还包括相对于凝结芽孢杆菌TCI711总体蛋白质含量的比例大于1.23%的酒精去除酶(alcohol dehydrogenase)及相对于凝结芽孢杆菌TCI711总体蛋白质含量的比例为4.53%的醛去氢酶(aldehyde dehydrogenase)。其中,酒精去除酶用以将酒精转化为乙醛,而醛去氢酶用以将乙醛转化为乙酸。
由此结果可知,凝结芽孢杆菌TCI711具有代谢酒精相关蛋白质,以致能够在肠道中直接代谢酒精。
实验六:人体试验
于此,进行人体试验以确认服用凝结芽孢杆菌TCI711实际上对于人体解酒效能的影响。试验分为控制组及实验组,每组别各5位受试者。
控制组:受试者未曾服用过凝结芽孢杆菌TCI711并于饭后的30钟内喝完75毫升(mL)且酒精浓度40%的可饮用酿造酒(Jack Daniel’s Bourbon Whisky),然后以酒精呼气仪(LION Alcometer 400)分别在饮酒后0分钟(即喝完马上测试)、30分钟、60分钟、90分钟及120分钟共5个检测点量测各受试者的酒精值。
实验组:受试者于每日饭后服用含凝结芽孢杆菌TCI711胶囊(即每天服用1X1010cells的凝结芽孢杆菌TCI711)并连续服用1周,再于最后一日胶囊服用后的30钟内喝完75毫升(mL)且酒精浓度40%的可饮用酿造酒,然后以酒精呼气仪(LION Alcometer 400)分别在前述的5个检测点量测各受试者的酒精值。于此,胶囊中的凝结芽孢杆菌TCI711是取自于实验一所制得的二次活化的菌液。
于此,将同组别且同检测点的5位受试者的酒精值取平均,以得到测试结果(如下表一及图5所示)。
表一
0分钟 | 30分钟 | 60分钟 | 90分钟 | 120分钟 | |
控制组(mg/L) | 0.192 | 0.092 | 0.062 | 0.048 | 0.026 |
实验组(mg/L) | 0.118 | 0.030 | 0.024 | 0.006 | 0.000 |
参照表一及图5,在饮酒后0分钟,实验组的酒精度明显低于控制组;由此可得知服用凝结芽孢杆菌TCI711的受试者对于酒精的吸收度下降。也就是说,持续服用一周凝结芽孢杆菌TCI711的受试者已经在肠壁形成保护层,并具备直接分解并减少吸收酒精的效能。
在饮酒后90分钟,实验组的酒精值已趋近0,即在饮酒后的90分钟内,凝结芽孢杆菌TCI711几乎快要将受试者体内的酒精分解完。在饮酒后120分钟,实验组的酒精值已为0mg/L,意即在饮酒后的2个小时内,凝结芽孢杆菌TCI711已经将受试者体内的酒精完全分解。也就是说,持续服用一周的凝结芽孢杆菌TCI711的受试者相对于未服用过凝结芽孢杆菌TCI711的受试者能够更快速地代谢体内的酒精。
虽然本发明的技术内容已经以较佳实施例揭露如上,然其并非用以限定本发明,任何熟习此技艺者,在不脱离本发明的精神所作些许的更动与润饰,皆应涵盖于本发明的范畴内,因此本发明的保护范围当视后附的权利要求所界定者为准。
当然,本发明还可有其它多种实施例,在不背离本发明精神及其实质的情况下,熟悉本领域的技术人员可根据本发明作出各种相应的改变和变形,但这些相应的改变和变形都应属于本发明权利要求的保护范围。
【生物材料寄存】
德国微生物保藏中心(德国);2019年5月24日;寄存编号:DSM33163。
Claims (6)
1.凝结芽孢杆菌在制备于肠壁形成保护膜来减少酒精吸收以达到解酒的组合物中的用途,其特征在于,所述凝结芽孢杆菌为Bacillus coagulans TCI711,寄存编号DSM33163,所述凝结芽孢杆菌经由稳定的定殖在人类结肠细胞外围于所述肠壁形成所述保护膜,所述凝结芽孢杆菌具备酸碱耐受性,其中所述酸碱耐受性范围为pH值3到pH值7,所述凝结芽孢杆菌的有效剂量为1x1010 cells/天。
2.根据权利要求1所述的用途,其特征在于,所述凝结芽孢杆菌是经由直接代谢酒精以达到所述解酒的功能。
3.根据权利要求2所述的用途,其特征在于,所述凝结芽孢杆菌包含酒精去除酶及醛去氢酶。
4.根据权利要求2所述的用途,其特征在于,所述凝结芽孢杆菌是经由提升肝细胞线粒体活性来达到所述解酒的功能。
5.根据权利要求2所述的用途,其特征在于,所述凝结芽孢杆菌的肠道定殖率为每个肠道细胞附着346 CFU。
6.一种组合物,其用以于肠壁形成保护膜来减少酒精吸收以达到解酒,其特征在于,其包含有效剂量的凝结芽孢杆菌TCI711,所述凝结芽孢杆菌TCI711的寄存编号为DSM33163,所述凝结芽孢杆菌经由稳定的定殖在人类结肠细胞外围于所述肠壁形成所述保护膜,所述凝结芽孢杆菌具备酸碱耐受性,其中所述酸碱耐受性范围为pH值3到pH值7,所述凝结芽孢杆菌的有效剂量为1x1010 cells/天。
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