CN102482662A - 硫化氢生成酶抑制剂 - Google Patents
硫化氢生成酶抑制剂 Download PDFInfo
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- CN102482662A CN102482662A CN2010800397020A CN201080039702A CN102482662A CN 102482662 A CN102482662 A CN 102482662A CN 2010800397020 A CN2010800397020 A CN 2010800397020A CN 201080039702 A CN201080039702 A CN 201080039702A CN 102482662 A CN102482662 A CN 102482662A
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- Prior art keywords
- hydrogen sulfide
- catechin
- enzyme
- inhibition
- generates
- Prior art date
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Classifications
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- C—CHEMISTRY; METALLURGY
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/99—Enzyme inactivation by chemical treatment
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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
- 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
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- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
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Abstract
本发明的目的是抑制硫化氢产生,特别是提供一种硫化氢生成酶抑制剂。本发明人经认真研究后发现,儿茶素类具有硫化氢生成酶抑制活性,从而完成了本发明。即,本发明提供一种以儿茶素类作为有效成分的硫化氢生成酶抑制剂。
Description
技术领域
本发明涉及一种以儿茶素类作为有效成分的硫化氢生成酶抑制剂。
背景技术
硫化氢是具有刺激性气味的气体,已知具有刺激皮肤粘膜和引起呼吸麻痹等毒性。其毒性在社会学领域也被视作问题,希望除去硫化氢。在自然界中,硫化氢包含于火山气体和温泉等中,在人为方面,硫化氢通过石油化学工业等产生。此外,硫化合物有时被厌氧性细菌等还原而产生硫化氢,例如厌氧性细菌有时在污物和废水等中繁殖而产生硫化氢。此外,产生硫化氢的细菌中,也有作为口腔内细菌或肠内细菌在机体内生存的细菌。这些细菌以食物残渣和机体内的蛋白质为原料产生硫化氢。还已知机体内生成内源性硫化氢。
已知硫化氢的毒性机理是因硫化氢的高反应性而导致对皮肤粘膜的刺激以及细胞色素c氧化酶的抑制。细胞色素c氧化酶抑制作用迅速发生,因此暴露于高浓度的硫化氢时,肺的氧分压急剧降低,以至昏倒。此外,认为硫化氢在脑内增强NMDA受体的活性(非专利文献1)。
如上所述,硫化氢也由口腔内细菌或肠内细菌产生。由口腔内细菌或肠内细菌产生的硫化氢与自然界中或工业水平上产生的硫化氢相比规模较小。但是,如果机体内产生硫化氢,则该个体将以非常封闭的状态暴露于硫化氢,其影响是直接的。此外,产生的硫化氢会带来口臭、肠内气体臭之类的惹人厌恶、令人不快的气味。因此,对于由口腔内细菌或肠内细菌产生的硫化氢,希望对其进行阻断或抑制。另外,呼气中的挥发性硫化物(VSC)中,硫化氢、甲硫醇和二甲硫醚约占90%,硫化氢可以说是口臭的主要原因之一。
在口腔内,口腔内的脱落上皮细胞和白细胞、食物残渣等被来源于细菌的蛋白酶分解成半胱氨酸和甲硫氨酸等含硫氨基酸,然后在硫化氢生成酶和甲硫氨酸酶的作用下生成硫化氢。作为口腔内的硫化氢生成酶,报道了半胱氨酸脱硫基酶和胱硫醚β合成酶等。
专利文献1揭示了由假单胞菌属细菌和大肠杆菌纯化的挥发性硫化合物生成抑制剂。但是,该抑制剂的目的是应用于以特定的醛、十一烯醛、脂环族酮等香料作为有效成分的家用制品,无法安全地应用于人体等。
专利文献2揭示了含有水溶性难消化性糖类作为有效成分的减少肠内硫化氢的组合物。但是,专利文献2的减少肠内硫化氢的效果仅限于肠内,其减少机理也不明。
专利文献3揭示了阻断、抑制机体内的硫化氢的生成的镇痛用组合物。但是,专利文献3的包含DL-炔丙基甘氨酸和β-氰基苯胺的镇痛用组合物是抑制胱硫醚γ裂合酶、抑制机体内的内源性硫化氢的生成的组合物。
非专利文献4揭示了儿茶素衍生物抑制来源于牙龈卟啉单胞菌(Porphyromonas gingivalis)的蛋白酶。但是,关于硫化氢生成酶的抑制既没有记载也没有启示。
非专利文献5揭示了绿茶儿茶素抑制金属蛋白酶。
但是,关于硫化氢生成酶的抑制既没有记载也没有启示。
现有技术文献
专利文献
专利文献1:日本专利特开2008-173441号公报
专利文献2:日本专利特开2007-99672号公报
专利文献3:日本专利特开2007-112735号公报
非专利文献
非专利文献1:Abe,K,Kimura H,J.神经科学(Neurosci.)16,1066-71,1996
非专利文献2:Persson S,Edlund M-B,Claesson R,Carlsson J:口腔微生物免疫(Oral Microbiol Immunol.)1990;5:195-201
非专利文献3:Tanaka M,Yamamoto Y,Kuboniwa M,Nonaka A,NishidaN,Maeda K,Kataoka K,Nagata H,Shizukuishi S:微生物传染(Microbesand Infection)2007;6:1078-1083
非专利文献4:Okamoto M,Sugimoto A,Leung K-P,Nakayama K,Kamaguchi A,Maeda N:口腔微生物免疫(Oral Microbiol.Immunol.)2004;19:118-120
非专利文献5:Demeule M,Brossard M,Page M,Gingras D,BeliveauR:生物化学生物物理学报(Biochim Biophys Acta.)2000;1478:51-60.
发明内容
发明所要解决的技术问题
本发明的目的是抑制硫化氢产生,特别是提供一种硫化氢生成酶抑制剂。
解决技术问题所采用的手段
本发明人为了解决上述课题,着眼于无副作用且安全性高的自古以来就为人所利用的来源于绿茶的成分,实施了针对细菌的硫化氢生成的抑制实验,结果发现,儿茶素类具有硫化氢生成酶抑制活性,从而完成了本发明。
发明的效果
根据本发明,通过使用儿茶素类作为有效成分,能有效地抑制硫化氢生成酶,阻断、抑制硫化氢的生成。
此外,儿茶素类是来源于绿茶的成分,安全性高,给人的感觉好。因此,以儿茶素类作为有效成分的硫化氢生成酶抑制剂容易被消费者接受。
附图的简单说明
图1A所示为由比色分析得到的、EGCg和氯化锌对半胱氨酸脱硫基酶和甲硫氨酸酶的抑制活性的结果。图1A是F.n的结果。
图1B所示为由比色分析得到的、EGCg和氯化锌对半胱氨酸脱硫基酶和甲硫氨酸酶的抑制活性的结果。图1B是P.g的结果。
图1C所示为由比色分析得到的、EGCg和氯化锌对半胱氨酸脱硫基酶和甲硫氨酸酶的抑制活性的结果。图1C是T.d的结果。
图2A所示为由气相色谱法分析得到的、EGCg和氯化锌对硫化氢生成酶和甲硫氨酸酶的抑制活性的结果。图2A是F.n的结果。
图2B所示为由气相色谱法分析得到的、EGCg和氯化锌对硫化氢生成酶和甲硫氨酸酶的抑制活性的结果。图2B是P.g的结果。
图2C所示为由气相色谱法分析得到的、EGCg和氯化锌对硫化氢生成酶和甲硫氨酸酶的抑制活性的结果。图2C是T.d的结果。
实施发明的方式
本发明提供一种以儿茶素类作为有效成分的硫化氢生成酶抑制剂。
儿茶素(catechin)狭义上是指C15H14O6、分子量290.27、在木本植物的木芯中大量含有的水溶性多酚,广义上包括作为其衍生物的多酚。本说明书中,为了使定义明确,儿茶素是指狭义的儿茶素,提及儿茶素类时,是指儿茶素以及作为其衍生物的一系列多酚。作为儿茶素类,特别是已知茶中所含的茶儿茶素。作为儿茶素类,已知(+)-儿茶素、(-)-表儿茶素、(-)-表没食子儿茶素、(-)-表儿茶素没食子酸酯、(-)-表没食子儿茶素没食子酸酯、(-)-没食子儿茶素没食子酸酯、(-)-儿茶素没食子酸酯等。
本发明还供一种以儿茶素类的(-)-表儿茶素没食子酸酯、(-)-表没食子儿茶素没食子酸酯、(-)-没食子儿茶素没食子酸酯、(-)-儿茶素没食子酸酯或它们的混合物作为有效成分的硫化氢抑制剂。
本发明还提供一种以(-)-表儿茶素没食子酸酯、(-)-表没食子儿茶素没食子酸酯、(-)-没食子儿茶素没食子酸酯、(-)-儿茶素没食子酸酯的任一种或它们的混合物作为有效成分的、抑制来源于具核梭杆菌(Fusobacterium nucleatum)、牙龈卟啉单胞菌(Porphyromonas gingivalis)或齿垢密螺旋体(Treponema denticola)的硫化氢生成酶的硫化氢生成酶抑制剂。
具核梭杆菌、牙龈卟啉单胞菌、齿垢密螺旋体均已知为口腔内细菌,还已知它们的硫化氢和甲硫醇的生成能力高,是牙周病菌(非专利文献2、非专利文献3)。
本发明还提供一种含有所述硫化氢生成酶抑制剂的饮食品。饮食品包括生鲜食品、肉、鱼等动物性食品,谷物、蔬菜等植物性食品,乳制品、面包、即食食品等加工食品,点心类等嗜好食品,甜味剂、调味剂等烹饪调味用材料,保健食品,特殊用途食品,水、清凉饮料水、酒精饮料、茶等饮料,食品加工材料,食品添加剂等。
本发明还提供一种含有所述硫化氢生成酶抑制剂的口腔用组合物。作为口腔用组合物,可以采用牙膏、液体牙膏、液状牙膏、湿牙粉等牙膏类,漱口水,口腔清洗剂,含片剂,口香糖等形态。
下面例举本发明的例子进行说明,但本发明的范围不限定于以下的例子。
实施例1
进行关于儿茶素类对硫化氢生成酶和甲硫氨酸酶的酶抑制活性的试验。即,使用半胱氨酸和甲硫氨酸作为底物,添加来源于具核梭杆菌、牙龈卟啉单胞菌、齿垢密螺旋体的粗酶液,将各试剂混合,对作为甲硫氨酸酶产物的α-氧代丁酸、甲硫醇和作为硫化氢生成酶产物的丙酮酸、硫化氢进行定量,藉此测定对甲硫氨酸酶和硫化氢生成酶的抑制活性。
材料及方法
1-1材料
1-1-1硫化氢生成酶抑制剂候选对象
使用以下试剂作为硫化氢生成酶抑制剂。即,作为儿茶素类,使用(+)-儿茶素、(-)-表儿茶素、(-)-表没食子儿茶素、(-)-表儿茶素没食子酸酯、(-)-表没食子儿茶素没食子酸酯、(-)-没食子儿茶素没食子酸酯、(-)-儿茶素没食子酸酯(儿茶素类购自长良科学公司)以及没食子酸、氯化锌。这些抑制剂候选对象在本说明书中缩写为:(+)-儿茶素:C、(-)-表儿茶素:EC、(-)-表没食子儿茶素:EGC、(-)-表儿茶素没食子酸酯:ECg、(-)-表没食子儿茶素没食子酸酯:EGCg、(-)-没食子儿茶素没食子酸酯:GCg、(-)-儿茶素没食子酸酯:Cg、没食子酸:G、氯化锌:ZnCl2。已知没食子酸具有杀菌活性,其衍生物被用作口腔用组合物。已知氯化锌具有口臭预防活性,被用于漱口水。
1-1-2使用菌株
作为硫化氢生成细菌,使用具核梭杆菌(本说明书中缩写为F.n)、牙龈卟啉单胞菌(本说明书中缩写为P.g)、齿垢密螺旋体(本说明书中缩写为T.d)。作为菌株,具核梭杆菌使用JCM8532菌株、牙龈卟啉单胞菌使用W83菌株、齿垢密螺旋体使用ATCC保藏编号35405的菌株。
1-2试验方法
1-2-1各菌的粗酶液的制备
将F.n接种于15ml胰蛋白胨大豆肉汤(Trypticase Soy Broth,TSB),进行厌氧培养,直至生长到达稳定期为止。另外,胰蛋白胨大豆肉汤(TSB)如下所述制备:将3g胰蛋白胨大豆肉汤(DIFCO公司)和0.3g酵母提取物(Yeast Extract)(DIFCO公司)溶解于100ml水后,添加0.1ml氯高铁血红素(hemin)(5mg/ml,0.1N NaOH中)和0.1ml甲萘醌(menadione)(0.5mg/ml,50%EtOH中)。
将上述培育作为预培育,再接种于200ml TSB,于37℃厌氧培养,直至生长到达稳定期后24小时(培养时间2天)为止。
接着,以3000×g、10min、4℃的条件离心分离培养液,除去培养基成分,回收菌体。然后,添加50mM Tris-HCl缓冲液(pH7.5)(将6.05g三羟甲基氨基甲烷(分子量121)溶解于水,用HCl调整至pH7.5后进行定容,调整至1l),再离心,洗涤菌体后,除去缓冲液,将菌体于-80℃冷冻。然后,将菌体悬浮于20-25ml的Tris-HCl缓冲液中,将细胞膜超声波粉碎(20W,30秒×15次,0℃)。
以20000×g、30min、4℃的条件对其进行离心,将上清作为粗酶液。粗酶液用DC蛋白定量试剂盒(DC Protein Assay kit)(Pierce公司)进行蛋白质定量,在试验体系中,按照终浓度0.3mg蛋白质/ml的条件稀释后使用。
P.g和F.n同样用TSB培养基培养。T.d用TYGVS培养基培养。培养时间为7天。关于粗酶液的制备,P.g和T.d与F.n同样进行。
TYGVS培养基的制备如下所述。即,
将下述I液~III液以及100ml灭活兔血清混合制成。
I液:将5g心浸出液肉汤(heart infusion broth)、10g胰蛋白胨(tryptone)、10g酵母提取物、10g明胶、0.5g硫酸铵、0.5g K2HPO4、0.1gMgSO4制成750ml的水溶液,调整至pH7.0后,于121℃高压釜中放置15分钟而得;
II液:
将L-半胱氨酸HCl 1g、
葡萄糖 1g、
丙酮酸钠 0.25g、
VFA液(将1.7ml乙酸、0.6ml丙酸、0.4ml正丁酸、0.1ml正戊酸、0.1ml异戊酸、0.1ml异丁酸、0.1ml DL-甲基丁酸和27.9ml蒸馏水混合)5ml、TPP液(在0.25g焦磷酸硫胺素中添加100ml蒸馏水并混合)5ml、1-N KOH 16ml
混合,再加水制成50ml的水溶液,调整至pH7.2后,过滤灭菌而得;
III液:将1g果胶溶解于100ml蒸馏水,于121℃高压釜中放置15分钟而得。
1-2-2采用比色定量的硫化氢生成酶(半胱氨酸脱硫基酶)抑制试验
作为硫化氢生成酶的一种的半胱氨酸脱硫基酶由半胱氨酸生成丙酮酸、氨和硫化氢。采用比色定量的硫化氢生成酶抑制试验中,以半胱氨酸作为底物,以由半胱氨酸脱硫基酶生成的丙酮酸的量作为指标,将各试剂对硫化氢生成酶的抑制活性进行比较。
溶液的制备
如下所述制备300mM半胱氨酸溶液、酶-辅酶混合溶液、试样溶液、3-甲基-2-苯并噻唑啉酮腙(MBTH)反应溶液。
300mM半胱氨酸溶液:将半胱氨酸用Tris-HCl缓冲液稀释至300mM。
酶-辅酶混合溶液:在临试验前用Tris-HCl缓冲液调整,使反应体系中粗酶液的蛋白质量达到0.3mg/ml、吡哆醛-5’-磷酸达到0.05mM。
试样溶液:将氯化锌或儿茶素类用Tris-HCl缓冲液稀释至各种浓度。
3-甲基-2-苯并噻唑啉酮腙(MBTH)反应溶液:将30mg MBTH溶解于50ml蒸馏水。然后,添加100ml的1M乙酸钠缓冲液(pH5.0)(将82.03g乙酸钠溶解于水,用乙酸调整至pH5.0后进行定容,调整至1l)。
丙酮酸的定量
在0.1ml 300mM半胱氨酸溶液和0.2ml试样溶液中添加0.7ml酶-辅酶混合溶液,于37℃振荡。1小时后,添加0.5ml 6%高氯酸终止反应,离心分离(3000×g、10分钟、4℃)后,回收上清。
在0.4ml上清中添加1.2ml MBTH反应溶液,于50℃反应30分钟。反应液用乙酸钠缓冲液稀释3倍,于335nm处测定吸光度。所有试验进行2次,求出其平均值。此外还制作了丙酮酸的校正曲线。
抑制率
抑制率是将加入试样溶液时的丙酮酸量设为Pi,将添加Tris-HCl缓冲液来代替抑制剂溶液时的丙酮酸量设为P0,如下所述求得。
抑制率(%)=(P0-Pi)/P0×100
1-2-3采用比色定量的甲硫氨酸酶抑制试验
作为半胱氨酸脱硫基酶的对照,测定对甲硫氨酸酶的酶抑制活性。甲硫氨酸酶以甲硫氨酸作为底物,生成甲硫醇、氨和α-氧代丁酸。采用比色定量的甲硫氨酸酶抑制试验中,以由甲硫氨酸酶生成的α-氧代丁酸的量作为指标,将各试剂对甲硫氨酸酶的抑制活性进行比较。
试剂的制备
如下所述制备300mM甲硫氨酸溶液。酶-辅酶混合溶液、试样溶液、3-甲基-2-苯并噻唑啉酮腙(MBTH)反应溶液的制备与上文中同样地进行。
300mM甲硫氨酸溶液:将甲硫氨酸用Tris-HCl缓冲液稀释至300mM。
α-氧代丁酸的定量
在0.1ml 300mM甲硫氨酸溶液和0.2ml试样溶液中添加0.7ml酶-辅酶混合溶液,于37℃振荡。1小时后,添加0.5ml 6%高氯酸终止反应,离心分离(3000×g、10分钟、4℃)后,回收上清。
在0.4ml上清中添加1.2ml MBTH反应溶液,于50℃反应30分钟。反应液用乙酸钠缓冲液稀释3倍,于335nm处测定吸光度。所有试验进行2次,求出其平均值。此外还制作了α-氧代丁酸的校正曲线。
抑制率
甲硫氨酸酶的抑制率是将加入试样溶液时的α-氧代丁酸量设为Ki,将添加Tris-HCl缓冲液来代替抑制剂溶液时的α-氧代丁酸量设为K0,如下所述求得。
抑制率(%)=(K0-Ki)/K0×100
1-2-4采用气相色谱的硫化氢生成酶抑制试验
硫化氢生成酶由半胱氨酸生成硫化氢。采用气相色谱的硫化氢生成酶抑制试验中,以半胱氨酸作为底物,以由硫化氢生成酶生成的硫化氢的量作为指标,将各试剂对硫化氢生成酶的抑制活性进行比较。
硫化氢的定量
在试管中添加0.1ml L-半胱氨酸溶液、0.2ml试样(抑制剂候选对象)溶液、0.7ml酶-辅酶混合溶液,用橡皮塞密封,在37℃水浴中边搅拌边保温。60分钟后,添加3M磷酸水溶液终止反应后,再于37℃搅拌并保温10分钟。对于F.n抽取顶空气体15μl,对于P.g抽取顶空气体100μl,对于T.d抽取顶空气体50μl,通过气相色谱法测定硫化氢量。分析条件如下所述。
柱:HP-PLOT/Q(φ0.53mm×30m)
温度:70℃(2.5min)→以30℃/min到达190℃→190℃(3.5min)
检测器:FPD
分流比:48∶1
注入量:15、50及100μl
1-2-5采用气相色谱的甲硫氨酸酶抑制试验
作为硫化氢生成酶的对照,测定对甲硫氨酸酶的酶抑制活性。甲硫氨酸酶以甲硫氨酸作为底物,生成甲硫醇、氨和α-氧代丁酸。采用气相色谱的甲硫氨酸酶抑制试验中,以由甲硫氨酸酶生成的甲硫醇的量作为指标,将各试剂对甲硫氨酸酶的抑制活性进行比较。
甲硫醇的定量
在试管中添加0.1ml L-甲硫氨酸溶液、0.2ml试样溶液、0.7ml酶-辅酶混合溶液,用橡皮塞密封,在37℃水浴中边搅拌边保温。60分钟后,添加3M磷酸水溶液终止反应后,再于37℃搅拌并保温10分钟。对于F.n抽取顶空气体40μl,对于P.g和T.d抽取顶空气体200μl,通过气相色谱法测定甲硫醇量。分析条件如上所述。
2-3结果
2-3-1采用比色定量的、儿茶素类对F.n的硫化氢生成酶(半胱氨酸脱硫基酶)和甲硫氨酸酶的抑制试验的结果
1-2-2的结果示于表1、2,1-2-3的结果示于表3。儿茶素类对F.n的硫化氢生成酶(半胱氨酸脱硫基酶)的抑制活性从高到低依次为Cg>GCg>EGCg>ECg>没食子酸、(+)-C,特别是Cg、GCg、EGCg和ECg具有高抑制活性。儿茶素类对F.n的甲硫氨酸酶的抑制活性从高到低依次为EGCg>ECg>EGC>EC、没食子酸、C,特别是EGCg和ECg具有高抑制活性。关于此时的IC50,EGCg为0.4mM,ECg为0.8mM。
[表1]
表1各种儿茶素类对F.n的硫化氢生成酶(半胱氨酸脱硫基酶)的抑制率
[表2]
表2各种儿茶素类对F.n的硫化氢生成酶(半胱氨酸脱硫基酶)的抑制率
[表3]
表3各种儿茶素类对F.n的甲硫氨酸酶的抑制率
2-3-2采用比色的EGCg对F.n、P.g和T.d的酶抑制试验
1-2-2的硫化氢生成酶抑制试验结果和1-2-3的甲硫氨酸酶抑制试验中,EGCg和氯化锌的抑制活性的比较结果示于图1。IC50示于表4。图1中,白色圆圈表示EGCg的结果,黑色圆圈表示氯化锌的结果。已知氯化锌具有口臭预防活性,是可期待硫化氢生成酶和甲硫氨酸酶的抑制的物质。由图1的结果可知,关于甲硫氨酸酶的抑制,无论是来源于F.n、P.g和T.d中的哪一种,氯化锌的抑制活性都高于EGCg的抑制活性。另一方面,关于硫化氢的生成,无论是来源于F.n、P.g和T.d中的哪一种,EGCg的抑制活性都高于氯化锌的抑制活性。由表4也能看出同样的结论。
[表4]
表4来源于具核梭杆菌JCM8532、牙龈卟啉单胞菌W83、齿垢密螺旋体ATCC35405的粗酶液的IC50(比色分析)
2-3-3采用气相色谱的EGCg对F.n、P.g和T.d的酶抑制试验
1-2-4的硫化氢生成酶抑制试验和1-2-5的甲硫氨酸酶抑制试验结果示于图2。IC50示于表5。由图2的结果可知,关于甲硫氨酸酶的抑制,无论是来源于F.n、P.g和T.d中的哪一种,氯化锌的抑制活性都高于EGCg的抑制活性。另一方面,关于硫化氢的生成,无论是来源于F.n、P.g和T.d中的哪一种,EGCg的抑制活性都高于氯化锌的抑制活性。由表5也能看出同样的结论。
[表5]
表5来源于具核梭杆菌JCM8532、牙龈卟啉单胞菌W83、齿垢密螺旋体ATCC35405的粗酶液的IC50(气相色谱分析)
2-4实施例1的总结
由以上结果可知:儿茶素、特别是其中的GCg、Cg、ECg和EGCg具有硫化氢生成酶抑制活性。特别是对于作为硫化氢生成酶的一种的半胱氨酸脱硫基酶具有显著的抑制活性。而且EGCg的活性远高于氯化锌的活性。EGCg的硫化氢生成酶抑制活性在来源于P.g和T.d的粗酶液中特别高,另外,在来源于F.n的粗酶液中可以观察到活性。
实施例2
通过常规方法制造含有本发明的由儿茶素类制成的硫化氢生成酶抑制剂的牙膏、口臭用喷雾、含片、口香糖、糖果、软糖、饮料。以下所示为它们的配方。本发明的产品不受到这些配方的限制。
牙膏的配方
口臭用喷雾的配方
含片的配方
口香糖的配方
糖果的配方
软糖的配方
饮料的配方
本申请主张基于2009年9月3日提出申请的日本专利申请号2009-203689的优先权,引用其内容作为本申请的一部分。
Claims (5)
1.一种硫化氢生成酶抑制剂,其特征在于,以儿茶素类作为有效成分。
2.如权利要求1所述的硫化氢生成酶抑制剂,其特征在于,儿茶素类是(-)-表儿茶素没食子酸酯、(-)-表没食子儿茶素没食子酸酯、(-)-没食子儿茶素没食子酸酯、(-)-儿茶素没食子酸酯的任一种或它们的混合物。
3.一种硫化氢生成酶抑制剂,其特征在于,以(-)-表儿茶素没食子酸酯、(-)-表没食子儿茶素没食子酸酯、(-)-没食子儿茶素没食子酸酯、(-)-儿茶素没食子酸酯的任一种或它们的混合物作为有效成分,抑制来源于具核梭杆菌(Fusobacterium nucleatum)、牙龈卟啉单胞菌(Porphyromonasgingivalis)或齿垢密螺旋体(Treponema denticola)的硫化氢生成酶。
4.一种饮食品,其特征在于,含有权利要求1~3中的任一项所述的硫化氢生成酶抑制剂。
5.一种口腔用组合物,其特征在于,含有权利要求1~3中的任一项所述的硫化氢生成酶抑制剂。
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CN108567773A (zh) * | 2018-04-23 | 2018-09-25 | 山东大学 | 一种用于清除口臭的口腔喷剂及其制备方法 |
CN108635378A (zh) * | 2018-04-23 | 2018-10-12 | 山东大学 | 一种用于日常口腔健康维护的口腔喷剂及其制备方法 |
CN109043383A (zh) * | 2018-09-05 | 2018-12-21 | 旌德县板桥现代渔业家庭农场 | 一种醋化乌龟蛋及其制备方法 |
CN116041100A (zh) * | 2023-02-24 | 2023-05-02 | 东北农业大学 | 一种降低堆肥中硫化氢释放量的方法 |
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CN108567773A (zh) * | 2018-04-23 | 2018-09-25 | 山东大学 | 一种用于清除口臭的口腔喷剂及其制备方法 |
CN108635378A (zh) * | 2018-04-23 | 2018-10-12 | 山东大学 | 一种用于日常口腔健康维护的口腔喷剂及其制备方法 |
CN108567773B (zh) * | 2018-04-23 | 2020-04-17 | 山东大学 | 一种用于清除口臭的口腔喷剂及其制备方法 |
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