Study on Serotonine Metabolism in Fish

鱼类血清素代谢的研究

• 批准号: 04454096
• 负责人: NAGAYAMA Fumio
• 金额: $ 2.69万
• 依托单位: Tokyo University of Fisheries
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454096/
• 关键词: fish; tryptophan; serotonine; melatonine; enzyme; 酵素系

Distribution and properties of several enzymes which are involved in serotonine pathway in fish were investigated. The method of matabolte analysis was also examined.1. Triptophan hydroxylase : Higher activity of this enzyme was observed in fish liver, whileitwas reported that the activity is high in brain tissue in mammals. However, the activity varied depending on fish species. The molecular weight of the skipjack enzyme was 290,000(oligomer), the optimum pH was 8.0, and Km for tryptopha was 7.0 x 10^<-5>M.2. Aromatic amino acid decarboxylase : The moleclar weight of the skipjack enzyme was 220,000(dimer), the optimum pH was 7.0, and Km for hydroxytroptohan was 6.97 x 10^<-4>M.The substrate specificity was rather wide.3. Monoamine oxidase : There were 2 types of the enzyme in fish, and the patern of the activation and inhibition by inorganic ions differed each other.4. Method of metabolite analysis : HPLC system for analysis of metabolites in serotonine pathway was examined. The condition was as follows : (a) reverse phase, JASCO SIL C18 coolumn, (b) mobile phase, a mixture of 10mM sodium acetate soltion and acetonitrile, (c) flow rate, 1ml/min, (d) monitor, fluoressence, Ex 285 nm, Em 345 nm. By changing the ratio of actate solution and acetonitrile, we were able to determine the metabolites quantitatively (2 - 12 ng of each compound).

研究了鱼类中参与血清素途径的几种酶的分布和性质。并对代谢物分析方法进行了探讨。 1．色氨酸羟化酶：在鱼肝中观察到该酶的活性较高，而据报道，在哺乳动物的脑组织中该酶的活性较高。然而，活动因鱼类种类而异。鲣鱼酶的分子量为290,000(寡聚物)，最适pH为8.0，色氨酸Km为7.0×10^-5M.2。芳香族氨基酸脱羧酶：鲣鱼酶的分子量为220,000（二聚体），最适pH为7.0，对羟基海藻聚糖的Km为6.97×10^-4M，底物特异性较宽。3．单胺氧化酶：鱼类体内有2种酶，其被无机离子激活和抑制的模式各不相同。 4．代谢物分析方法：检查了用于分析血清素途径代谢物的HPLC系统。条件如下：(a)反相，JASCO SIL C18冷却，(b)流动相，10mM乙酸钠溶液和乙腈的混合物，(c)流速，1ml/min，(d)监测器，荧光，Ex 285 nm，Em 345 nm。通过改变乙酸盐溶液和乙腈的比例，我们能够定量测定代谢物（每种化合物 2 - 12 ng）。