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Biochemical Studies on Halocynine, a New Betaine Isolated from the Muscle of an Ascidian Halocynthia roretzi.

对从海鞘类 Halocynthia roretzi 肌肉中分离出的新型甜菜碱 Halocynine 进行生化研究。

基本信息

批准号：
61560229
负责人：
WATANABE Katsuko
金额：
$ 1.22万
依托单位：
Faculty of Agriculture, University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1986
资助国家：
日本
起止时间：
1986 至 1987
项目状态：
已结题

项目摘要

The present studies were conducted to develop an HPLC method for determination of halocynine, a new betaine isolated from the muscle of an ascidian Halocynthia roretzi, and to survey its distribution in the tissues of the ascidian and in the related species. Biosynthesis of halocynine in H. roretzi was also investigated with tracer technics. Results obtained were summarized as follows.1. The authentic halocynine was esterified with UV-absorbing ,p-dibromoacetophenone and subjected to an HPLC using Shim-pack FLC-CN column and 5 mM KH_2PO_4 buffer containing 5 % CH_3CN (pH 2.0) with an ultraviolet detector operater at 262 nm. Halocynine p-bromophenacyl ester was well separated from the ester of glycinbetaine and other UV-absorbing substances which were abundant in the ascidian muscle. The linear relationship was obtained between the peak areas of halocynine ester and the amounts of halocynine up to 300 nmol.2. More than 50 mg of halocynine per 100 g was detected in the muscle of H. roretzi throughout the year, but its content was high in winter and low in summer. Small amounts of halocynine were found in the liver and gills but not in the genital gland and tunica. Halocynine ranging from a trace amount to 30 mg per 100 g was also detected in the muscle, liver, gonad and gills of Styela plicata and in the whole bodies of Botrylloides violaceus, Pseudoistoma antinboja and Ciona intestinalis.3. After ^<14>C-lysine was injected into the muscle of H. roretzi, the animals were kept in a seawater tank for 10 days. Then, a picric acid extract was prepared from the muscle and subjected to an ion exchange column chromatography to isolate halocynine, which showed a radioactivity. The radioactivity migrated with the spots of isolated halocynine on TLC with four different solvent systems. These results clearly show that halocynine is biosynthesized from lysine in H. roretzi.

建立了从海鞘肌中分离得到的一种新型甜菜碱盐碱碱的HPLC测定方法，并对其在海鞘及近缘种组织中的分布进行了研究。用示踪技术研究了卤代碱的生物合成。所得结果总结如下：1。用吸紫外的对二溴苯乙酮对真卤代盐进行酯化，采用Shim-pack FLC-CN柱和含5% CH_3CN （pH 2.0）的5 mM KH_2PO_4缓冲液进行高效液相色谱（HPLC），紫外检测器为262 nm。卤素对溴苯酰酯与甘糖碱酯及其他大量存在于海鞘肌中的紫外线吸收物质有较好的分离。在300 nmol / 2以内，卤代碱酯峰面积与卤代碱用量呈线性关系。海参肌肉中盐碱碱含量全年均在50 mg / 100g以上，但冬季含量高，夏季含量低。在肝脏和鳃中发现了少量的盐碱碱，但在生殖腺和膜中没有发现。在海参的肌肉、肝脏、性腺和鳃中，以及在堇菜、抗花蓟马和花蓟马的全身中，也检测到每100克盐碱含量从微量到30毫克不等。将c -赖氨酸注射到罗氏鼠肌肉中，在海水池中饲养10天。然后，从肌肉中提取苦味酸提取物，并进行离子交换柱层析，分离出具有放射性的卤代盐碱。在4种不同的溶剂体系下，放射性随分离的卤代碱在TLC上的斑点而迁移。这些结果清楚地表明，盐氨酸是由水蛭中的赖氨酸生物合成的。