Simultaneous analysis of free amino acids and taurine-related compounds in deep-sea mussel tissues using reversed-phase HPLC
Simultaneous analysis of free amino acids and taurine-related compounds in deep-sea mussel tissues using reversed-phase HPLC
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DOI：
10.1007/s12562-017-1143-8
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发表时间：
2018-01-01
影响因子：
1.9
通讯作者：
Inoue, Koji
中科院分区：
文献类型：
--
作者：
Nagasaki, Toshihiro;Koito, Tomoko;Inoue, Koji
牛磺酸和亚牛磺酸大量存在于软体动物组织中；然而，它们的生物合成途径尚未得到充分研究。在本研究中，我们建立了一种同时检测牛磺酸相关化合物的方法，包括牛磺酸、亚牛磺酸、半胱氨酸、半胱氨酸亚磺酸（CSA）、胱硫醚、γ-氨基丁酸和β-丙氨酸以及17种游离氨基酸，在含氨溶液中使用苯硫基氨基甲酰衍生化，然后注射到反相高效液相色谱中。使用这种方法，我们分析了热液喷口特定贻贝Bathymodiolus septemdierum的主要组织（鳃、外套膜、内收肌、足和消化腺）。鳃和消化腺被认为是牛磺酸/亚牛磺酸生物合成的主要器官，因为牛磺酸和亚牛磺酸水平往往较高，并且在这两个组织中检测到它们的前体，即半胱氨酸、半胱氨酸和CSA（仅在消化腺中）。在消化腺中检测到的胱硫醚水平相对较高，而在其他组织中检测到的水平较低，表明其作为牛磺酸/亚牛磺酸生物合成的储存材料的作用。研究发现内收肌富含游离氨基酸，这主要是由于富含非硫氨基酸。
Taurine and hypotaurine are abundantly present in molluskan tissues; however, the pathways for their biosynthesis have not yet been well investigated. In this study, we established a method for simultaneous detection of taurine-related compounds, including taurine, hypotaurine, cysteic acid, cysteine sulfinic acid (CSA), cystathionine, gamma-aminobutyric acid, and beta-alanine along with 17 free amino acids, using phenylthiocarbamyl derivatization in ammonia-containing solution, followed by injection into reversed-phase high-performance liquid chromatography. Using this method, we analyzed the major tissues (gill, mantle, adductor muscle, foot, and digestive gland) of the hydrothermal-vent-specific mussel Bathymodiolus septemdierum. The gill and digestive gland were suggested to be the major organs for taurine/hypotaurine biosynthesis because taurine and hypotaurine levels tended to be high, and their precursors, i.e., cysteine, cysteic acid, and CSA (only in digestive gland), were detected in these two tissues. Cystathionine was detected at relatively high level in digestive gland and in other tissues at low levels, suggesting its role as storage material for taurine/hypotaurine biosynthesis. Adductor muscle was found to be rich in free amino acids, mainly due to abundance of non-sulfur-containing amino acids.