Graduate School of Advanced Technology and Science, Tokushima University, 2-1 Minamijosanjima-cho, Tokushima 770-8506, Japan.
Graduate School of Technology, Industrial and Social Science, Tokushima University, 2-1 Minamijosanjima-cho, Tokushima 770-8513, Japan.
Nutrients. 2022 Mar 29;14(7):1422. doi: 10.3390/nu14071422.
Cockscomb hydrolysate was found to have neurite outgrowth-promoting activity in PC12 cells. To investigate the neurite outgrowth-promoting compounds derived from cockscomb hydrolysate, bioassay-guided purification was carried out. Purified active fractions were obtained by liquid-liquid partition, followed by column chromatography. High-performance liquid chromatography and proton nuclear magnetic resonance analyses of the purified active fractions clarified that the main compounds are threonine, alanine, valine, and methionine. By screening for 20 kinds of amino acids, it was shown that valine and methionine, but not threonine and alanine, have neurite outgrowth-promoting activity. The results of activity evaluation of the mixture of amino acids indicated that alanine enhanced the activity of valine and that the mixture of valine and methionine showed a higher ratio of neurite formation than did each of them alone. On the other hand, dipeptides formed by valine and methionine showed weak neurite outgrowth-promoting activity. A mixture of threonine, alanine, valine, and methionine at the same concentrations as those in cockscomb hydrolysate showed neurite outgrowth-promoting activity comparable to that of cockscomb hydrolysate although threonine, alanine, valine, and methionine alone did not show activity at their concentrations in cockscomb hydrolysate. Therefore, the strong neurite outgrowth-promoting activity of cockscomb hydrolysate was considered to be due to the synergistic effect of threonine, alanine, valine, and methionine.
鸡冠水解物被发现具有促进 PC12 细胞突起生长的活性。为了研究源自鸡冠水解物的促进突起生长的化合物，进行了基于生物测定的纯化。通过液液分配和柱层析获得了纯化的活性级分。通过高效液相色谱和质子核磁共振分析对纯化的活性级分进行了阐明，主要化合物为苏氨酸、丙氨酸、缬氨酸和蛋氨酸。通过对 20 种氨基酸进行筛选，表明缬氨酸和蛋氨酸具有促进突起生长的活性，而不是苏氨酸和丙氨酸。氨基酸混合物的活性评价结果表明，丙氨酸增强了缬氨酸的活性，并且缬氨酸和蛋氨酸的混合物比它们各自单独显示出更高的突起形成比例。另一方面，由缬氨酸和蛋氨酸形成的二肽显示出较弱的促进突起生长的活性。在与鸡冠水解物中相同浓度的苏氨酸、丙氨酸、缬氨酸和蛋氨酸的混合物表现出与鸡冠水解物相当的促进突起生长的活性，尽管苏氨酸、丙氨酸、缬氨酸和蛋氨酸本身在鸡冠水解物中的浓度下没有活性。因此，鸡冠水解物的强烈促进突起生长的活性被认为是由于苏氨酸、丙氨酸、缬氨酸和蛋氨酸的协同作用。
