Investigation on the formation and the determination of γ-glutamyl-β-alanylhistidine and related isopeptide in the macromolecular fraction of beef soup stock
Investigation on the formation and the determination of γ-glutamyl-β-alanylhistidine and related isopeptide in the macromolecular fraction of beef soup stock
复制标题
牛肉汤大分子组分中γ-谷氨酰-β-丙氨酰组氨酸及相关异肽的形成及测定
DOI：
10.1021/jf001095r
复制
发表时间：
2000
影响因子：
6.1
通讯作者：
T. Harada
中科院分区：
文献类型：
--
作者：
M. Kuroda;R. Ohtake;E. Suzuki;T. Harada
为了确认γ-谷氨酰-β-丙氨酰组氨酸和相关肽的形成，加热模型溶液（含酰胺的氨基酸和肌肽），并研究产物。光谱分析表明，天冬酰胺与肌肽反应的主要产物为β-谷氨酰-β-丙氨酰组氨酸，谷氨酰胺与肌肽反应的主要产物为γ-谷氨酰-β-丙氨酰组氨酸。此外，为了证实上述肽在食品加热过程中的增加，建立了测定食品蛋白质中这些异肽的HPLC方法。通过蛋白水解消化释放异肽，并使用Toyopack IC-SP柱通过固相萃取进行分馏。用异硫氰酸苯酯（PITC）衍生含有异肽的馏分，并使用十八烷基硅胶柱通过HPLC进行分离和定量。定量结果表明，加热牛肉汤原液中大分子组分中γ-谷氨酰-β-丙氨酰组氨酸异肽的增加具有B。
To confirm the formation of γ-glutamyl-β-alanylhistidine and related peptide, a model solution (amide-containing amino acids and carnosine) has been heated, and the products are investigated. Spectroscopical analysis indicates that the major product from asparagine and carnosine is β-aspartyl-β-alanylhistidine, and that from glutamine and carnosine is γ-glutamyl-β-alanylhistidine. Furthermore, to confirm the increase of the above peptides during the heating process of food, an HPLC method for the determination of these isopeptides in food protein is constructed. The isopeptides are liberated by proteolytic digestion and fractionated by solid-phase extraction using Toyopack IC-SP cartridges. The fraction containing the isopeptides is derivatized with phenylisothiocyanate (PITC) and separated and quantified by HPLC using an octadecyl−silica column. As a result of quantification, an increase of the γ-glutamyl-β-alanylhistidine isopeptide in the macromolecular fraction of heated beef soup stock solution has b...