Yokoyama K, Chiba H, Yoshikawa M
Nippon Synthetic Chemical Industry Co., Ltd., Osaka, Japan.
Biosci Biotechnol Biochem. 1992 Oct;56(10):1541-5. doi: 10.1271/bbb.56.1541.
Dried bonito (Katsuobusi), a Japanese traditional seasoning made of bonito muscle was hydrolyzed by various proteases and the inhibitory activity of the hydrolyzates for angiotensin I-converting enzyme (ACE) [EC 3.4.15.1] was measured. Among the digests, thermolysin digest showed the most potent inhibitory activity. Eight inhibitory peptides were isolated from the digest using HPLC. The amino acid sequences of inhibitory peptides were Ile-Lys-Pro-Leu-Asn-Tyr, Ile-Val-Gly-Arg-Pro-Arg-His-Gln-Gly, Ile-Trp-His-His-Thr, Ala-Leu-Pro-His-Ala, Phe-Gln-Pro, Leu-Lys-Pro-Asn-Met, Ile-Tyr, and Asp-Tyr-Gly-Leu-Tyr-Pro. By searching for the sequence homology in many proteins, four of them were found in the primary structure of actin. Asp-Met-Ile-Pro-Ala-Gln-Lys was obtained from the boiling water extract of dried bonito and this peptide was found in the primary structure of creatine kinase. Fragments of these peptides were prepared by further enzymatic digestion or chemical synthesis and their ACE-inhibitory activities were measured. Among them, Ile-Lys-Pro, Ile-Trp, Leu-Lys-Pro, and Leu-Tyr-Pro had higher inhibitory activity than their parental peptides. Ile-Lys-Pro suppressed the hypertensive activity of angiotensin I.
干鲣鱼（柴鱼）是一种由鲣鱼肌肉制成的日本传统调味料，用各种蛋白酶对其进行水解，并测定水解产物对血管紧张素I转换酶（ACE）[EC 3.4.15.1]的抑制活性。在这些消化产物中，嗜热菌蛋白酶消化产物显示出最强的抑制活性。使用高效液相色谱法从消化产物中分离出8种抑制肽。抑制肽的氨基酸序列为Ile-Lys-Pro-Leu-Asn-Tyr、Ile-Val-Gly-Arg-Pro-Arg-His-Gln-Gly、Ile-Trp-His-His-Thr、Ala-Leu-Pro-His-Ala、Phe-Gln-Pro、Leu-Lys-Pro-Asn-Met、Ile-Tyr和Asp-Tyr-Gly-Leu-Tyr-Pro。通过在许多蛋白质中搜索序列同源性，发现其中4种存在于肌动蛋白的一级结构中。Asp-Met-Ile-Pro-Ala-Gln-Lys是从干鲣鱼的沸水提取物中获得的，该肽存在于肌酸激酶的一级结构中。通过进一步的酶消化或化学合成制备这些肽的片段，并测定它们的ACE抑制活性。其中，Ile-Lys-Pro、Ile-Trp、Leu-Lys-Pro和Leu-Tyr-Pro的抑制活性高于其亲本肽。Ile-Lys-Pro抑制了血管紧张素I的高血压活性。
