Laboratoire de Genie Enzymatique et de Microbiologie-Ecole Nationale d'Ingenieurs de Sfax, Sfax, Tunisia.
J Agric Food Chem. 2010 Mar 24;58(6):3840-6. doi: 10.1021/jf904300q.
The angiotensin I-converting enzyme (ACE) inhibitory activities of protein hydrolysates prepared from cuttlefish (Sepia officinalis) proteins by treatment with various bacterial proteases were investigated. The hydrolysate generated by the crude enzyme from Bacillus mojavensis A21 displayed the highest ACE inhibitory activity, and the higher inhibition activity (87.11 +/- 0.92% at 2 mg/mL) was obtained with hydrolysis degree of 16%. This hydrolysate was fractionated by size exclusion chromatography on a Sephadex G-25 into eight major fractions (P(1)-P(8)). Fraction P(6), which exhibited the highest ACE inhibitory activity, was then fractionated by reversed-phase high performance liquid chromatography (RP-HPLC). Eleven ACE inhibitory peptides were isolated, and their molecular masses and amino acids sequences were determined using ESI-MS and ESI-MS/MS, respectively. The structures of the most potent peptides were identified as Ala-His-Ser-Tyr, Gly-Asp-Ala-Pro, Ala-Gly-Ser-Pro and Asp-Phe-Gly. The first peptide displayed the highest ACE inhibitory activity with an IC(50) of 11.6 microM. The results of this study suggest that cuttlefish protein hydrolysates are a good source of ACE inhibitory peptides.
采用不同细菌蛋白酶水解乌贼蛋白制备蛋白水解物，研究其对血管紧张素转化酶（ACE）的抑制活性。由莫拉氏芽孢杆菌 A21 粗酶制备的水解物对 ACE 的抑制活性最高，在 2mg/mL 时水解度为 16%时具有更高的抑制活性（87.11±0.92%）。该水解物通过 Sephadex G-25 分子筛层析进一步分离成 8 个主要馏分（P(1)-P(8)）。具有最高 ACE 抑制活性的 P(6)馏分，然后通过反相高效液相色谱（RP-HPLC）进一步分离。分离得到 11 个 ACE 抑制肽，分别采用 ESI-MS 和 ESI-MS/MS 确定其分子量和氨基酸序列。最有效的肽的结构被鉴定为 Ala-His-Ser-Tyr、Gly-Asp-Ala-Pro、Ala-Gly-Ser-Pro 和 Asp-Phe-Gly。第一个肽对 ACE 的抑制活性最高，IC(50)为 11.6μM。本研究结果表明，乌贼蛋白水解物是 ACE 抑制肽的良好来源。
