Krichen Fatma, Sila Assaâd, Caron Juliette, Kobbi Sabrine, Nedjar Naima, Miled Nabil, Blecker Christophe, Besbes Souhail, Bougatef Ali
Laboratoire d'Amélioration des Plantes et Valorisation des Agroressources Université de Sfax Sfax Tunisia.
Institut Régional de Recherche en Agroalimentaire et Biotechnologie: Charles Violette Equipe ProBioGEM, Université de Lille 1 France.
Eng Life Sci. 2018 Jul 15;18(9):682-691. doi: 10.1002/elsc.201800045. eCollection 2018 Sep.
The effect of enzymatic hydrolysis by Savinase on the interfacial properties and antihypertensive activity of shrimp waste proteins was evaluated. The physicochemical characterization, interfacial tension, and surface characteristics of shrimp waste protein hydrolysates (SWPH) using different enzyme/substrate (E/S) (SWPH (SWPH using E/S = 5), SWPH (SWPH using E/S = 15), and SWPH (SWPH using E/S = 40)) were also studied. SWPH, SWPH, and SWPH had an isoelectric pH around 2.07, 2.17, and 2.54 respectively. SWPH exhibited the lowest interfacial tension (68.96 mN/m) followed by SWPH (69.36 mN/m) and SWPH (70.29 mN/m). The in vitro ACE inhibitory activity of shrimp waste protein hydrolysates showed that the most active hydrolysate was obtained using an enzyme/substrate of 15 U/mg (SWPH). SWPH had a lower IC value (2.17 mg/mL) than that of SWPH and SWPH (3.65 and 5.7 mg/mL, respectively). This hydrolysate was then purified and characterized. Fraction F1 separated by Sephadex G25 column which presents the best ACE inhibition activity was then separated by reversed-phase high performance liquid chromatography. Four ACE inhibitory peptides were identified and their molecular masses and amino acid sequences were determined using ESI-MS and ESI-MS/MS, respectively. The structures of the most potent peptides were SSSKAKKMP, HGEGGRSTHE, WLGHGGRPDHE, and WRMDIDGDIMISEQEAHQR. The structural modeling of anti-ACE peptides from shrimp waste through docking simulations results showed that these peptides bound to ACE with high affinity.
评估了萨维诺酶（Savinase）酶解对虾壳废弃物蛋白界面性质和降压活性的影响。还研究了不同酶/底物比（E/S）的虾壳废弃物蛋白水解物（SWPH）（SWPH（E/S = 5时的SWPH）、SWPH（E/S = 15时的SWPH）和SWPH（E/S = 40时的SWPH））的物理化学特性、界面张力和表面特性。SWPH、SWPH和SWPH的等电点pH分别约为2.07、2.17和2.54。SWPH表现出最低的界面张力（68.96 mN/m），其次是SWPH（69.36 mN/m）和SWPH（70.29 mN/m）。虾壳废弃物蛋白水解物的体外ACE抑制活性表明，使用15 U/mg的酶/底物（SWPH）获得了活性最高的水解物。SWPH的IC值（2.17 mg/mL）低于SWPH和SWPH（分别为3.65和5.7 mg/mL）。然后对该水解物进行纯化和表征。通过Sephadex G25柱分离出具有最佳ACE抑制活性的F1级分，然后通过反相高效液相色谱进一步分离。鉴定出四种ACE抑制肽，并分别使用电喷雾电离质谱（ESI-MS）和电喷雾电离串联质谱（ESI-MS/MS）测定了它们的分子量和氨基酸序列。最有效的肽的结构为SSSKAKKMP、HGEGGRSTHE、WLGHGGRPDHE和WRMDIDGDIMISEQEAHQR。通过对接模拟对虾壳废弃物中的抗ACE肽进行结构建模的结果表明，这些肽与ACE具有高亲和力。
