Pratiwi Arby'in, Hakim Thoyib R, Abidin Mohammad Z, Fitriyanto Nanung A, Jamhari Jamhari, Rusman Rusman, Erwanto Yuny
Department of Animal Products Technology, Faculty of Animal Sciences, Universitas Gadjah Mada, Jl. Fauna No. 3, Bulaksumur, Yogyakarta 55281, Indonesia.
Division of Halal Materials Development, Institute for Halal Industry and System, Universitas Gadjah Mada, Jl. Kaliurang km. 4.5, Bulaksumur, Yogyakarta 55281, Indonesia.
Vet World. 2021 Jan;14(1):161-167. doi: 10.14202/vetworld.2021.161-167. Epub 2021 Jan 21.
Angiotensin-converting enzyme (ACE) is one of the inhibitory enzymes isolated from animals for the treatment of hypertension. ACE inhibitor (ACE-I) peptides can be obtained by hydrolyzing proteins from various animal tissues, including muscle and connective tissues. However, the study on ACE-I activity from collagen of goat skin has not been conducted. This study explores the potency of collagen from goat skin as a source of an antihypertensive agent through ACE inhibition. Thermolysin will hydrolyze collagen and produce the peptide classified antihypertensive bioactive peptides. This study aimed to determine the potential of thermolysin to hydrolyze collagen of goat skin for ACE-I peptide production and to identify the production of ACE-I peptides.
Collagen from goat skin was hydrolyzed with thermolysin and incubated at 37°C for 1 h. Molecular weight (MW) evaluation was performed by SDS PAGE; fractionation peptides at <5 kDa, 3-5 kDa, and <3 kDa were performed by ultrafiltration and ACE-I activity determined by IC measurement.
Collagen was hydrolyzed by thermolysin, resulting in protein with MW of 117.50-14.60 kDa. The protein content of fractionation at >5 kDa was 3.93±0.72 mg/mL, content of 3-5 kDa was 3.81±0.68 mg/mL, and that of <3 kDa was 2.33±0.38 mg/mL. Fractionation was performed 3 times and one of the results was selected for the ACE-I test. The selected fraction was tested by IC measurement with three repetitions and it showed an average enzyme activity at 0.83 mg/mL or 82.94 mg/mL.
Thermolysin hydrolysis of collagen from goat skin showed the potential to produce bioactive peptides, such as ACE-I.
血管紧张素转换酶（ACE）是从动物体内分离出的用于治疗高血压的抑制酶之一。ACE抑制剂（ACE-I）肽可通过水解包括肌肉和结缔组织在内的各种动物组织中的蛋白质获得。然而，尚未对山羊皮胶原蛋白的ACE-I活性进行研究。本研究通过ACE抑制作用探索山羊皮胶原蛋白作为抗高血压药物来源的潜力。嗜热菌蛋白酶将水解胶原蛋白并产生分类为抗高血压生物活性肽的肽。本研究旨在确定嗜热菌蛋白酶水解山羊皮胶原蛋白以生产ACE-I肽的潜力，并鉴定ACE-I肽的产生。
用嗜热菌蛋白酶水解山羊皮胶原蛋白，并在37°C孵育1小时。通过SDS-PAGE进行分子量（MW）评估；通过超滤对分子量小于5 kDa、3-5 kDa和小于3 kDa的肽进行分级分离，并通过IC测量确定ACE-I活性。
胶原蛋白被嗜热菌蛋白酶水解，产生分子量为117.50-14.60 kDa的蛋白质。大于5 kDa分级分离的蛋白质含量为3.93±0.72 mg/mL，3-5 kDa的含量为3.81±0.68 mg/mL，小于3 kDa的含量为2.33±0.38 mg/mL。分级分离进行3次，选择其中一个结果进行ACE-I测试。所选级分通过IC测量进行三次重复测试，其平均酶活性为0.83 mg/mL或82.94 mg/mL。
嗜热菌蛋白酶水解山羊皮胶原蛋白显示出产生生物活性肽（如ACE-I）的潜力。
