UJI AKTIVITAS EKSTRAK KOLAGEN KULIT IKAN SALMON (Salmo salar) SEBAGAI INHIBITOR ACE1 BERDASARKAN STUDI IN VITRO DAN IN SILICO - Selmi Fiqhi Khoiriah, - (2022)
Penggunaan inhibitor ACE1 dalam pengobatan COVID-19 pada pasien berkomorbid telah diteliti menunjukkan efek samping. Kulit ikan salmon sebagai produk samping industri perikanan memiliki kandungan protein kolagen cukup tinggi dan berpotensi menjadi alternatif penemuan inhibitor ACE1 baru berbasis peptida. Penelitian ini bertujuan untuk menguji aktivitas kolagen kulit ikan salmon (Salmo salar) sebagai inhibitor ACE1 berdasarkan studi in vitro dan in silico. Metode penelitian in vitro meliputi ekstraksi kolagen larut asam, karakterisasi menggunakan FTIR, UV, XRD, dan SDS-PAGE, serta penentuan inhibisi terhadap ACE1. Adapun metode pada studi in silico yang digunakan yaitu molecular docking. Ekstraksi kolagen memiliki kandungan protein 95,32±2,2 %. Spektrum FTIR mendeteksi gugus fungsi amida (A, B, I, II, III) dan imina prolin/hidroksiprolin yang mengindikasikan serapan khas kolagen. Karakterisasi UV menunjukkan serapan khas kolagen pada 230 nm. Berdasarkan data XRD dan SDS-PAGE, jenis rantai pada sampel adalah α-heliks dan rantai α. Pengujian inhibisi ACE1 dari sampel didapat sebesar 11,42%; 67,94%; 74,79%; 83,67%; dan 84,94% berturut-turut pada konsentrasi 10, 250, 500, 1000, dan 2000 ppm. Adapun studi in silico menunjukkan adanya interaksi antara ACE1 dan kolagen di luar sisi aktif dan afinitas sebesar -213,89 kkal/mol. Sementara itu, peptida aktif kolagen menunjukkan interaksi pada sisi aktif ACE1 dengan afinitas lebih baik dari lisinopril pada tiga peptida, yaitu WF (Trp-Phe), WYT (Trp-TyrThr), dan PDPF (Pro-Asp-Pro-Phe) berturut-turut 9,1; 9,1; dan 8,8 kkal/mol. Dapat disimpulkan bahwa kolagen kulit ikan salmon memiliki aktivitas inhibisi ACE1 dengan prediksi mekanisme inhibisi non-kompetitif. Sebaliknya, peptida aktif dari kolagen kulit ikan salmon diprediksi memiliki potensi sebagai inhibitor kompetitif terhadap ACE1. Kata Kunci: ACE1, in vitro, in silico, Kulit Ikan Salmon, Kolagen ABSTRACT The use of ACE1 inhibitors in COVID-19's comorbid patient treatment has been studied to have adverse effects. Salmon skin, as a by-product of the fishing industry, has potential due to its high collagen protein content and can be an alternative source for discovering new peptide-based ACE1 inhibitors. This study aims to determine the collagen activity of salmon skin (Salmo salar) as an ACE1 inhibitor based in vitro and in silico. In vitro research methods include extraction of acid-soluble collagen, characterization using FTIR, UV, XRD, and SDS-PAGE, and determination of ACE1 inhibition. The method used in the in silico study is molecular docking. The collagen extraction process gives protein content of 95.32±2.2 (%). FTIR analysis showed the spectrum of amide functional groups (A, B, I, II, III) and imine proline/hydroxyproline typical of collagen. UV characterization showed a typical absorption of collagen at 230 nm. Based on XRD and SDS-PAGE data, the sample chain type is alpha helix and alpha chain. The ACE1 inhibition test of the sample was 11.42%; 67.94%; 74.79%; 83.67%; and 84.94% at concentrations of 10, 250, 500, 1000, and 2000 ppm, respectively. The in silico study showed interactions between ACE1 and collagen outside the active site and an affinity of -213.89 kcal/mol. Meanwhile, the active peptide of collagen showed interactions at the active site of ACE1 with better affinity than lisinopril for three peptides, namely WF (Trp-Phe), WYT (Trp-Tyr- Thr ), and PDPF (ProAsp-Pro-Phe) 9.1; 9.1; and 8.8 kcal/mol. It can be concluded that salmon skin collagen has ACE1 inhibitory activity with a mechanism that is predictable as non-competitive inhibition. In contrast, active peptides from salmon skin collagen were predicted to have potential as competitive inhibitors of ACE1. Keywords: ACE1, collagen, in vitro, in silico, salmon skin