Screening of Angiotensin-I Converting Enzyme Inhibitory Peptide Derived from Taiwan Red Quinoa (Chenopodium formosanum) using Tandem Bioassay-Guided Fractionation

Suleman, Dininurilmi Putri and Prof. Jue-Liang Hsu,, Ph.D and Prof. Dr. Ir. Harijono,, M.App.Sc (2020) Screening of Angiotensin-I Converting Enzyme Inhibitory Peptide Derived from Taiwan Red Quinoa (Chenopodium formosanum) using Tandem Bioassay-Guided Fractionation. Magister thesis, Universitas Brawijaya.

Abstract

Angiotensin I-converting enzyme (ACE) adalah peptidil dipeptidase yang mengkatalisis konversi dekapeptida angiotensin I menjadi oktapeptide angiotensin II, agen vasokonstriktor, dengan menghilangkan terminal karboksi dipeptida. ACE telah lama dikenal sebagai bagian penting dari renin- sistem angiotensin yang mengatur tekanan darah, dan inhibitor ACE adalah penting untuk pengobatan hipertensi. Quinoa Merah Taiwan (Chenopodium formosanum) merupakan salah satu tanaman serealia tradisional Taiwan. Dia mengandung sumber protein yang berpotensi memiliki bioaktivitas untuk hipertensi perlakuan. Protein yang terkandung lebih tinggi dari tanaman sereal umum lainnya seperti seperti gandum, oat atau nasi. Penelitian ini bertujuan untuk mengeksplorasi aktivitas biologis untuk menghambat ACE dari hidrolisat protein Red Quinoa. Protein yang diekstraksi dari Red Quinoa dihidrolisis oleh berbagai protease, yaitu termolisin, pepsin, tripsin, dan -kimotripsin. Itu Aktivitas penghambatan ACE dari hidrolisat yang dihasilkan dievaluasi menggunakan: uji penghambatan ACE in vitro. Hasilnya menunjukkan bahwa thermolysin hidrolisat menunjukkan aktivitas penghambatan tertinggi, sebesar 77,67%. Fraksinasi yang dipandu bioassay tandem menggunakan Pertukaran Kation Kuat (SCX) dan Kromatografi Cair Kinerja Tinggi Fase Terbalik (RP- HPLC) dilakukan untuk mengetahui pecahan terbaik dengan ACE . tertinggi aktivitas penghambatan. F4 (87,06%) dari SCX dan F4.3 (89,30%) dari Tandem Fraksinasi SCX-RPHPLC menunjukkan aktivitas penghambatan terbaik, masing-masing. F4.3 selanjutnya diurutkan menggunakan kromatografi cair spektrometri massa tandem digabungkan dengan de novo sequencing untuk mendapatkan beberapa peptida potensial. Peptida Val-Tyr-Leu-Ala-Glu-Leu-His-Phe (VF-8), Leu- Gly-Ala-Val-Pro-Pro-Arg-Tyr (LY-8) dan Ile-Ala-Arg-Asp-Ser-Ala-Ala- Val-Phe (IF-9) diidentifikasi dengan skor tinggi dan nilai IC50 adalah ditentukan sebagai 24,26 ± 1,63 M, 30,48 ± 1,39 M, dan 72,57 ± 3,5 M, masing-masing. Studi kinetik dan simulasi molekuler docking mengungkapkan bahwa VF-8 adalah inhibitor kompetitif. Oleh karena itu, kita dapat menyimpulkan AKU AKU AKU bahwa penelitian ini adalah laporan pertama tentang peptida penghambat ACE yang berasal dari Taiwan Red Quinoa dan mungkin bermanfaat untuk mencegah hipertensi dan pengembangan pangan fungsional

English Abstract

Angiotensin I-converting enzyme (ACE) is a peptidyl dipeptidase that catalyzes the conversion of the decapeptide angiotensin I to the octapeptide angiotensin II, the vasoconstrictor agent, by removing a carboxy-terminal dipeptide. ACE has long been known to be a crucial part of the renin- angiotensin system that regulates blood pressure, and ACE inhibitors are important for the treatment of hypertension. Taiwan Red Quinoa (Chenopodium formosanum) is one of the Taiwan traditional cereal plant. It contains a protein source that potentially has bioactivity for hypertension treatment. Its protein contained is higher than other general cereal plant such as wheat, oat or rice. This study aims to explore a biological activity to inhibit ACE from Red Quinoa protein hydrolysate. Proteins extracted from Red Quinoa were hydrolyzed by various proteases, namely thermolysin, pepsin, trypsin, and α-chymotrypsin. The ACE inhibitory activities of the resulting hydrolysates were evaluated using in vitro ACE inhibitory assay. The result indicated that thermolysin hydrolysate showed the highest inhibitory activity, as high as 77.67%. Tandem bioassay-guided fractionations using Strong Cation Exchange (SCX) and Reverse Phase-High Performance Liquid Chromatography (RP- HPLC) were performed to find out the best fractions with the highest ACE inhibitory activity. F4 (87.06%) from SCX and F4.3 (89.30%) from Tandem SCX-RPHPLC fractionations showed the best inhibition activity, respectively. F4.3 was further sequenced using liquid chromatography tandem mass spectrometry coupled with de novo sequencing to get some potential peptides. Peptide Val-Tyr-Leu-Ala-Glu-Leu-His-Phe (VF-8), Leu- Gly-Ala-Val-Pro-Pro-Arg-Tyr (LY-8) and Ile-Ala-Arg-Asp-Ser-Ala-Ala- Val-Phe (IF-9) were identified with high score and the IC50 values were determined as 24.26 ± 1.63 μM, 30.48 ± 1.39 μM, and 72.57 ± 3.5 μM, respectively. The kinetic study and the simulation molecular docking revealed that VF-8 is a competitive inhibitor. Therefore, we can conclude that this study is the first report about ACE inhibitory peptides derived from Taiwan Red Quinoa and it may be beneficial for preventing hypertension and functional food development

Item Type:	Thesis (Magister)
Identification Number:	0420090001
Uncontrolled Keywords:	Angiotensin I-converting enzyme, active peptides, Taiwan Red Quinoa, protein hydrolysate.
Subjects:	500 Natural sciences and mathematics > 570 Biology
Divisions:	S2/S3 > Magister Biologi, Fakultas MIPA
Depositing User:	soegeng sugeng
Date Deposited:	20 Jul 2022 07:15
Last Modified:	01 Oct 2024 02:30
URI:	http://repository.ub.ac.id/id/eprint/192424

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