Screening and Characterization of Angiotensin-I Converting Enzyme Inhibitory Peptides Derived from Enzymatic Hydrolysate of Bitter Melon Seed Proteins

Priyanto, Anugerah Dany (2014) Screening and Characterization of Angiotensin-I Converting Enzyme Inhibitory Peptides Derived from Enzymatic Hydrolysate of Bitter Melon Seed Proteins. Magister thesis, Universitas Brawijaya.

Abstract

Enzim Konversi Angiotensin (ACE) memainkan peran imperatif dalam sistem tekanan darah. Ini menghasilkan vasokonstriksi yang kuat dengan mengkonversi angiotensin-i ke angiotensin-II. Pengaturan konter ACE dapat mengurangi tekanan darah. Tujuan dari penelitian ini Areto Temukan dan ciri peptida penghambatan ACE dari biji melon pahit. Peptida penghambatan angiotensin-i converting enzim (ACE) berhasil disaring dari protein bitter melon (Mombordica Charantia) Hidrolisat dengan pencernaan termolissin. Fraksi ultrafiltrated menggunakan 3 KDA membran cut-off berat molekul dipisahkan oleh fase terbalik kromatografi cair kinerja tinggi (RP-HPLC), yang diikuti oleh urutan analisis peptida menggunakan kromatografi cair-tandem spektrometri massa (LC-MS) dan Pengujian menggunakan uji aktivitas penghambatan ACE. VY-7 dan VG-8 diidentifikasi sebagai peptida penghambatan ACE yang lebih tinggi dengan nilai IC 50 sebesar 7,98 ± 0,99 dan 13,39 ± 0,59 μm, masing-masing. Kami memiliki bukti bahwa peptida berasal dari α -mmc dan map30. Selain itu, plot Lineweaver-Burk menunjukkan bahwa peptida dengan IC 50 rendah, VY-7, bertindak sebagai penghambat kompetitif dengan konstanta disosiasi (K I) 19,28 μm. VY-7 ditemukan stabil untuk pencernaan gastrointestinal yang disimulasikan secara in vitro. Studi docking molekuler menunjukkan bahwa VY-7 secara preferensi berlabuh ke dalam situs pengikatan yang sama dengan obat komersial yang berinteraksi dengan ace somatik manusia. Dalam pose berlabuh, VY-7 berinteraksi dengan wilayah ACE S1 (GLU 384 OE2 dan ALA 354 O), wilayah S1` (Glu 162 OE2), dan wilayah S2` (353 NE2). Dalam model hewan tekanan darah tinggi, VY-7 dosis pada 2 mg / kg berat badan secara signifikan menurunkan tekanan darah sistolik pada 4 jam setelah pemberian tetapi memulihkan tekanan darah sistolik sebesar 24 jam. Vy-7 karenanya menunjukkan potensi untuk dikembangkan sebagai tekanan darah yang menurunkan nutraceutical.

English Abstract

Angiotensin-converting enzyme (ACE) plays an imperative role in the blood pressure system. It generates strong vasoconstriction by converting angiotensin-I to angiotensin-II. Counter regulation of ACE may reduce blood pressure. The aims of this study areto discover and characterize ACE inhibitory peptides from bitter melon seeds. Angiotensin-I converting enzyme (ACE) inhibitory peptides were successfully screened from bitter melon ( Momordica charantia ) seed protein hydrolysates by thermolysin digestion. The ultrafiltrated fractions using 3 kDa molecular weight cut-off membrane were separated by reverse phase high performance liquid chromatography (RP-HPLC), which was followed by sequence analysis of peptides using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and testing using an ACE inhibitory activity assay. VY-7 and VG-8 were identified as the higher ACE inhibitory peptides with IC 50 values of 7.98 ± 0.99 and 13.39 ± 0.59 μM, respectively . We have evidence that the peptides are derived from α -MMC and MAP30. In addition, Lineweaver-Burk plots indicated that the peptide with the low IC 50 ,VY-7, acts as a competitive inhibitor with a dissociation constant ( K i ) of 19.28 μM . VY-7 was found to be stable to in vitro simulated gastrointestinal digestion. Molecular docking studies showed that VY-7 preferentially docked into the same binding sites as commercial drugs that interact with human somatic ACE. In the docked pose, VY-7 interacted with the ACE S1 region (Glu 384 OE2 and Ala 354 O), S1` region (Glu 162 OE2), and S2` region (His 353 NE2). In an animal model of high blood pressure, VY-7 dosed at 2 mg/kg body weight significantly decreased the systolic blood pressure at 4 h after administration but restored the systolic blood pressure by 24 h. VY-7 hence shows potential to be developed as a blood-pressure lowering nutraceutical.

Item Type: Thesis (Magister)
Identification Number: TES/572.7/PRI/s/041404769
Subjects: 500 Natural sciences and mathematics > 572 Biochemistry > 572.7 Enzymes
Divisions: S2/S3 > Magister Teknologi Hasil Pertanian, Fakultas Teknologi Pertanian
Depositing User: Endro Setyobudi
Date Deposited: 22 Aug 2014 15:42
Last Modified: 31 Mar 2022 07:39
URI: http://repository.ub.ac.id/id/eprint/157696
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