Sintesis Tetrapeptida FmeLAP (Phe-meLeu-Ala-Pro) sebagai Kandidat Antioksidan dengan Metode Solid Phase Peptide Synthesis
Abstract
ABSTRACT: Antioxidant peptides are a group of peptides that can capture free radicals, so as to prevent premature aging and degenerative diseases, one of the natural antioxidant peptide compounds that has been found by previous researchers is the linear tetrapeptide compound FLAP (Phe-Leu-Ala-Pro) isolated from the sea cucumber Acaudina Molpadiodes found in Zheijing China and reported to have poor antioxidant activity. In this study, tetrapeptide FmeLAP (Phe-meLeu-Ala-Pro) has been synthesized using the Solid phase peptide synthesis method on 2-chlorotrityl chloride resin. Fmoc was used as a protective amino acid group. In the preparation of the peptide fragment, a combination of HOBt and HBTU was used as a coupling reagent in the synthesis of FmeLAP. After the peptide fragments were arranged on the resin, the combination of trifluoroacetic acid in dichloromethane was added for the process of releasing the peptides from the resin. Concentration was carried out using a rotary evaporator. Then the characterization was carried out using a mass spectrophotometer, the peak of the fragment was obtained at m/z 461 and a purity test was carried out using RP-HPLC the results were obtained at 11.035 minutes. In testing the antioxidant activity using the DPPH method, the results obtained from the tetrapeptide compound (Phe-meLeu-Ala-Pro) is an inhibition value of 47% which indicates that substitution of leucine to methyl leucine did not improve the antioxidant activity of the compound.
Keywords: Antioxidant, FmeLAP tetrapeptide, Solid Phase Peptide Synthesis (SPPS)
ABSTRAK: Peptide antioksidan merupakan kelompok peptide yang dapat menangkap radikal bebas, sehingga dapat mencegah penuaan dini dan penyakit degenerative, salah satu senyawa peptide antioksidan alami yang telah ditemukan peneliti sebelumnya yaitu senyawa tetrapeptida linier FLAP (Phe-Leu-Ala-Pro) yang diisolasi dari teripang Acaudina Molpadiodes yang di temukan di Zheijing China dan dilaporkan memiliki aktifitas antioksidan yang kurang baik. Telah dilakukan sintesis Tetrapeptida FmeLAP (Phe-meLeu-Ala-Pro) dengan menggunakan metode Solid phase peptide synthesis pada resin 2-klorotritil klorida. Pada penelitian ini digunakannya Fmoc sebagai gugus pelindung asam amino. Pada penyusunan fragmen peptida digunakan kombinasi HOBt dan HBTU sebagai reagen pengkopling dalam sintesis FmeLAP. Setelah fragmen peptida tersusun pada resin, ditambahkan kombinasi asam triflouroasetat dalam diklorometana untuk proses pelepasan peptida dari resin. Dilakukan pemekatan menggunakan rotary evaporator. Kemudian dilakukan karakterisasi menggunakan spektrofotometer massa didapat puncak fragmennya pada m/z 461 dan dilakukan uji kemurnian menggunakan RP-HPLC hasil yang didapat pada menit ke 11,035. Pada pengujian aktivitas antioksidan dengan menggunakan metode DPPH, hasil yang didapat dari senyawa tetrapeptida (Phe-meLeu-Ala-Pro) yaitu nilai inhibisi sebesar 47% yang menunjukan bahwa pergantian asam amino leusin menjadi metil leusin tidak meningkatkan aktivitas antioksidan dari senyawa tetrapeptida.
Kata Kunci: Antioksidan, tetrapeptida FmeLAP, Solid Phase Peptide Synthesis (SPPS)
Keywords
Full Text:
PDFReferences
Bougatef, A., Nedjar-Arroume, N., Manni, L., Ravallec, R., Barkia, A., Guillochon, D., dkk. (2010). Pemurnian dan identifikasi peptida antioksidan baru dari hidrolisat enzimatik dari protein produk samping sardinelle (Sardinella aurita). Kimia Pangan. 118, 559–565.
Byun, H.G., Lee, J.K., Park, H.G., Jeon, J. K. & Kim, S.K. (2009). Antioxidant peptides isolated from the marine rotifer, brachionus rotundiformis. Process Biochemistry. 44: 842846.
Chan, W.C. & White, P.D. (2000). Fmoc Solid Phase Peptide Synthesis: A Practical Approach. Oxford University Press. New York.
Dehpour AA. Ebrahimzadeh MA. Fazel NS dan Mohammad NS. (2009). Antioxidant Activity of methanol extract of ferula assafoetida and it's essential oil composition. Grasas Aceites. 60: (4). 405-412.
Elias, R.J., Kellerby, S.S., & Decker, E.A. (2008). Antioxidant Activity Of Proteins and Peptides. Critical Reviews in Food Science and Nutrition. 48(5): 430441.
Goldberg, G. (2003). Diet and Health. I Owa State Press. Blackwell Publishing Company, 2121 State Avenue, Ames. USA.
Hernani dan Rahardjo, M. (2005). Tanaman Berkhasiat Antioksidan. Penebar Swadaya. Jakarta; hal 1-20; 62-63.
Jeong, J. B., Jeong, H. J., Park, J. H., Lee, S. H., Lee, J. R., Lee, H. K., et al. (2007). Cancer- preventive peptide lunasin from Solanum nigrum L. inhibits acetylation of core histones H3 and H4 and phosphorylation of retinoblastoma protein (Rb). Journal of Agricultural Food Chemistry, 55, 10707−10713.
Jin, H. X., Xu, H. P., Li, Y., Zhang, Q. W., & Xie, H. (2009). Preparation and evaluation of peptides with potential antiokxidant activity by microwave assisted enzymatic hydrolysis of collagen from sea cucumber acaudina molpadioides obtained from Zhejiang Province in China. Marine drugs, 17(3), 169.
Kim, E., Lee, S., Jeon, B., Moon, S., Kim, B., Park, T., Park, P. (2009). Purification and characterisation of antioxidative peptides from enzymatic hydrolysates of venison protein. Food Chemistry, 114 (4), 1365–1370.
Maharani, R., Sumiarsa, D., Marpaung, C., Zainuddin, A., Hidayat, A. T., Harneti, D., Nurlelasari, Supratman, U. (2019). Sintesis Tetrapeptida PADY menggunakan Metode Fasa Padat dan Aktivitas Antioksidannya. Jurnal Kimia Valensi. Bandung: Departemen Kimia, Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Padjadjaran
Minervini, F. F. A., Rizzello, C. G., Fox, P. F., Monnet, V., & Gobbetti, M. (2003). Angiotensin I-converting-enzyme-inhibitory and antibacterial peptides from Lactobacillus helveticus PR4 proteinase-hydrolyzed caseins of milk from six species. Applied and Environmental Microbiology, 69, 5297-5305.
Molyneux, P. (2004). The Use of The Stable Free Radical Diphenylpicrylhydrazyl (DPPH) For Estimating Antioxidant Activity. Songklanakarin Journal of Science and Technology, 26 (2): 211-219.
Murray, R.K. (2003). Biokimia Haper. Dalam: Anna P. Bani. Edisi ke-25. Jakarta, EGC.
Sitompul, S. (2004). Analisis Asam Amino dalam Tepung Ikan dan Bungkil. Kedela. Buletin Teknik Pertanian. Vol 9. Nomor 1.
Sloane, Ethel. (2004). Anatomi dan Fisiologi Untuk Pemula. Jakarta: Penerbit Buku Kedokteran EGC.
Wildman, R.E.C. (2001). Handbook of Nutrateutical and Founctional Foods. CRC Press. USA.
Fauzi, Nur Muhammad. (2021). Uji Kualitatif dan Uji Aktivitas Antioksidan Ekstrak Etanolik Buah Maja (Aegle Marmelos (L.)Correa) dengan Metode DPPH. Jurnal Riset Farmasi, 1(1), 1-8.
DOI: http://dx.doi.org/10.29313/.v0i0.30349
