isolasi dan purifikasi inhibitor rna helikase virus

14
ABSTRAK
SKOTIA FITRIASTRI PUTRI. Isolasi dan Purifikasi Inhibitor RNA Helikase
Virus Hepatitis C dari Bakteriosin Bakteri Asam Laktat S34. Dibimbing oleh
SURYANI dan A. ZAENAL MUSTOPA.
Terapi target molekular merupakan salah satu penanggulangan penyakit
virus hepatitis C yang sedang dikembangkan. Penghambatan NS3 helikase atau
RNA helikase virus hepatitis C dengan peptida inhibitor merupakan salah satu
terapi target molekuler. Bakteriosin merupakan salah satu peptida inhibitor yang
memiliki kemampuan untuk menghambat virus hepatitis C melalui mekanisme
penghambatan NS3 helikase. Bakteriosin adalah peptida yang dihasilkan bakteri
asam laktat. Tujuan penelitian ini untuk melakukan isolasi dan purifikasi
bakteriosin dari isolat S34 yang memiliki aktivitas yang tinggi dalam menghambat
kerja RNA helikase virus hepatitis C. Bakteri asam laktat S34 yang diidentifikasi
dengan metode amplifikasi 16S rRNA dan sekuensing DNA merupakan bakteri
Lactobacillus plantarum WCFS1. Pertumbuhan bakteri tersebut mencapai fase
logaritma pada jam ke-7 dan fase stasioner jam ke-10. Bakteriosin diisolasi dan
dipurifikasi melalui presipitasi amonium sulfat dengan kejenuhan 80% dan
kromatografi gel filtrasi serta dilakukan karakterisasinya meliputi analisis bobot
molekul dan pengukuran kadar protein. Bakteriosin dari bakteri asam laktat S34
memiliki persentase inhibisi terhadap RNA helikase virus hepatitis C sebesar
64.20%, diduga bakteriosin kelas 2, bersifat termostabil, memiliki bobot molekul
pada kisaran 1.7-4.6 kDa dengan aktivitas spesifik sebesar 2.84 x 106 U/mg.
15
ABSTRACT
SKOTIA FITRIASTRI PUTRI. Isolation and Purification RNA Helicase Inhibitor
of Hepatitis C Virus From S34 Lactic Acid Bacteria Bacteriocin. Under the
direction of SURYANI and A. ZAENAL MUSTOPA.
Target molecular therapy is one of method to prevent hepatitis C virus
which was developed recently. NS3 Helicase/RNA helicase hepatitis C virus
inhibition using inhibitory peptide are target in molecular therapy. Bacteriocin are
inhibitory peptides which can inhibit hepatitis C virus through inhibition of NS3
helicase. Bacteriocin is a peptide which is produced by lactic acid bacteria. The
aim of this research were to isolate and to purify bacteriocin from S34 lactic acid
bacteria which has high activity to inhibit RNA helicase of hepatitis C virus. S34
lactic acid bacteria were identified using 16S RNA amplification and DNA
sequencing identification of S34 lactic acid bacteria was Lactobacillus plantarum
WCFS1. The bacterial growth reached logaritmic phase at 7 hours and stationary
phase at 10 hours. Bacteriocin were isolated and purified by ammonium sulphate
precipitation with 80% saturation, gel filtration chromatography, and
characterization of bacteriocin through molecular weight analysis and measured
protein concentration. Bacteriocin from S34 lactic acid bacteria were included in
2nd class bacteriocin, had 64.20 % of inhibition against RNA helicase hepatitis C
virus, had molecular weight about 1.7-4.2 kDa, and specific activity in the amount
of 2.84 x 106 U/mg.