Rifqiyah Nur Umami, Rifqiyah Nur
Research Center for Biotechnology, Indonesian Institute of Sciences, Bogor, Indonesia.

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Identification of Bioactive Compound from Microalga BTM 11 as Hepatitis C Virus RNA Helicase Inhibitor Mustopa, Apon Zaenal; Umami, Rifqiyah Nur; Putri, Prabawati Hyunita; Susilaningsih, Dwi; Farida, Hilda
JURNAL BIOLOGI INDONESIA Vol 11, No 2 (2015): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1133.514 KB) | DOI: 10.14203/jbi.v11i2.2198

Abstract

ABSTRACTHepatitis C virus (HCV) is the major causative agent of chronic liver disease. Recently, the inhibition of NS3 RNA helicase/ATPase activity is being explored as the specifically targeted antiviral therapy (STAT) against HCV infection. This study was aimed to elucidate potential candidates for anti-HCV therapy derived from Indonesian indigenous microalgae. The microalga designated as BTM 11 was isolated and cultured. Methanol extract of BTM 11 was screened as the opponent of purified HCV NS3 RNA helicase enzyme through colorimetric ATPase assay. Screening of chemical compound and fractionation by using gel filtration chromatography with eluent of methanol : chloroform (1:99) were conducted for identification and isolation of the bioactive compounds. The third fraction of fractionated sample showed a relatively strong ATPase inhibitory effect (81.23 ± 2.25 %) compared to the negative control. Further analysis of third fraction using thin layer chromatography (TLC) with eluent of chloroform : methanol (9:2) gave two spots with the Rf value of 0.8 and 0.37, respectively. In addition, high performance liquid chromatography (HPLC) analysis showed absorption peak with the highest abundance at the retention time of 12.483 and 16.617 minutes which absorbed at 266 and 230 nm wavelenght, respectively. According to those analyses, this study suggests that bioactive compounds derived from BTM 11 were classified as the groups of flavonoids and feasible as potential candidates for anti-HCV therapy through the inhibitory effect of NS3 RNA helicase/ATPase activity. Keywords: Hepatitis C Virus, NS3 RNA helicase, ATPase, Microalga, Flavonoids 
CLONING, EXPRESSION, AND PARTIAL PURIFICATION OF PLANTARICIN W LOCUS PRODUCED BY Lactobacillus plantarum S34 [Kloning, Ekspresi, dan Purifikasi Parsial Lokus Plantarisin W Diproduksi oleh Lactobacillus plantarum S34] Umami, Rifqiyah Nur; Mustopa, Apon Zaenal; Sukmarini, Linda; Danuri, Hasim; Putri, Andini Setyanti; Wibowo, Krisna Dwi Aria
BERITA BIOLOGI Vol 16, No 1 (2017)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (2760.052 KB) | DOI: 10.14203/beritabiologi.v16i1.2174

Abstract

Lactobacillus plantarum S34 dilaporkan mempunyai aktivitas antibakteri yang terkait dengan produksi bakteriosin. Bagian dari gen yang menyandikan salah satu lokus bakteriosin yang diproduksi oleh L. plantarum S34, disebut dengan plantarisin W (plnW), diamplifikasi dari plasmid dan dikloning menggunakan sistem vektor pGEM®-T Easy ke dalam Escherichia coli DH5?. Sekuens nukleotida plnW (± 405 pb) diidentifikasi sebagai protein integral membran. Lebih lanjut, plnW diekspresikan secara heterologus sebagai fusi protein dengan His(6)-tag tioredoksin menggunakan vektor ekspresi pET-32a(+) ke dalam E. coli BL21 (DE3) pLysS. Protein fusi rekombinan plnW terdapat dalam sitoplasma sel, tetapi selain fraksi terlarut terdapat juga fraksi tidak terlarut berupa badan inklusi. Purifikasi parsial dilakukan menggunakan kromatografi afinitas ligan Co2+ untuk fraksi terlarut dan metode elektroelusi gel poliakrilamid untuk fraksi tidak terlarut. Massa molekul berukuran kurang lebih 33 kDa terdeteksi berdasarkan pemisahan SDS-PAGE dan dikonfirmasi dengan Western blot sebagai protein fusi rekombinan plnW. Protein yang sudah terpurifikasi bermanfaat untuk mengetahui kaitan antara struktur dan fungsi bakteriosin.
IDENTIFICATION OF BIOACTIVE COMPOUND FROM MICROALGA BTM 11 AS HEPATITIS C VIRUS RNA HELICASE INHIBITOR Mustopa, Apon Zaenal; Umami, Rifqiyah Nur; Putri, Prabawati Hyunita; Susilaningsih, Dwi; Farida, Hilda
JURNAL BIOLOGI INDONESIA Vol 11, No 2 (2015): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v11i2.2198

Abstract

ABSTRACTHepatitis C virus (HCV) is the major causative agent of chronic liver disease. Recently, the inhibition of NS3 RNA helicase/ATPase activity is being explored as the specifically targeted antiviral therapy (STAT) against HCV infection. This study was aimed to elucidate potential candidates for anti-HCV therapy derived from Indonesian indigenous microalgae. The microalga designated as BTM 11 was isolated and cultured. Methanol extract of BTM 11 was screened as the opponent of purified HCV NS3 RNA helicase enzyme through colorimetric ATPase assay. Screening of chemical compound and fractionation by using gel filtration chromatography with eluent of methanol : chloroform (1:99) were conducted for identification and isolation of the bioactive compounds. The third fraction of fractionated sample showed a relatively strong ATPase inhibitory effect (81.23 ± 2.25 %) compared to the negative control. Further analysis of third fraction using thin layer chromatography (TLC) with eluent of chloroform : methanol (9:2) gave two spots with the Rf value of 0.8 and 0.37, respectively. In addition, high performance liquid chromatography (HPLC) analysis showed absorption peak with the highest abundance at the retention time of 12.483 and 16.617 minutes which absorbed at 266 and 230 nm wavelenght, respectively. According to those analyses, this study suggests that bioactive compounds derived from BTM 11 were classified as the groups of flavonoids and feasible as potential candidates for anti-HCV therapy through the inhibitory effect of NS3 RNA helicase/ATPase activity. Keywords: Hepatitis C Virus, NS3 RNA helicase, ATPase, Microalga, Flavonoids