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 
Cloning, Expression, and Bioinformatics Modeling of Human Papillomavirus Type 52 L1/L2 Chimeric Protein in Escherichia coli BL21 (DE3) Ikramullah, Muh. Chaeril; Mustopa, Apon Zaenal; Wibawa, Tri; Hertati, Ai; Umami, Rifqiyah Nur; Ratna, Lita Tri; Irawan, Shasmita; Firdaus, Moh Egy Rahman; Darusman, Huda Salahudin
HAYATI Journal of Biosciences Vol. 31 No. 5 (2024): September 2024
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.31.5.891-902

Abstract

Human papillomavirus (HPV) L1 major capsid protein generates a highly immunogenic virus like particles (VLPs), which have been used as the main component of its prophylactic vaccine. However, the neutralizing antibodies against L1 VLPs are mostly type specific and may not be effective to prevent infection from different strains of HPV. On the other hand, HPV L2 minor capsid protein has low antigenic variation, thus can induce cross-neutralization. This study aims to obtain HPV 52 L1/L2 chimeric protein, which is designed based on HPV type 52 as one of the most circulated high-risk types in Indonesia, to develop a broad-spectrum HPV vaccine. Substitution of HPV 52 H4 helix L1 region with an HPV 52 L2 epitope was carried out using overlap extension PCR. HPV 52 L1/L2 chimeric gene was constructed into pET-SUMO expression vector and expressed in Escherichia coli BL21 (DE3). Bioinformatics modeling suggested that L2 epitope was located inside of the loop region in monomer form, and on the contrary, it was located outside of the pentamer surface. Furthermore, B cell and T cell epitopes predictions were conducted using Immune Epitope Database (IEDB) analysis. The B cell epitopes prediction revealed eleven potential epitopes, whereas the T cell epitopes prediction showed seven potential epitopes for each MHC class I and MHC class II. This study showed that HPV 52 L1/L2 chimeric protein has the potential to induce cross-neutralizing antibodies and can be developed as a promising candidate for a new HPV vaccine.
Potential Probiotic Yeasts of the Pichia Genus Isolated from ‘Dadih’, a Traditional Fermented Food of West Sumatra, Indonesia Chihombori, Tatenda Calvin; Mustopa, Apon Zaenal; Astuti, Rika Indri; Mutiara, Ilma; Refli, Redoyan; Umami, Rifqiyah Nur; Fatimah; Irawan, Herman; Ekawati, Nurlaili; Trinugroho, Joko P; Akmaliyah, Rizna; Chairunnisa, Sheila; Amani, Febriyanti Nur; Manguntungi, Baso; Hertati, Ai; Mamangkey, Jendri
HAYATI Journal of Biosciences Vol. 32 No. 2 (2025): March 2025
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.32.2.320-340

Abstract

Fermented buffalo milk, known as dadih, serves as a reservoir of potential probiotic yeasts. Over the past two decades, probiotic yeasts have gained increasing attention in both basic and clinical sciences due to their health benefits. This study aimed to isolate and characterize probiotic yeasts from dadih. Yeasts were isolated using yeast Extract, peptone, and dextrose (YPD) medium, and molecularly identified through 18S-rRNA sequencing. Probiotic potential was assessed by evaluating resistance to acidic pH, bile salts, proteolytic, lipolytic, and hemolytic activities. Secondary metabolites produced during fermentation were tested for antimicrobial properties. GBT30 and GBT37 isolates were selected based on their superior performance in probiotic property assays for further analysis. Molecular identification revealed these isolates as Pichia occidentalis (GBT30) and Pichia kudriavzevii (GBT37). Both strains demonstrated in vitro survivability under simulated gastrointestinal conditions and exhibited antimicrobial activity. Whole-genome sequencing of P. kudriavzevii GBT37 identified a genome size of 10,906,850 base pairs, distributed across four chromosomes with a GC content of 38.26%. Notably, secondary metabolite biosynthesis genes were located on contig 7. In addition, 26 probiotic-related genes, including GSY1, HSC82, HSP104, TPS1, ARN1, FLO1, ALA1, SIR2, and others, were identified in P. kudriavzevii GBT37, indicating its potential as a probiotic yeast. The traditional fermentation process of dadih offers probiotic yeasts with promising health benefits, supporting its potential as a functional food.