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All Journal HAYATI Journal of Biosciences Jurnal Pengolahan Hasil Perikanan Indonesia ILMU KELAUTAN: Indonesian Journal of Marine Sciences Indonesian Journal of Cancer Chemoprevention BERITA BIOLOGI JURNAL BIOLOGI INDONESIA ANNALES BOGORIENSES Annales Bogorienses
Apon Zaenal Mustopa
Research Center for Biotechnology, Indonesian Institute of Sciences Jalan Raya Bogor km 46, Cibinong 16911, Bogor, Indonesia
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Earth & Planetary Sciences Immunology & microbiology Medicine & Pharmacology

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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 
APLIKASI MARKA MOLEKULER PADA BUAH DAN BIJI KOPI ASAL KALIMANTAN TIMUR [Molecular marker application of cherry and green bean of East Kalimantan coffee] Fatimah, Fatimah; Urnemi, Urnemi; Mustopa, Apon Zaenal; Syahrumsyah, Hudaida
BERITA BIOLOGI Vol 13, No 1 (2014)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (133.562 KB) | DOI: 10.14203/beritabiologi.v13i1.649

Abstract

Two quantitative traits, cherry and green bean characters are the important phenotypic selection in coffee breeding practice. The important well-known character from coffee markets is cherry and bean size. In this study, 43 genotypes of coffee were collected from four districts in East Kalimantan i.e. Kutai Kertanegara, Kutai Timur, Berau dan Paser Utara. The objective of this study was to identify cherry and green bean character using quantitative trait locus (QTL) molecular marker, genetic variation from developed alleles, cluster analysis and association analysis of molecular marker, and phenotype observation. Based on polymorphic information content (PIC) of primers used in this study, the genetic variation was low. Based on cluster analysis, two major groups were identified. The first group corresponds to Arabika that consisted of 3 districts, Kutai Timur, Berau and Paser Utara. The second group correspond to Robusta mostly from Kutai Kertanegara.Significant association of primer markers M480 and M312 with QTL has suggested that they can be used as specific primers linked to size of cherry and green bean.Furthermore,they were potential marker assisted breeding in coffee breeding program.
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.
Enterococcus faecium 1.15 Isolated from Bakasam Showed Milk Clotting Activity Putranto, Wendry Setiyadi; Suradi, Kusmajadi; Chairunnisa, Hartati; Mustopa, Apon Zaenal; Giriwono, Puspo Edi; Kusumaningrum, Harsi Dewantari; Suhartono, Maggy Thenawidjaja
ANNALES BOGORIENSES Vol 21, No 1 (2017): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (499.913 KB) | DOI: 10.14203/ab.v21i1.293

Abstract

The Lactic Acid Bacteria with Milk Clotting Activity (MCA) were isolated from Bakasam, an Indonesian traditional fermented meat. The isolate screening was carried out using modified method of Skim Milk Agar and Milk Clotting Activity Test, and the isolate was then identified using 16S rRNA. We found 4 isolates that showed MCA of 18-20 SU/ml. Identification using 16S rRNA indicated that the isolate ALG.1.15 was 99% (FR3-F primer) and 99% (FR3-R primer) identic with Enterococcus faecium. The isolate potentially produced renin-like protease to subtitute renin from veal.  
Molecular Identification of Microalgae BTM 11 and its Lectin Isolation, Characterization, and Inhibition Activity Mustopa, Apon Zaenal; Isworo, Rhestu; Nurilmala, Mala; Susilaningsih, Dwi
ANNALES BOGORIENSES Vol 20, No 2 (2016): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ab.v20i2.251

Abstract

BTM 11 is unknown species of microalgae, but has active compounds that can inhibit viruses. One of proteins produced by microalgae is a lectin. Lectin is a carbohydrate-binding protein found in various microalgae that show antiviral and antibacterial activity. The purpose of this study was to perform identification of the species of microalgae BTM 11, isolation, characterization, and assay of lectin inhibitory activity. Microalgae BTM 11 was identified to have homology with Cyanobacterium 99% and Geitlerinema sp 98%. Lectin of microalgae BTM 11 was isolated by ammonium sulfate precipitation of 75% with a molecular weight of 17 kDa. Lectin protein activity of microalgae BTM 11 was able to inhibit the enzyme activity of RNA helicase hepatitis C by 57.90% and 27.55%. In addition, the protein was able to suppress the activity of Staphylococcus aureus ATCC 6538, E. coli EPEC K.1.1. and Salmonella typhii ATCC 25241. Activitiy of lectin was stable at 30 °C and was unaffected by the action of the enzyme. These results indicate that lectin of microalgae BTM 11 could be a alternative to antiviral and antibacterial proteins.
Pengaruh Suhu, pH, Enzim dan Surfaktan terhadap Plantarisin F Rekombinan Enkapsulasi sebagai Antibakteri Staphylococcus aureus dan Salmonella typhi Mustopa, Apon Zaenal; Hasim, Hasim; Amelia, Suci
JURNAL BIOLOGI INDONESIA Vol 14, No 1 (2018): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v14i1.3664

Abstract

ABSTRACTStaphylococcus aureus causes diarrhea, which is one of the main cause of infant mortality, where asSalmonella typhi causes typhoid fever with the incidence rate of 180/100,000/year. Plantarisin F is anantimicrobial peptide that can inhibit the growth of S. aureus and S. typhi. The aim of this study is to determinethe effect of temperature, pH, enzymes, and surfactants of encapsulated F recombinant plantarisin. Plantarisin F(1.61%) encapsulated with maltodextrin (5.36%) and skim milk (2.68%) using the spray dry with inlettemperature 150ºC produced particles that are generally spherical with a rough texture, range in size, yield25.03%, and had good antibacterial activity against S. aureus and S. typhi. The antibakterial activity plantarisinF encapsulated is not affected by the treatment temperature (40ºC-100ºC), pH (2-12), enzyme (proteinase-K,catalase, lysozyme, pepsin, trypsin), dan surfaktan (SDS, urea, triton X-100, PMSF, EDTA) treatment.Keywords: Antibacterial, Encapsulation, Plantarisin F, Salmonella typhi, Staphylococcus aureus
Optimization of Expression Condition, Two Dimensional And Melting Point-Based Characterization of Recombinant Human Interferon Alpha-2a Fusion and Non Fusion Forms Ningrum, Ratih Asmana; Wardhani, Widdya Kusuma; Wahyuni, Ike; Mustopa, Apon Zaenal
ANNALES BOGORIENSES Vol 22, No 2 (2018): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ann.bogor.2018.v22.n2.57-64

Abstract

     Recombinant Human Interferon Alpha-2a (rhIFNα-2a) is a therapeutic protein that used in hepatitis and cancer treatments. In our previous research, we developed higher molecular weight of the protein through human serum albumin fusion. The fusion and non fusion form of rhIFNα-2a were produced in Pichia pastoriswith 86 kDa and 19 kDa in size respectively. In previous research, protein yield was not reproducible due to unoptimized expression conditions. This reseach was aimed to optimize expression condition process and to characterize the fusion and non fusion forms of rhIFNα-2a. The parameters to observe in overproduction include nutrient (media and methanol concentration) and non nutrient (temperature andincubation period). Affinity and size exclusion cromatographicwere compared in protein purification. BCA assay was used to determine quantity of protein. Protein characterization was conducted using two-dimensional SDS PAGE and denaturation analyses. The optimal condition of expression was achieved using complex media with 1% of methanol for 3 day incubation period at 25°C. The protein yield was reproducible and higher comparing to previous research. Affinity chromatography resulted in higher purity of the proteins comparing to size exclusions. Characterization using two dimensional gel analysis revealed that isoelectric point of rhIFNα-2a is 6.5 for fusion form and 6.0 for non fusion form. The melting points of fusion protein were 56°C and 62°C whilst that of non fusion was 56°C.
Partial Purification, Characterization, and Application of Extracellular Aspartic Protease from Lactobacillus casei WSP in Producing the Bioactive Peptides with Antibacterial and Antioxidant Activity Solikhin, Akhmad; Mustopa, Apon Zaenal; Suharsono, Suharsono; Putranto, Wendry Setiyadi
ANNALES BOGORIENSES Vol 22, No 2 (2018): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ann.bogor.2018.v22.n2.47-56

Abstract

   Lactobacillus casei WSP-derived an aspartic protease was sequentially purified by using chromatography gel filtration sephadex G-50. It resulted in a 22.81-fold increase of specific activity (51.5 U/mg) with a final yield of 1.9%. The estimated molecular weight of the purified enzyme was 37 kDa and showed gelatinolytic activity in zymogram assay. The enzyme exhibited optimum activity at 40ºC and pH 6 with casein as the substrate. Enzyme activity was significantly inhibited by pepstatin A (0.5 mM and 1 mM), confirming that this enzyme is a group of aspartic proteases, while other inhibitors such as EDTA, PMSF and iodoacetic acid showed no inhibition effect on the activity of enzyme. The addition of metal ion to the enzyme decreased enzyme activity, indicating the proteolytic enzyme was metal ion- dependent. Denaturant such as DDT tended to increase caseinolytic activity. Furthermore, this enzyme was capable of generating the new peptides from skimmed milk with the size 8 kDa, 10 kDa and 15 kDa. These peptides have potential as antibacterial and antioxidant agents.
AKTIVITAS ANTIBAKTERI DAN ANTIOKSIDAN PEPTIDA SUSU KAMBING HASIL HIDROLISIS DENGAN PROTEASE LACTOBACILLUS PLANTARUM S31 Herlina, Nina; Mustopa, Apon Zaenal; Surachma, Rahma Sari; Triratna, Lita; Kartina, Gina; Alfisyahrin, Wida Nurul
JURNAL BIOLOGI INDONESIA Vol 15, No 1 (2019): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v15i1.3762

Abstract

ABSTRACTResearch on the hydrolysis of milk with various protease obtained from the digestive tract, plants and animals has been widely carried out, but the hydrolysis process of goat milk with protease enzymes from Lactobacillus plantarum S31 isolated from bekasam is still widely unknown. This study aims to determine the antibacterial and antioxidant activity of peptides hydrolyzed by protease enzymes from L. plantarum S31. Isolation and purification of extracellular proteases was carried out with 80% ammonium sulfate saturation, dialysis and G50 Sephadex matrix. The results of hydrolysis showed that fraction 11 has the highest antioxidant activity. Goat milk with pH 5 has a greater inhibitory activity of about 29%, which is 5% compared to goat milk pH 7. This fraction also has a quite good antibacterial activity of Entero Phatogenic Escherichia coli (EPEC K1.1) bacteria, Staphylococcus aureus and Listeria monocytogenes.  Keywords: goat milk, hydrolysis, antibacterial, antioxidant, Lactobacillus plantarum
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 
Co-Authors Ai Hertati, Ai Akhmad Solikhin Akmaliyah, Rizna Alfisyahrin, Wida Nurul Amani, Febriyanti Nur Amelia, Suci Amelia, Suci Andi Asmawati Azis Anika Prastyowati, Anika Anja Meryandini Anna Ida Sunaryo Purwiyanto Arifah, Rosyida Khusniatul Ario Betha Juanssilfero Chairunnisa, Sheila Chihombori, Tatenda Calvin DWI SUSILANINGSIH Ekawati, Nurlaili Eva Johannes Farida, Hilda Farida, Hilda Fatimah fatimah Fatimah Fidien, Khadijah Alliya Fidien Firdaus, Moh Egy Rahman GINA KARTINA Harsi D. Kusumaningrum Hartati Chairunnisa Hartoni Hasim - Hasim Hasim Huda Shalahudin Darusman Hudaida Syahrumsyah, Hudaida Ikramullah, Muh. Chaeril Iman Rusmana Irawan, Herman Irawan, Shasmita Isworo, Rhestu Isworo, Rhestu Jendri Mamangkey Joko Wahyono, Daniel Kartika Sari Dewi Kusdianawati Kusmajadi Suradi Kustiariyah Tarman Kusumawati, Arizah LINDA SUKMARINI Lita Triratna Magdalena Litaay Maggy Thenawidjaja Suhartono Mala Nurilmala Manguntungi, Baso Maulidiani, Maulidiani Meilina, Lita Meiyerani, Jeni Melki Mustafawi, Wike Zahra Mustika Tuwo, Mustika Mutiara, Ilma Nina Herlina Novianti, Ela Nur Amani, Febriyanti Nur Umami, Rifqiyah Nurfatwa, Maritsa Pandayu, Iga Firmansyah Popi Hadi Wisnuwardhani, Popi Hadi Puspo Edi Giriwono Putri, Andini Setyanti Putri, Prabawati Hyunita Putri, Prabawati Hyunita Ramadhian, M. Zalfa RATIH ASMANA NINGRUM Ratna, Lita Tri Refli, Redoyan Rifqiyah Nur Umami, Rifqiyah Nur Rika Indri Astuti Saputri, Dinar Suksmayu Saputri Sofyana, Neng Tanty Sri Swasthikawati, Sri Suharli, Lili Suharsono Suharsono Surachma, Rahma Sari Tri Wibawa Trinugroho, Joko P Urnemi Urnemi, Urnemi Vanggi, Leggina Rezzy Vanggy, Leggina Rezzy Wahyuni, Ike Wahyuni, Ike Wardhani, Widdya Kusuma Wardhani, Widdya Kusuma Wendry Setiyadi Putranto Wibowo, Krisna Dwi Aria Wijaya, Hans Wike Ayu Eka Putri Yudi Purnomo