Arif Nurkanto
Research Center for Biology, Indonesian Institute of Sciences, Jalan Raya Bogor 46, Cibinong, Indonesia

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3-ACETYL -2,5,7-TRIHYDROXY-1,4-NAPHTALENEDIONE, AN ANTIMICROBIAL METABOLITE FROM THE CULTURE OF ENDOPHYTIC FUNGUS COELOMYCETES TCBP4 FROM Tinospora crispa Praptiwi, Praptiwi; Jamal, Yuliasri; Fathoni, Ahmad; Nurkanto, Arif; Agusta, Andria
Media Penelitian dan Pengembangan Kesehatan Vol 23, No 3 Sep (2013)
Publisher : Badan Penelitian dan Pengembangan Kesehatan

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (312.471 KB)

Abstract

Abstract Isolation, identification and testing of antimicrobial activity of secondary metabolites of endophytic fungal culture TCBP4 isolated from bratawali (Tinospora crispa) has been performed. The fungus TCBP4 was cultivated in Potato Dextrose Broth (PDB) for 1 month, media and fungi extracted with ethyl acetate. The extract was separated and purified by several chromatographic techniques, from which 9 fractions were obtained. Fraction 3e was purified again and was obtained 6 fractions (3e1-3e6). Fractions 3e3-3e6 were tested against bacteria isolates Staphylococcus aureus Bacillus subtilis,Eschericia coli and yeast isolate Candida albicans by microdillution method. Antimicrobial activity test result showed that fractions 3e3-3e5 had better antibacterial activity compared to chloramphenicol as commercial antibiotic. It was indicated by MIC value of the fractions was lower (8 ug/ml) compared with the antibiotic chloramphenicol (16 ug/ml). Fraction 3e3 had better antifungal activity compared to commercial antifungal nystatin and cabisidin against C. albicans. GC-MS analysis showed that the chemical constituent of 3e5 fraction was identified as 3-acetyl -2,5,7-trihydroxy-1,4-naphtalenedione . Key words : Tinospora crispa, endophytic fungi, isolation, identification, antimicrobial Abstrak Isolasi, identifikasi dan uji aktivitas antimikrobia terhadap metabolit sekunder dari kultur jamur endofit TCBP4 yang diisolasi dari tumbuhan bratawali (Tinospora crispa). Jamur endofit TCBP4 dikultivasi pada media Potato Dextrose Broth (PDB) selama 1 bulan, selanjutnya media dan jamur diekstrak dengan pelarut etil asetat. Ekstrak dipisahkan dan dimurnikan dengan beberapa teknik kromatografi sehingga diperoleh 9 fraksi. Fraksi 3e dimurnikan kembali dan diperoleh 6 fraksi (3e1-3e6). Fraksi 3e3-3e6 diuji aktivitas antibakterinya terhadap beberapa isolate bakteri Staphylo-coccus aureus Bacillus subtilis, Eschericia coli dan isolate khamir Candida albicans dengan metode mikrodilusi. Hasil uji antimikrobia menunjukkan bahwa fraksi 3e3-3e5 mempunyai aktivitas antibakteri lebih baik bila dibandingkan dengan chloramfenikol, dimana nilai MIC dari fraksi (8 ug/ml) lebih rendah dari antibiotika chloramfenikol (16 ug/ml). Fraksi 3e3 mempunyai aktivitas antijamur lebih baik bila dibandingkan dengan antijamur komersial nistatin dan kabisidin terhadap  C. albicans. Analisis GC-MS menunjukkan bahwa komponen kimia dari fraksi 3e5 diidentifikasi sebagai 3-acetyl -2,5,7-trihydroxy-1,4-naphtalenedione . Kata kunci : Tinospora crispa, jamur endofit, isolasi, identifikasi, antimikrobia
Eksplorasi Keanekaragaman Aktinomisetes Tanah Ternate Sebagai Sumber Antibiotik Nurkanto, Arif; Listyaningsih, Febrianti; Julistiono, Heddy; Agusta, Andria
JURNAL BIOLOGI INDONESIA Vol 6, No 3 (2010): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (274.268 KB) | DOI: 10.14203/jbi.v6i3.3141

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ABSTRACTExploration of Soil Actinomycetes Diversity from Ternate as Indigenous Antibiotic Sources.Actinomycetes of soil samples from Ternate, North Moluccas were isolated using SDS-YEmethod in humic acid vitamin agar. Ternate has high abundance of Actinomycetes, approximately6.00 – 487 x 104 CFU/ g soil, depends on habitat types. We have selected 60 isolates andconducted antibiotic screening against pathogenic bacteria and fungi using agar diffusionmethod and found both narrow and broad antibiotic spectrum types . Based on 16S rDNAanalysis, all Actinomycetes with antibiotic activities are belong to the genus Streptomyces. .Minimum Inhibitor Concentration (MIC) value was determined by broth microdilution method.It was found that MIC values varied, depended on microbial tested. We found two isolateswith higher antibiotic activity compared to the commercial antibiotics (chloramphenicol,erythromycin for antibacterial and nystatin, kabicidin for antifungal). Cell destruction analysiscaused antibiotic activities was conducted through leak of protein and nuclatic acid.Key words : Actinomycetes, soil, Ternate, antibiotic, cell distruction
Identifikasi Molekular dan Karakterisasi Morfo-Fisiologi Actinomycetes Penghasil Senyawa Antimikroba Nurkanto, Arif; Agusta, Andria
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.v11i2.2193

Abstract

ABSTRACTThe objectives of study were to identify antimicrobial producing Actinomycetes using 16S rDNA analyses and morphology and physiology characteristics. Eight Actinomycetes strain with the higest antibacterial and antifungal activity were selected and identified using six primers (20F, 520F, 920F, 1500R, 920R, and 520R). Morphological observation and physiology analyses were performed to the selected strain to accurately identify the strains. Morphological characters observed were aerial mycelium, spore chain, colony form, and pigment production. Physiological characterizations were antimicrobial properties, growth temperature, pH tolerance, salinity concentration for growth, sugars assimilation, and some enzymes production (arginine dihydrolase, urease, ß-glucosidase, protease, ß-galactosidase). Based on homology search by BLAST program and phylogenetic tree analyses, all of isolates were identified as the genus Streptomyces. They belong to eight different spesies. Isolates RC-SS-37-4, RC-SS-37-16 and BL-22-3 have been identified as Streptomyces costaricanus (100 %), Streptomyces costaricanus (99.8 %) and Streptomyces parvulus (98.6 %), respectively. Five isolates were identified as Streptomyces spp. (BL-36-1, BL-20-2, BL-14-2, BL-22-1 and BL-06-5) and can be presumed as new species because of the low homology value to their closest related spesies. Keywords : actinomycetes, antimicrobial, morphology, phylogenetic, physiology, 16S rRNA gene. 
Kelimpahan dan Diversitas Aktinomisetes Tanah Taman Nasional Gunung Ciremai Jawa Barat Nurkanto, Arif
JURNAL BIOLOGI INDONESIA Vol 5, No 3 (2009): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (160.878 KB) | DOI: 10.14203/jbi.v5i3.3179

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ABSTRACTThe Abundance and Diversity of Soil Actinomycetes from Ciremai National Park West Java.Actinomycetes of soil samples from Mount Ciremai National Park, West Java were been isolatedby Sodium Dodecyl Sulphate Yeast Extract (SDS YE) method in Humic Acid Vitamin Agar(HVA) medium. The colonies of actinomycetes were counted based on Total Plate Count(TPC). Diversity of actinomycetes were identified based on spore, chain spore, pigmentation,hypha and aerial hypha formation. The abundance of actinomycetes were 3,50 x104 – 71,50 x104CFU/g soil. Seven genus of actinomycetes were found, they were Streptomyces, Nocardia,Micromonospora, Microbiospora, Actinomadura, Actinoplanes, and Microtetraspora. Mostof them were distributed in 900 until 2500 m height above sea level. Streptomyces only knownin 2700, 2900 and 3057 m above sea level. Population of Streptomyces were predominant in allsoil types according to height, but were highest in 2500 m asl.Key words: Actinomycetes, fungi, Mount Ciremai, diversity, abundance.
Studi Kelimpahan Aktinomisetes Tanah dan Hubungannya Terhadap Enzim Selulase, Amilase, Total Karbon dan Nitrogen Hutan Pasca Kebakaran Bukit Bangkirai Kalimantan Timur Nurkanto, Arif
JURNAL BIOLOGI INDONESIA Vol 5, No 1 (2008): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (76.489 KB) | DOI: 10.14203/jbi.v5i1.3208

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ABSTRACTSoil Actinomycetes Population, Enzymes Activity, and its Relation with Carbon and NitrogenContent, in Bukit Bangkirai, East Kalimantan. Bukit Bangkirai is one of the tropical forest inIndonesia have been exposed with intense forest fire. The affected forest is subjectivelydivided into three level of damages, heavily damage forest (HD), low damage forest (LD) andcontrol (K). The objective of this research was to observe the abundance of Actinomyceteswhich have important role in ecological process. Through decompotition of organic materialsand nutriens cycle. Actinomycetes were isolated and enumerated by SDS-YE method. CFU/ gsoil (x 104) in K, HD and LD are 41,86 ± 25,52, 16,09 ± 5,70 and 18,96 ± 4,19 respectively. Amylaseand cellulase were determined by DNS method. Carbon and Nitrogen total were determinatedby CN analyzer. The different of amylase, cellulase activities and abundace of Actinomycetesbetween HD, LD, and Control plot were not significant. However, carbon and nitrogen totalare different. LD plot has the highest carbon and nitrogen total, followed by control and HDplot. There has no significant different among plot observed may indicate microbial communitiesof soil in Bukit Bangkirai have been recovered.Key words : actinomycetes, fire forest, amylase, cellulase, nitrogen total, carbon total
KERAGAMAN AKTINOMISETES KEPULAUAN WAIGEO, KABUPATEN RAJA AMPAT, PAPUA DAN POTENSINYA SEBAGAI PENDEGRADASI SELULOSADAN PELARUT FOSFAT Nurkanto, Arif
BERITA BIOLOGI Vol 9, No 1 (2008)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1170.905 KB) | DOI: 10.14203/beritabiologi.v9i1.776

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Some Actinomycetes isolated from Waigeo Raja Ampat Regency Papua have been identified. Those isolates were also characterized for their cellulolitic and phosphate solubilizing ability. Microscopic identification was based on Miyadoh (1997) and Holt (1994) methods.Actinomycetes could be identified by microscopic observation on spores, chain spore, hypha, aerial hypha and its pigmentation.The cellulolitic ability was observed by clear zone ratio in CMC medium and phosphate solubilizing activity by the same approach in Vikoskaya medium.From 139 Waigeos Actinomycetes isolates which had been deposited in LIPI Microbial Collection (LIPIMC) were identified as 10 genera (Actinomadura, Actinoplanes, Microbiospora, Micromonospora, Nocardia, Pseudonocardia, Saccharopolispora, Streptomices, Streptosporangium and Thermomonospora). As much as 57.5% phosphate solubilizing actinomycetes and 82.7 % cellulolitic actinomycetes were detected.
CELLULOLITIC ACTIVITIES OF ACTINOMYCETES ISOLATED FROM SOIL RHIZOSPERE OF WAIGEO, RAJA AMPAT, WEST PAPUA Nurkanto, Arif
Journal of Tropical Soils Vol 14, No 3: September 2009
Publisher : UNIVERSITY OF LAMPUNG

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.5400/jts.2009.v14i3.239-244

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Seven actinomycetes isolated from Waigeo?s soil rhizospere have been characterized in growth, cellulase activities and pH. These actinomycetes were chosen based on their capabilities in cellulose degradation which were recognized by previous study. Those isolates are belonging to Streptomyces and Actinoplanes genera based on morphological and molecular analyses. The growths of actinomycetes were investigated based on spectrophotometric approach and cellulase activities were conducted based on reduction sugar. Cellulase activities, cell growth, and pH had significant correlation based on statistical analysis.  The highest enzyme activities was acquired from  Streptomyces bobili LIPIMC-A-283 (0,519 µmol mL-1 minute-1) at 96 h of incubation. This study provided important basic information on potential cellulolitic alternative from actinomycetes group, as consideration on choosing low-cost technique using microbial agent in cellulose treatment.
KERAGAMAN AKTINOMISETES ASAL SERASAH, SEDIMEN, DAN TANAH PULAU ENGGANO, BENGKULU [Deversity of Actinomycetes From Soil, Sediment, and Leaf Litter Samples of Enggano Island, Bengkulu] Putri, Ade Lia; Nurkanto, Arif
BERITA BIOLOGI Vol 15, No 3 (2016)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (3223.38 KB) | DOI: 10.14203/beritabiologi.v15i3.2238

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A total of 344 isolates of actinomycetes were isolated from leaf litter and soil samples collected from four locations of Enggano Island, Bengkulu. eighty eight isolates were selected using morphological characteristics and identified using 16S rRNA gene sequence analysis. Three families (Micromonosporacea, Streptomycetaceae, and Streptosporangiaceae) and eight genera were found from these samples. Approximately 80% of these strains belonged to Streptomyces group and 20% of isolates were similar to rare actinomycetes. Based on homology search (BLAST) and phylogenetic tree analysis, isolates of rare actinomycetes (EgA15, EgA30, EgA85, EgA386, EgA236, EgA243, EgA335, EgA112, EgA41,EgA312, EgA314) were identified as Kitasatosporia paracochleata (100%), Kitasatospora azatica (99%), Kitasatospora griseola (99%), Sphaerisporangium album (99%), Actinoplanes nipponensis (99%), Pseudosporangium ferrugineum (99%), Nonomuraea rosela (99%), Nonomuraea guangzhouensis (98%), Micromonospora chaiyaphumensis (99%),Micromonospora chaiyaphumensis (99%), and Couchioplanes caeruleus (99%) respectively.
TOKSISITAS AKUT ORAL DUA SENYAWA BISANTRAKUINON (+)-2,2’-EPISITOSKIRIN A DAN (+)-1,1’-BISLUNATIN [ORAL ACUTE TOXICITY OF TWO BISANTHRAQUINONES (+)-2,2’-EPICYTOSKYRIN A AND (+)-1,1’-BISLUNATIN] Praptiwi, Praptiwi; Nurkanto, Arif; Wulansari, Dewi; Agusta, Andria
BERITA BIOLOGI Vol 14, No 1 (2015)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/beritabiologi.v14i1.1858

Abstract

Bisanthraquinones (+) - 2,2'-epicytoskyrin A and (+) -1,1'bislunatin produced by the endophytic fungus Diaporthe sp. GNBP-10 showed potent antibacterial activity on in-vitro test and have the opportunity to become new antibiotics candidates. The aspects of safety and toxicity of drug candidates have to be examined before applying to human. This study was conducted to determine the safety aspects of the compounds through acute oral toxicity testing in mice (Mus musculus). Acute toxicity of (+) - 2,2'-epicytoskyrin A and (+) - 1,1'-bislunatin evaluated by the method of Up and Down Procedure with limit test at a dose of 2000 mg / kg. Results of acute toxicity test showed that the LD50 of (+) - 2,2'-epicytoskyrin A and (+) - 1,1'-bislunatin were of 1638.87 mg / kg and > 2000 mg / kg respectively. Administration of (+)- 2,2'-epicytoskyrin A resulted in increased miliari multifocal hepatitis, fatty degeneration and necrosis of liver cells, and the renal tubule epithelial degeneration. Administration of (+) - 1,1'-bislunatin at a dose of 2000 mg / kg resulted in multifocal accumulation of inflammatory cells in the liver and degeneration of cells in the islets of Langerhans although not resulting in death. The administration of those compounds indicated the changes in the organs, but based on the UN/ECE classification of LD50 value showed that (+) 2,2'- epicytoskyrin A and (+) -1,1'-bislunatin included as low acute toxicity substance.
TOKSISITAS AKUT ORAL DUA SENYAWA BISANTRAKUINON (+)-2,2’-EPISITOSKIRIN A DAN (+)-1,1’-BISLUNATIN [ORAL ACUTE TOXICITY OF TWO BISANTHRAQUINONES (+)-2,2’-EPICYTOSKYRIN A AND (+)-1,1’-BISLUNATIN] Praptiwi, Praptiwi; Nurkanto, Arif; Wulansari, Dewi; Agusta, Andria
BERITA BIOLOGI Vol 14, No 1 (2015)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/beritabiologi.v14i1.1858

Abstract

Bisanthraquinones (+) - 2,2'-epicytoskyrin A and (+) -1,1'bislunatin produced by the endophytic fungus Diaporthe sp. GNBP-10 showed potent antibacterial activity on in-vitro test and have the opportunity to become new antibiotics candidates. The aspects of safety and toxicity of drug candidates have to be examined before applying to human. This study was conducted to determine the safety aspects of the compounds through acute oral toxicity testing in mice (Mus musculus). Acute toxicity of (+) - 2,2'-epicytoskyrin A and (+) - 1,1'-bislunatin evaluated by the method of Up and Down Procedure with limit test at a dose of 2000 mg / kg. Results of acute toxicity test showed that the LD50 of (+) - 2,2'-epicytoskyrin A and (+) - 1,1'-bislunatin were of 1638.87 mg / kg and > 2000 mg / kg respectively. Administration of (+)- 2,2'-epicytoskyrin A resulted in increased miliari multifocal hepatitis, fatty degeneration and necrosis of liver cells, and the renal tubule epithelial degeneration. Administration of (+) - 1,1'-bislunatin at a dose of 2000 mg / kg resulted in multifocal accumulation of inflammatory cells in the liver and degeneration of cells in the islets of Langerhans although not resulting in death. The administration of those compounds indicated the changes in the organs, but based on the UN/ECE classification of LD50 value showed that (+) 2,2'- epicytoskyrin A and (+) -1,1'-bislunatin included as low acute toxicity substance.