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Articles by issue : Vol 6, No 3 (2012): September 2012
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Articles
Phenolic Compounds of Sponge-associated Fungi (Lecanicillium evansii)

EFFENDI, HEFNI ( Centre for Environmental Research, Institut Pertanian Bogor, Indonesia )

Microbiology Indonesia Vol 6, No 3 (2012): September 2012
Publisher : Indonesian Society for microbiology

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Abstract

This study was chiefly aimed at pursuing new biologically active secondary metabolites of microfungus species, Lecanicillium evansii, isolated from sponge Callyspongia sp collected from West Bali Sea, Indonesia. Sponges were collected by scuba diving. A tiny piece of sponge was inoculated on the surface of malt agar plates and incubated at 27 ºC. In order to get a pure mono-culture of the fungus, repeated sub-culturing onto fresh malt agar plates were performed. The collected fungi were maintained on malt agar plates using the Wickerham medium. Mass cultivation of the fungus L. evansii (10 L) was carried out in 30 erlenmeyer flasks in Wickerham medium. After 10 days incubation, without shaking under constant room temperature (20 ºC), fungal mycelium were separated from the culture broth. The mycelia were extracted with methanol and ethyl acetate was added to the media. Both methanol-added mycelia and ethyl acetate-added media were left overnight. Seven compounds were isolated from L. evansii. Those compounds comprised phenolic compounds (terphenylin, deoxyterphenylin, terprenin 2, terprenin epoxide), bipeptide (cyclo-tyrosylprolyl), and simple aromatic compounds (acetyl hydroxybenzamide, 4-hydroxybenzaldehyde). Detailed analysis by NMR and mass spectrometry enabled their identification to be new deoxyterphenylin, new terprenin 2, and new terprenin epoxide.  

Methane and Nitrous Oxide Productions and Community Structure of Methanogenic Archaea in Paddy Soil of South Sulawesi, Indonesia

JUMADI, OSLAN ( Department of Biology, Faculty of Mathematics and Natural Science, Universitas Negeri Makassar , Jalan Daeng Tata Raya, Parangtambung, Makassar , South Sulawesi 90224, Indonesia ) , INUBUSHI, KAZUYUKI ( Graduate School of Horticulture, Chiba University , Matsudo, Chiba 271-8510, Japan )

Microbiology Indonesia Vol 6, No 3 (2012): September 2012
Publisher : Indonesian Society for microbiology

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Abstract

The potential of methane and nitrous oxide productions and community structure of methanogenic archaeahas been characterized by incubation studies and PCR-DGGE approaches, respectively. The results showed thataddition of straw into the soil was the most important factor that influenced CH4 production in this incubation 4 experiment, while N2O production was observed higher in soil without straw amendment under saturated 2 condition. Most of the clone sequences revealed from DGGE band was associated with a archaea lineage,Methanocellales ord. nov (Rice Cluster I). Other sequences belong to Methanomicrobiales, Methanosaetalesand Rice Cluster 2 (RC-2). The study also showed that the hydrogenotrophic methanogens were the dominant members in methanogenic archaea community obtained from tropical paddy field soil, while acetoclastic methanogens present in relatively minor quantity in the soil.

Molecular and Physiological Characterization of Copper-Resistant Bacteria Isolated from Activated Sludge in an Industrial Wastewater Treatment Plant in Rungkut-Surabaya, Indonesia

IRAWATI, WAHYU ( Department of Biology, Faculty of Science and Mathematics, Pelita Harapan University, Jalan M.H. Thamrin Boulevard 1100, Lippo Karawaci, Tangerang 15811, Banten ) , YUWONO, TRIWIBOWO ( Laboratory of Microbiology, Faculty of Agriculture, Gadjah Mada University, Yogyakarta ) , SOEDARSONO, JOEDORO ( Laboratory of Microbiology, Faculty of Agriculture, Gadjah Mada University, Yogyakarta ) , HARTIKO, HARI ( Laboratory of Biochemistry, Faculty of Biology, Gadjah Mada University, Yogyakarta )

Microbiology Indonesia Vol 6, No 3 (2012): September 2012
Publisher : Indonesian Society for microbiology

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Abstract

Copper resistant bacteria can be isolated from environments where copper levels are abundant from mining, industrial, or agricultural activities. The aim of this work was to study the molecular and physiologicalcharacteristics of indigenous copper resistant bacteria isolated from activated sludge in an industrial wastewatertreatment plant in Surabaya, Indonesia. The bacterial isolates were designated as strains IrC1, IrC2, and IrC4. Phylogenetic analysis based on 16S rDNA sequence analysis identified isolates IrC1, IrC2, and IrC4 as Acinetobacter oleivorans (98.41% similarity), Acinetobacter pitii (97.22% similarity), and Cupriavidus pauculus (96.99 similarity), respectively. The addition of 5 mM of CuSO4 in the medium affected morphological 4 appearance of all isolates to green and undulate margin might be due to the survival mechanism of bacteria by absorbing the copper. This studies indicated that copper resistance mechanism of all isolates was facilitated through the bioaccumulation of copper inside the cell, especially on the membrane fraction and inside the cytoplasm, albeit at a limited amount. It was observed that isolates IrC1, IrC2, and IrC4 were capable of accumulating 137.23 , 364.66 , and 272.07 mg L-1 of copper, respectively from the medium containing 8 mM CuSO4. The capability of isolates IrC1, IrC2, and IrC4 to accumulate copper can be exploited in bioremediation 4 process for removing copper from industrial sewage.

Decolorization of Orange II by Mixed Culture of Enterococcus faecalis ID6017 and Chryseobacterium indologenes ID6016

MEITINIARTI, VINCENTIA IRENE ( Faculty of Biology, Satya Wacana Christian University, Jalan Diponegoro 52-60, Salatiga 50712, Indonesia ) , TIMOTIUS, KRIS HERAWAN ( Krida Wacana Christian University, Jalan Arjuna Utara 6, Jakarta 11510, Indonesia ) , SOETARTO, ENDANG SUTARININGSIH ( Faculty of Biology, Universitas Gadjah Mada, Jalan Teknika Selatan, Yogyakarta 55281, Indonesia ) , SUGIHARTO, EKO ( Department of Chemistry, Universitas Gadjah Mada , Sekip Utara, Yogyakarta 55281, Indonesia )

Microbiology Indonesia Vol 6, No 3 (2012): September 2012
Publisher : Indonesian Society for microbiology

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Abstract

Previous work showed that Enterococcus faecalis ID6017 and Chryseobacterium indologenes ID6016 were able to decolorize the orange II qualitatively. In that experiment, E. faecalis could decolorize orange II more rapidly than C. indologenes. The objective of this study was to examine the decolorization of orange II by mixed culture and the growth of both bacterial species on orange II containing medium. The experiment was done in 500 mL sterilized Erlenmeyer flasks containing 285 mL growth media with 80 mg L-1 orange II. Five different treatments were performed in this project, i.e. medium was inoculated with (i) 15 mL of sterile aquadest, (ii) 15 mL of C. indologenes, (iii) 15 mL of E. faecalis, (iv) 7.5 mL of C. indologenes and 7.5 mL of E. faecalis, and (v) 7.5 mL of E. faecalis until decolorization occured , followed by inoculation with 7.5 mL of C. indologenes. Bacterial growth (total cells number), orange II, glucose, and suphanilic acid, as intermediate product of orange II decolorization, concentrations were measured every 2 h.The maximum decolorization of orange II was observed in the medium inoculated with a mixed culture of E. faecalis and C. indologenes. Decolorization of orange II occurred of growth and gave final concentration of sulphanilic acid of 7.06 mg L-1. During culture both species grow in equilibrium in terms of population.

Enzymatic Characterization of Recombinant Cyclodextrin Glycosyltransferase from Bacillus sp. a2-5a using Sagoo Starch as Substrate

IMANIAR, RINA ( School of Pharmacy, Institut Teknologi bandung, Jalan Ganesha 10, Bandung 40132, West Java, Indonesia ) , RIANI, CATUR ( School of Pharmacy, Institut Teknologi bandung, Jalan Ganesha 10, Bandung 40132, West Java, Indonesia ) , NATALIA, DESSY ( Faculty of Mathematics and Science, Institut Teknologi bandung, Jalan Ganesha 10, Bandung 40132, West Java, Indonesia ) , RETNONINGRUM, DEBBIE SOFFIE ( School of Pharmacy, Institut Teknologi bandung, Jalan Ganesha 10, Bandung 40132, West Java, Indonesia )

Microbiology Indonesia Vol 6, No 3 (2012): September 2012
Publisher : Indonesian Society for microbiology

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Abstract

Cyclodextrin (CD) is a cyclic oligosaccharide molecule and depending on the number of glucose molecules, three types of CDs are commonly used, α-CD, β-CD, and γ-CD. CDs can be produced enzymatically using starch as substrate catalyzed by CD glycosyltransferase (CGTase). In current research, recombinant CGTase production from the synthetic gene was optimized for its production using three growth media and two induction temperatures. The highest yield was obtained in Luria Bertani medium at 25 °C. The rCGTase protein was affinity purified as a 76.39 kDa protein which showed α-cyclization and starch hydrolysis activities using zymography method. The optimum temperature, pH and incubation time was 55 °C, 6, and 24 h, respectively. The enzyme was stable at a wide pHs in the range of 5-10, retained its half activity at 56 °C for 30 min and had cyclization ratio for α-CD : β-CD : γ-CD was 4 : 81 : 15. An amount of 542 mg β-cyclodextrin was produced from 100 mL reaction of 1% (b/v) sagoo starch using 38.4 μg rCGTase in optimum condition. This work reports for the first time the character of rCGTase from Bacillus sp. A2-5a using sagoo starch as a substrate.

Effects of Tetracycline and Temperature on Drosophila melanogaster Infected with Wolbachia Inducing the Popcorn-Effect

KUSMINTARSIH, ENDANG SRIMURNI ( Faculty of Biology, Universitas Jenderal Soedirman, Jalan Dr Soeparno 63 Grendeng, Purwokerto 53122, Indonesia )

Microbiology Indonesia Vol 6, No 3 (2012): September 2012
Publisher : Indonesian Society for microbiology

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Abstract

The expression of cytoplasmic incompatibility, parthenogenesis, and feminization in many hosts of Wolbachia is influenced by environmental factors such as temperature and antibiotics. Therefore, it might also affect Wolbachia inducing popcorn-effect. To examine the effects of temperature and antibiotic treatment on the life span of Drosophila melanogaster harbouring the popcorn-effect inducing strain of Wolbachia, flies were reared in different temperature such as at 20 and 29 °C, and cultured (from egg to adult stage) on a medium containing tetracycline. The tetracycline-treated Wolbachia was established by placing 0.25 mg m L-1 of tetracycline in the media. The result showed that there was no difference in the life span of D. melanogaster infected with Wolbachia popcorn-effect under untreated and treated condition with tetracycline at 20 °C. Therefore, there is no popcorn-effect in the D. melanogaster at low temperature (20 °C). While the life span of D. melanogaster at 29 °C, where infected flies have a shorter life span than treated flies. Popcorn-effect (shorter life span) was found at 29 °C.

Profile of Mutation on Drug Resistance Mycobacterium leprae Isolates in Indonesia Collected During 2003-2011

WAHYUNI, RATNA ( Leprosy Study Group, Institute of Tropical Disease, Universitas Airlangga, Jalan Mulyorejo Kampus C Unair, Surabaya 60115, Indonesia ) , ADRIATY, DINAR ( Leprosy Study Group, Institute of Tropical Disease, Universitas Airlangga, Jalan Mulyorejo Kampus C Unair, Surabaya 60115, Indonesia ) , ISWAHYUDI, ISWAHYUDI ( Leprosy Study Group, Institute of Tropical Disease, Universitas Airlangga, Jalan Mulyorejo Kampus C Unair, Surabaya 60115, Indonesia ) , PRAKOESWA, CITA ROSITA SIGIT ( Departement of Dermatology and Venereology , Faculty of Medicine, Universitas Airlangga ) , AGUSNI, INDROPO ( Departement of Dermatology and Venereology , Faculty of Medicine, Universitas Airlangga ) , IZUMI, SHINZO ( Leprosy Study Group, Institute of Tropical Disease, Universitas Airlangga, Jalan Mulyorejo Kampus C Unair, Surabaya 60115, Indonesia )

Microbiology Indonesia Vol 6, No 3 (2012): September 2012
Publisher : Indonesian Society for microbiology

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Abstract

Multidrug therapy (MDT) regiment has been used for leprosy all over the world for more than 20 years. Drug resistances of Mycobacterium have been reported from many areas. The resistance mostly occurred due to mutation on the gene coding protein targeted by anti-leprosy drugs. Two hundreds and seventy M. leprae isolates from some area in Indonesia were examined for studying the profile of mutation among isolates collected during 2003-2011. Drug resistance determining region of the folP1 gene and the rpoB gene was sequenced. The results showed 5 isolates of M. leprae harboured mutation only in the folP1 gene and another isolate harbored mutation in both the folP1 and rpoB gene. The point mutation in the folP1 gene that was found in 2 isolates occurred in codon 53 (ACC→GCC; Thr→Ala). Double point mutations on codon 53 that was found in two isolates were ACC→AGA (Thr→Arg) and ACC→AGG (Thr→Arg). The point mutation in the folP1 geneoccurred in codon 55 were found in two isolates were CCC→CTC (Pro→Leu) and CCC→CGC (Pro→Arg). Whereas mutation in the rpoB gene in one isolate occurred in codon 410 was GAT→TAT (Asp→Tyr). These mutations that altered the amino acids of the protein revealed that isolates of M. leprae were resistant to drug with variable profiles