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POTENCY OF BIOCONTROL AGENTS ISOLATED FROM COMPOST AND PEAT SOIL OF TROPICAL PEAT SWAMP FOREST IN KALAMPANGAN ZONE, CENTRAL KALIMANTAN Yuliar, Yuliar; Abidin, Zaenal; Mangunwardoyo, Wibowo
Indonesian Journal of Forestry Research Vol 8, No 2 (2011): Journal of Forestry Research
Publisher : Secretariat of Forestry Research and Development Agency

Show Abstract | Download Original | Original Source | Check in Google Scholar

Rhizoctonia solani is a soil pathogen that causes diseases in wide range of hosts of agricultural, horticultural and flower crops. Biological control is the most promising way for the diseases management and it is environment friendly too. The objective of this study was to isolate and screen the potency of soil bacteria as biological control from various local compost and peat soil of tropical peat swamp forest in Kalampangan Zone, Central Kalimantan. Forty seven isolates from peat soil and compost were screened for biocontrol agent of Rhizoctonia solani . R. Solani Seven out of thirteen peat soil isolates, and six out of thirty three compost isolates showed antagonistic activity against in Potato Dextrose Agar. The cultivation of the antagonistic isolates in Trypticase Soy Broth (TSB) was extracted and analysed by high performance liquid chromatography (HPLC) column. The HPLC analyzes indicated that the antagonistic isolates produce an antifungal iturin A. Macroscopic observation of isolates colonies showed that form of their colonies were amuboid, myceloid, curled, circular, rhizoid, irregular and filamentous. These achievement indicate peat swamp forest not only offer a potential biocontrol agents of damping off but also provide a new source for production of antibiotics.

Variasi Intraspesies Lactobacillus plantarum (Orla-Jensen) Bergey et al. Asal Sayur Asin Berdasarkan Analisis Molekuler ., Sulistiani; ., Abinawanto; Sukara, Endang; Dinoto, Achmad; Mangunwardoyo, Wibowo
JURNAL BIOLOGI INDONESIA Vol 11, No 1 (2015): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v11i1.2162

The current study is the first report on intraspecies analysis of L. plantarum from sayur asin in Indonesia using molecular approach. Three molecular techniques, i.e., restriction fragment length polymorphism (RFLP) 16S-23S rDNA intergenic spacer region (ISR), random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) and enterobacterial repetitive intergenic consensus (ERIC-PCR) were used to determine the intraspecies diversity of L. plantarum responsible for spontaneous fermentation in sayur asin. These methods were aimed to discriminate 46 isolates of L. plantarum isolated from sayur asin, including the type strain. PCR amplification of the 16S-23S rDNA ISR revealed two-bands profile of 800 and 600 bp specific to lactobacilli. RAPD-PCR and ERIC-PCR were very valuable in discriminating genetic polymorphism among L. plantarum isolates by producing bands ranged from 4-10 bands (360-2620 bp) and 6-12 bands (160-2900 bp), respectively. Dendograms generated from UPGMA cluster analysis based on RAPD-PCR and ERIC-PCR data showed that all isolates were grouped into three major clusters with 74% and 68.6% genetic similarity thresholds, respectively.The study indicated that strains belong to L. plantarum isolated from sayur asin were divided into three genotypic groups.ÃÂ Keywords: ERIC-PCR, Intraspecies, Lactobacillus plantarum, RAPD-PCR, RFLP 16S-23S rDNA ISRÃÂ

OPTIMASI DAN PEMEKATAN LIPASE Bacillus halodurans CM1 Aisyah, Arina; Mangunwardoyo, Wibowo; Trismilah, Trismilah; Suhendar, Dadang
Al-Kauniyah: Jurnal Biologi Vol 10, No 2 (2017): Al-Kauniyah Jurnal Biologi
Publisher : Department of Biology, Faculty of Science and Technology, Syarif Hidayatullah State Islami

Abstrak Lipase diketahui memiliki peranan penting dalam bidang industri. Produksi lipase dapat dihasilkan oleh kapang, khamir, dan bakteri. Penelitian bertujuan untuk meningkatkan aktivitas lipase yang dihasilkan oleh Bacillus halodurans CM1. Aktivitas lipase dapat ditingkatkan dengan optimasi komposisi media, mutasi bakteri dengan radiasi gamma dan N-methyl-Nâ€™-nitro-N-nitrosoguanidine (NTG). Enzim yang dihasilkan dipekatkan dengan metode stirred-cell ultrafiltration (UF)-ammonium sulfat dan UF-Polyethylene glycol (PEG). Uji aktivitas dilakukan pada tujuh media yang berbeda untuk mendapatkan media produksi. Delapan variabel komposisi media dioptimasi dengan rancangan Plackett-Burman. Bakteri dimutasi dengan radiasi gamma dosis 0,1â€“0,4 kGy dan NTG 0,05â€“0,15 mg/mL dengan waktu inkubasi 1â€“3 jam. Hasil penelitian menunjukkan bahwa media produksi yang digunakan berdasarkan optimasi media dan komposisi media Plackett-Burman adalah media dasar Bora & Bora yang mengandung 0,5% palm oil (PO) dan 0,09% CaCl2. Aktivitas lipase optimal diproduksi oleh bakteri hasil mutasi dengan NTG 0,1 mg/mL yang diinkubasi selama 3 jam. Pemekatan enzim UF-ammonium sulfat dan UF-PEG mampu meningkatkan aktivitas enzim lipase sebesar 18,44%.Â Â Abstract Lipase is known to have an important role in the industrial field. Lipase can be produced by molds, yeasts, and bacteria. The research aimed to increase the activity of lipase produced by Bacillus halodurans CM1. Lipase activity can be improved by optimization of the composition of the media, the mutation of bacteria with gamma radiation and N-methyl-N'-nitro-N-nitrosoguanidine (NTG). The enzyme was concentrated by stirred-cell ultrafiltration method (UF)-ammonium sulfate and UF-Polyethylene glycol (PEG). The activity test was performed on seven different media to get production media. The eight variables of the media composition were optimized by Plackett-Burman design. The bacteria were subject to mutation by using 0.1â€“0.4 kGy dose of gamma radiation and 0.05â€“0.15 mg/mL NTG with incubation time for 1â€“3 hours. The results showed that the production media used based on optimization and composition of Plackett-Burman media was Bora Bora medium that containing 0.5% palm oil (PO) and 0.09% CaCl2. Optimum lipase activity was produced by the bacterium that mutated with 0.1 mg/mL NTG, incubated for 3 hours. The concentrated by UF-ammonium sulfate and UF-PEG could increase the lipase activity by 18.44%.

POTENCY OF BIOCONTROL AGENTS ISOLATED FROM COMPOST AND PEAT SOIL OF TROPICAL PEAT SWAMP FOREST IN KALAMPANGAN ZONE, CENTRAL KALIMANTAN Yuliar, Yuliar; Abidin, Zaenal; Mangunwardoyo, Wibowo
Indonesian Journal of Forestry Research Vol 8, No 2 (2011): Journal of Forestry Research
Publisher : Secretariat of Forestry Research and Development Agency

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20886/ijfr.2011.8.2.144-157

Rhizoctonia solani is a soil pathogen that causes diseases in wide range of hosts of agricultural, horticultural and flower crops. Biological control is the most promising way for the diseases management and it is environment friendly too. The objective of this study was to isolate and screen the potency of soil bacteria as biological control from various local compost and peat soil of tropical peat swamp forest in Kalampangan Zone, Central Kalimantan. Forty seven isolates from peat soil and compost were screened for biocontrol agent of Rhizoctonia solani . R. Solani Seven out of thirteen peat soil isolates, and six out of thirty three compost isolates showed antagonistic activity against in Potato Dextrose Agar. The cultivation of the antagonistic isolates in Trypticase Soy Broth (TSB) was extracted and analysed by high performance liquid chromatography (HPLC) column. The HPLC analyzes indicated that the antagonistic isolates produce an antifungal iturin A. Macroscopic observation of isolates colonies showed that form of their colonies were amuboid, myceloid, curled, circular, rhizoid, irregular and filamentous. These achievement indicate peat swamp forest not only offer a potential biocontrol agents of damping off but also provide a new source for production of antibiotics.

ANALISIS SENYAWA BIO AKTIF DARI EKSTRAK BUI PICUNG (Pangium edule Reinw.) SEGAR Mangunwardoyo, Wibowo; Ismaini, Lily; Heruwati, Endang Sri
BERITA BIOLOGI Vol 9, No 3 (2008)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Pangi (Pangium edule Reinw.) seed has long been used traditionally as a preservative agent for fish, especially in remote areas.A study has been conducted on analysis of bioactive compounds of pangi fresh seeds extracts and their fractions.In this study,maceration of fresh seed using water and 50% ethanol was carried out followed by thin layer chromatography (TLC) analysis to see weather the extracts contained tannin. Both extract then separated into their respective fractions using column chromatography.Fractions which had been tested to have highest antibacterial activity were then analysed using gas chromatography mass spectrometry (GC-MS) to assess the active compounds which believe to be a preservative agent.Identification of water and 50% ethanol extract of Pangium edule Reinw.fresh seeds with TLC resulted that tannin were found in those extracts. Furthermore, GC-MS analysis showed that fractions which had been previously tested to have high antibacterial activity contained 9-octadecanoic acid whith similarity index of 89%, and 1,2-benzenedicarboxylic acid whith similarity index of 94-85%.

VARIASI GENETIK Lactobacillus fermentum Beijerink ASAL SAYUR ASIN BERDASARKAN ANALISIS RFLP 16S-23S rDNA ISR, RAPD-PCR DAN ERIC-PCR [Genetic Variation of Lactobacillus fermentum Beijerink Origin Sayur Asin Based on RFLP 16S-23S rDNA ISR, RAPD-PCR and ERIC-PCR Analysis] Sulistiani, Sulistiani; Mangunwardoyo, Wibowo; Abinawanto, Abinawanto; Sukara, Endang; Dinoto, Achmad; Salamah, Andi
BERITA BIOLOGI Vol 16, No 2 (2017)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Molecular analysis of Lactobacillus fermentum isolates is essential to understand their genetic variation in relations to their roles in sayur asin fermentation process. Combination of three molecular techniques which is restriction fragment length polymorphism (RFLP) of 16S23S rDNA intergenic spacer region (ISR), random amplified polymorphic DNA (RAPD-PCR) and an enterobacterial repetitive intergenic consensus (ERIC-PCR) analysis were performed to discriminate 19 representative isolates of L. fermentum isolated from sayur asin. The result showed that L. fermentum strain D11 is distantly related to other isolates based on RFLP using HhaI restriction enzyme and RAPDPCR analyses. In addition, both of RAPD-PCR and ERIC-PCR successfully determined the genetic variation among L. fermentum strains by exhibiting distinct 4-8 bands (800-2080 bp) and 4-10 bands (280-3050 bp), respectively. A dendogram generated from UPGMA cluster analysis of both RAPD-PCR and ERIC-PCR data showed two distinct genotypic groups exist among L. fermentum isolated from sayur asin in Indonesia.

Enhancement of Î²-Glucosidase Activity in Penicillium sp. by Random Mutation with Ultraviolet and Ethyl Methyl Sulfonate Syafriana, Vilya; Nuswantara, Sukma; Mangunwardoyo, Wibowo; Lisdiyanti, Puspita
ANNALES BOGORIENSES Vol 18, No 2 (2014): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

The genus Penicillium has a potential ability to produce Î²-glucosidase. The aim of the study was to improve the Î²-glucosidase activity of Penicillium sp. ID10-T065 with physical (Ultraviolet = UV), chemical (Ethyl Methyl Sulfonate = EMS), and combined mutation (UV-EMS). The spores of Penicillium sp. ID10-T065 were exposed into UV irradiation for 3 minutes with dose of 0.1 J/cm2 and 13 cm of distances. Chemical mutation was done by treated spores into 3% of EMS solution for an hour. Combined mutation of UV and EMS were also performed by UV for 3 minutes (0.1 J/cm2, 15 cm) and continued with soaking into 2-3% of EMS solution. The developed mutants were screened, selected and assayed. Comparison of enzyme activities with the wild- type (1.78 U/ml), mutant UV13 (5.53 U/ml) showed a 3.1 fold increase; mutant EM31 (4.26 U/ml) showed a 2.4 fold increase. Meanwhile, mutant UM23 obtained from the multiple exposures showed a decreased activity (1.75 U/ml). Mutant UV13 showed the best enzyme activity to be considered as a potential strain for Î²-glucosidase producer. This result needs to be further elaborated especially on its genetic stability studies in order for the ascertained as a stable mutant.

IDENTIFICATION AND CULTIVATION OF MFW 23-08 ISOLATED FROM MARINE SPONGES FOR BIOACTIVE COMPOUND PRODUCTION Chasanah, Ekowati; Pratitis, Asri; Mangunwardoyo, Wibowo
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 7, No 2 (2012): August 2012
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v7i2.16

Production of marine bioactive compound for commercial usage has been hampered due tothe problem of raw material supply. To overcome this, marine microbes especially those associatedwith the bioactive-compound producer biota, has been explored as bioactive sources, with severaladvantages such as shorter production time, cheaper production cost and avoiding over exploitationof marine biota sources. Previous research showed that fungi MFW 23-08 was one of the potentialisolates from Wakatobi sponges which produced bioactive compounds that was active againstbreast cancer cell line and as antioxidant. This study was intended to identify MFW 23-08 andoptimize the production of its bioactive compound through optimization of MFW 23-08 culture.Culture optimization was conducted using 3 liquid media, i.e. malt extract broth (MEB), glucosepeptone yeast (GPY), and minimal fungal media (MFM), and cultivation periods, i.e. 2, 4, 6, 8, and10 wk. Results revealed that MFW 23-08 crude extract of 2 wk-MFM cultivation, at the concentrationof 30 Î¼g/ml, was able to retard 87% breast cancer (T47D) cell growth. While at concentration of100 Î¼g/ml, the 6 wk. MEB cultivated extract was able to hamper free radicals (56%). However, thecrude extract from MFM media cultivation, in the concentration of 50 and 100 Î¼g/ml was not able toinhibit Escherichia coli and Staphylococcus aureus growth. Based on molecular identificationusing ITS1-ITS4 primers, MFW 23-08 isolate was 99% similar to Â Penicillium citrinum, P.griseofolvum and Penicillium Â sp.

Improvement of Endoglucanase Activity in Penicillium oxalicum ID10-T065 Mutated by Ultra Violet Irradiation and Ethidium Bromide Caniago, Asnany; Mangunwardoyo, Wibowo; Nuswantara, Sukma; Lisdiyanti, Puspita
ANNALES BOGORIENSES Vol 19, No 2 (2015): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Penicillium sp. is known as filamentous fungi that produce complete cellulase. Cellulase. This study aims to improve endoglucanase activity of Penicillium oxalicum ID010-T065 by mutated with ultra violet irradiation (with dose of 0.1 J/cm2, 15 cm), ethidium bromide (10 Âµg/mL, 1 hour) and combination of both mutagens. The endoglucanase activity of all mutants was higher than that of the wild type (1.03 U/mL). Mutant UVEB-42 exposed to combine mutation showed the highest endoglucanase activity (2.76 U/mL) with a 2.70 fold increase. Mutant EB-45 (1.83 U/mL) exposed to ethidium bromide solution showed a 1.8 fold increase. Mutant UV-13 (1.72 U/mL) exposed to UV irradiation for 3 minutes showed a 1.7 fold increase. All mutants have optimum endoglucanase activity at 50 Â°C. Mutant UVEB-53 showed the highest thermostability by retaining 86 % of endoglucanase activity at 90 Â°C. The gene analysis of the endoglucanase I gene showed 3 bases mutated at mutant UV-13 and UVEB-53 that changed proline to serine. Mutant EB-45 showed 4 bases mutated that changed valine to glysine and proline to serine. Two bases mutated at Mutant UVEB-53 changed proline to serine. Bases mutated in eg1 gene could influenced the enhance of enzym activity in mutant.

IDENTIFIKASI MOLEKULAR ISOLAT KAPANG PENGHASIL Ã-GLUCAN BERDASARKAN DAERAH INTERNAL TRANSCRIBED SPACER (ITS) Srikandace, Yoice; Caterina A, Ines Irene; Mangunwardoyo, Wibowo
BERITA BIOLOGI Vol 9, No 5 (2009)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Research was conducted to identify the fungal isolate G.23 which produces b-glucan from the Biopharmacy Laboratoriums collection.Indonesian Institute of Sciences (LIP1) based on the sequence of the Internal Transcribed Spacer (ITS) regions.DNA was isolated from mycelia and the ITS region was amplified by Polymerase Chain Reaction (PCR) with ITS1 and ITS4 primers. The PCR product was purified using the QIAquick PCR Purification kit (Qiagen). BigDye terminator cycle sequencing Ready Reaction Kit (Perkin Elmer Applied Biosystem) was used and the product was purified with the AutoSEQ G-50 Kit (Qiagen).The sequence obtained analysed using Basic Local Alignment Search Tool nucleotide (BLAST)n homology search.The BLASTn result showed that the fungal isolate G.23 belongs to the genus Aspergillus. Taxa closely related to this isolate were Aspergillus elegans,A. ochraceus and A.sclerotiorum with 96% sequence homology.ClustalX was used for sequence-alignment. Phylogenetic analysis was constructed using the Neighbour Joining (NJ) method with Kimura two parameters. The phylogenetic tree obtained showed that fungal isolate G.23 separated from A. elegans, A. ochraceus and A. sclerotiorum which indicated that fungal G.23 belonged to a different spesies.Morphological observation on culture and microscopic appearance of the fungal isolate G.23 supported that this isolate differs from A. elegans, A. ochraceus and A. sclerotiorum.

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