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Rapid Assessment of Diverse Trichodermal Isolates of Indonesian Origin for Cellulase Production Fahrurrozi, Fahrurrozi; Ratnakomala, Shanti; Lisdiyanti, Puspita; Anindyawati, Trisanti; Sukara, Endang
Annales Bogorienses Vol. 14 No. 1 (2010): Annales Bogorienses
Publisher : BRIN

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Trichoderma is a well-known candidate to be promoted as cellulase producer for the hidrolysis of lignocellulose that contain in woody biomass. The number of trichodermal isolates in our laboratory collected from diverse ecosystem types in Indonesia increases significantly during the last 5 years. It is our aim to assess the cultures for its ability in producing cellulase. Sixty-six trichodermal isolates used in this experiment are obtained from Biotechnology Culture Collection (BTCC), Research Center for Biotechnology, Indonesian Institute of Sciences (LIPI). The 31 isolates were isolated from District of Liwa (South Sumatra, Indonesia) and the 35 isolates from District of Maros (South Sulawesi, Indonesia). The isolates were screened qualitatively, 7 isolates from Liwa and 12 isolates from Maros showed cellulolytic activity. From the results of quantitative test, two strains (ID08-T004 and ID08-T63) showed the higher cellulolytic activity among the selected strains, 133.5 and 133.5 U/ml, respectively. These extracellular enzymes were characterized their temperature and pH optimum. The temperature optimum for both enzymes was the same, 50C, with activity 213.6 U/ml for enzyme extracted from ID08-T004 and 197.3 U/ml for enzyme from ID08-T0063. The pH optimum was pH 5 of ID08-T004 with activity 137.7 U/ml and pH 6 for ID08-T063 with activity 75.0 U/ml. The enzymes from ID08-T004 and ID08-T063 were stable in their temperature and pH optimal condition even after 90 minutes incubation with activity 179.0 U/ml and 86.7 U/ml, respectively. The enzyme stability was approximately 150 minutes for both enzymes in the temperature and pH optimum.
Ecological Study of Rare-Actinomycetes in Soils and Leaf-Litters Lisdiyanti, Puspita; Ratnakomala, Shanti; Ridwan, Roni; Widyastuti, Yantyati; Otoguro, Misa; Ando, Katsuhiko
Annales Bogorienses Vol. 15 No. 2 (2011): Annales Bogorienses
Publisher : BRIN

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A total of 268 strains of Actinomycetes were isolated from soil and leaf-litter samples collected in Jambi (Sumatera) and Cibinong (West Java) using three selective isolation methods. All the isolates were identified by morphological characteristic and by analysis of 16S rDNA sequence. On the basis of their morphology and 16S rDNA sequence, 164 isolates were belonged to the Streptomyces Group and 104 isolates were belonged to the Rare-Actinomycetes (Non-Streptomyces) Group. Furthermore, 40% of isolates in Streptomyces Group and 62% of isolates in Non-Streptomyces Group are supposed to be new taxa. It is indicated the richness of Actinomycetes in these two area. The genus Streptomyces is the most abundant in soil samples, occupying 60-75% of all isolates; while the genus Actinoplanes is mainly found in leaf-litter samples (70%). The use of selective isolation media is important to elucidate the microbial diversity.
Diversity of Actinomycetes from Soil Samples Collected from Lombok Island, Indonesia Lisdiyanti, Puspita; Tamura, Tomohiko; Ratnakomala, Shanti; Ridwan, Roni; Kartina, Gina; Lestari, Yulin; Ando, Katsuhiko; Widyastuti, Yantyati
Annales Bogorienses Vol. 16 No. 1 (2012): Annales Bogorienses
Publisher : BRIN

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A total of 137 strains of Actinomycetes were isolated from 14 soil samples collected in 3 part of Lombok Island, Indonesia using SDS-YE (SY) and Rehydration and Centrifugation (RC) isolation methods and Humic Acid Vitamin (HV) agar as isolation media. All the isolates were identified by morphological characteristic and by analysis of 16S rRNA gene sequence. On the basis of their morphology and 16S rRNA gene sequence, 67% of isolates were belonged to the Streptomyces Group and 33% of isolates were belonged to the Rare-Actinomycetes (Non-Streptomyces) Group. Nine families and 15 genera were found from the samples. It is indicated the richness of actinomycetes in these area. The genus Streptomyces is the most abundant in all soil samples, occupying 67% of all isolates by using SY method; while the genus Actinoplanes is mainly found by using RC as isolation method. Two strains belong to the genus Virgisporangium and Catenulispora that showed taxonomic interest need further study for describing as ne species.
Optimization of Culture Conditions for Production of β-Mannanase by Strain Nonomuraea sp. ID06-379 using Submerged Substrate Fermentation Ratnakomala, Shanti; Yopi, Yopi; Suhartono, Maggy Thenawidjaja; Meryandini, Anja; Prasetya, Bambang
Annales Bogorienses Vol. 18 No. 2 (2014): Annales Bogorienses
Publisher : BRIN

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The objective of this study was to investigate the effect of media compositions on the production of β-mannanase by Nonomuraea sp. ID06-379. The study was focused on the influence of carbon, nitrogen, phosphorus and detergents on β-mannanase synthesis through manipulating media compositions on production medium. The results indicated that for carbon sources, locus bean gum (0.745 ± 0.036 U/ml) showed maximum mannanase activity. Malt extract was the best nitrogen source for producing β-mannanase (1.075 ± 0.006 U/ml), (NH4)2HPO4 as phosphate source (1.733 ± 0.026 U/ml) and Tween 80 (1.145 ± 0.003 U/ml) as surfactants effect on increasing permeability of bacterial cell membrane, enhancing membrane transport and excretion of extracellular enzymes into the production media. The results showed that 1% malt extract, 0.5% locus bean gum and 0.05% (NH4)2HPO4 were good substances for nitrogen source, carbon source and phosphate respectively. The highest production of β-mannanase by Nonomuraea sp. ID06-379 (5.33 U/mg) was reached in the medium optimization (Vogel’s minimal medium) contained the following ingredients: 0.5% locus bean gum, 1% malt extract and 0.05% (NH4)2HPO4, under submerged fermentation with shaking at 120 rpm and 28C for 2 days incubation.