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Winda Ayu Syafitri
Abstrak :
Tujuan penelitian adalah mengetahui aktivitas amilolitik 17 isolat 'Actinobacteria' termofilik pada suhu tinggi, dan memperoleh informasi spesies, dan posisi filogenetik isolat potensial berdasarkan data sekuens gen 16S rRNA, analisis filogenetik, karakterisasi morfologi, fisiologi, dan biokimia. Kemampuan tumbuh 17 isolat 'Actinobacteria' termofilik pada berbagai variasi suhu diuji menggunakan medium ISP 1 agar, dan diinkubasi pada suhu 45, 50, 55, 60 oC selama 7 hari. Berdasarkan hasil penelitian, 17 isolat memiliki pertumbuhan yang bervariasi pada suhu 45--60 oC. Tujuh belas isolat tumbuh pada suhu inkubasi 45 oC, 16 isolat pada suhu 50 oC, enam isolat pada suhu 55 oC, dan lima isolat pada suhu 60 oC terdiri atas SL1-2-R-2, SL1-2-R-3, SL1-2-R-4, SL2-2-R-15, dan SL3-1-R-16. Aktivitas amilolitik 17 isolat 'Actinobacteria' termofilik pada berbagai variasi suhu diuji dengan metode 'starch agar plate', menggunakan medium Minimal (Mm) agar dengan penambahan pati ('soluble starch') sebagai substrat sebanyak 1% (b/v), dan diinkubasi pada suhu 45, 50, 55, dan 60 oC selama 7 hari. Aktivitas amilolitik yang positif ditandai dengan terbentuknya zona bening di sekitar koloni bakteri setelah diteteskan larutan 'Lugol's iodine' sebanyak 1,5 ml. Hasil penelitian menunjukkan bahwa sebagian besar isolat yang diperoleh dari tanah di dekat geiser Cisolok memiliki aktivitas amilolitik yang bervariasi pada suhu 45--60 oC. Lima belas isolat memiliki aktivitas amilolitik pada suhu 45 oC, 13 isolat pada suhu 50 oC, empat isolat pada suhu 55 oC, dan hanya tiga isolat pada suhu 60 oC. Namun demikian, dua isolat (SL2-2-R-15 dan SL3-1-R-16) tidak memiliki aktivitas amilolitik pada suhu 45, 50, dan 55 oC setelah diinkubasi selama 7 hari. Tiga isolat potensial yang memiliki aktivitas amilolitik pada suhu 60 oC (SL1-2-R-2, SL1-2-R-3, dan SL1-2-R-4), berdasarkan data sekuens gen 16S rRNA, analisis filogenetik, dan karakterisasi fenotipik tiga isolat potensial tersebut diidentifikasi sebagai 'Actinomadura keratinilytica'. ...... The aims of this study were to screen for amylolytic activity of the 17 themorphilic 'Actinobacteria' at high temperature, and to obtain species information and phylogenetic position based on 16S rRNA gene sequence, phylogenetic analysis, morphological, physiological, and biochemical characterizations. The ability to grow at various temperature was carried out on ISP 1 agar medium, incubated at 45, 50, 55, 60 oC for 7 days. The results showed that the 17 isolates 'Actinobacteria' have varying growth at a temperature of 45--60 oC. Seventeen, 16, and six isolates grew at 45, 50, and 55 oC, respectively, and only five isolates grew at 60 oC, designated SL1-2-R-2, SL1-2-R-3, SL1-2-R-4, SL2-2-R-15, and SL3-1-R-16. Amylolytic activity of the 17 themorphilic 'Actinobacteria' at various temperature was carried out using the starch agar plate method on Minimal (Mm) agar medium with the addition of 1% (w/v) soluble starch as substrate, and incubated at 45, 50, 55, and 60 oC for up to 7 days. Amylolytic activity was detected by flooding the plates with 1.5 ml of Lugol's iodine solution. Clear zones around the colonies indicated positive results for amylolytic activity. The results showed that most of the isolates obtained from the soil near the Cisolok geyser have varying amylolytic activity at a temperature of 45--60° C. In this study, 15, 13, four, and three out of 17 isolates were positive for amylolytic activity at 45, 50, 55, and 60 oC, respectively. Meanwhile, two isolates, designated SL2-2-R-15 and SL3-1-R-16, showed negative results for amylolytic activity at 45, 50, and 55 oC, even after 7 days of incubation. Three potential isolates which have amylolytic activity at 60 oC (designated SL1-2-R-2, SL1-2-R-3, and SL1-2-R-4), based on 16S rRNA gene sequence, phylogenetic analysis, and phenotypic characterizations were identified as 'Actinomadura keratinilytica.'
Depok: Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Indonesia, 2019
T53675
UI - Tesis Membership  Universitas Indonesia Library
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Fitrianingsih
Abstrak :
The aims of this study are to provide data regarding the taxonomic study of thermophilic Actinobacteria based on 16S rRNA gene sequences, description, and assessment for secondary metabolite biosynthetic gene clusters (BGCs) in the genome of novel taxa, and its antibacterial activity. Thirty-one isolates of thermophilic Actinobacteria were isolated from soil samples in Cisolok geothermal area, West Java. The 16S rRNA gene sequence-similarity search against all related species was performed using EzTaxon-e database. The sequences of 31 isolates showed similarity to member of family Micromonosporaceae, Nocardiaceae, Pseudonocardiaceae, Streptomycetaceae, Streptosporangiaceae, and Thermomonosporaceae. Six isolates displayed high similarity to genera in the family Pseudonocardiaceae, and most closely related to the genus Thermotunica, T. guangxiensis AG2-7T with similarity values from 94.6 to 95.2%. Phenotypic features and phylogenetic data differentiated strain SL3-2-4T from members of the family Pseudonocardiaceae. Therefore, the strain SL3-2-4T is proposed as a representative of a novel species in a novel genus, Gandjariella thermophila gen. nov., sp. nov. The genome of SL3-2-4T contained 21 antiSMASH-identified secondary metabolite regions harboring BGCs. These BGCs were for polyketide synthase, non-ribosomal peptide synthase, and ribosomally synthesized and post-translationally modified peptide family clusters. Thirteen and five regions displayed low (4–35%) and no similarity with known BGCs for secondary metabolites, respectively. Screening for antibacterial activity showed that strains SL3-2-4T and SL3-2-7 on MM 2 medium solidified with gellan gum at 45°C for 14 days demonstrated inhibitory activity against all Gram-positive, but not Gram-negative bacteria. Strain SL3-2-10 on ISP 3 gellan gum medium incubated for seven-days only active against K. rhizophila NBRC 12078. The results indicated that novel taxa have the potential for the discovery of active secondary metabolites.
Depok: Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Indonesia, 2022
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UI - Disertasi Membership  Universitas Indonesia Library
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Winda Ayu Syafitri
Abstrak :
Tujuan penelitian adalah mengetahui aktivitas amilolitik 17 isolat Actinobacteria termofilik pada suhu tinggi, dan memperoleh informasi spesies, dan posisi filogenetik isolat potensial berdasarkan data sekuens gen 16S rRNA, analisis filogenetik, karakterisasi morfologi, fisiologi, dan biokimia. Kemampuan tumbuh 17 isolat Actinobacteria termofilik pada berbagai variasi suhu diuji menggunakan medium ISP 1 agar, dan diinkubasi pada suhu 45, 50, 55, 60 °C selama 7 hari. Berdasarkan hasil penelitian, 17 isolat memiliki pertumbuhan yang bervariasi pada suhu 45-60 °C. Tujuh belas isolat tumbuh pada suhu inkubasi 45 °C, 16 isolat pada suhu 50 °C, enam isolat pada suhu 55 °C, dan lima isolat pada suhu 60 °C terdiri atas SL1-2-R-2, SL1-2-R-3, SL1-2-R-4, SL2-2-R-15, dan SL3-1-R-16. Aktivitas amilolitik 17 isolat Actinobacteria termofilik pada berbagai variasi suhu diuji dengan metode starch agar plate, menggunakan medium Minimal (Mm) agar dengan penambahan pati (soluble starch) sebagai substrat sebanyak 1% (b/v), dan diinkubasi pada suhu 45, 50, 55, dan 60 °C selama 7 hari. Aktivitas amilolitik yang positif ditandai dengan terbentuknya zona bening di sekitar koloni bakteri setelah diteteskan larutan Lugol's iodine sebanyak 1,5 ml. Hasil penelitian menunjukkan bahwa sebagian besar isolat yang diperoleh dari tanah di dekat geiser Cisolok memiliki aktivitas amilolitik yang bervariasi pada suhu 45-60 °C. Lima belas isolat memiliki aktivitas amilolitik pada suhu 45 °C, 13 isolat pada suhu 50 °C, empat isolat pada suhu 55 °C, dan hanya tiga isolat pada suhu 60 °C. Namun demikian, dua isolat (SL2-2-R-15 dan SL3-1-R-16) tidak memiliki aktivitas amilolitik pada suhu 45, 50, dan 55 °C setelah diinkubasi selama 7 hari. Tiga isolat potensial yang memiliki aktivitas amilolitik pada suhu 60 °C (SL1-2-R-2, SL1-2-R-3, dan SL1-2-R-4), berdasarkan data sekuens gen 16S rRNA, analisis filogenetik, dan karakterisasi fenotipik tiga isolat potensial tersebut diidentifikasi sebagai Actinomadura keratinilytica. ......The aims of this study were to screen for amylolytic activity of the 17 themorphilic Actinobacteria at high temperature, and to obtain species information and phylogenetic position based on 16S rRNA gene sequence, phylogenetic analysis, morphological, physiological, and biochemical characterizations. The ability to grow at various temperature was carried out on ISP 1 agar medium, incubated at 45, 50, 55, 60 °C for 7 days. The results showed that the 17 isolates Actinobacteria have varying growth at a temperature of 45-60 °C. Seventeen, 16, and six isolates grew at 45, 50, and 55 °C, respectively, and only five isolates grew at 60 °C, designated SL1-2-R-2, SL1-2-R-3, SL1-2-R-4, SL2-2-R-15, and SL3-1-R-16. Amylolytic activity of the 17 themorphilic Actinobacteria at various temperature was carried out using the starch agar plate method on Minimal (Mm) agar medium with the addition of 1% (w/v) soluble starch as substrate, and incubated at 45, 50, 55, and 60 °C for up to 7 days. Amylolytic activity was detected by flooding the plates with 1.5 ml of Lugol's iodine solution. Clear zones around the colonies indicated positive results for amylolytic activity. The results showed that most of the isolates obtained from the soil near the Cisolok geyser have varying amylolytic activity at a temperature of 45-60 °C. In this study, 15, 13, four, and three out of 17 isolates were positive for amylolytic activity at 45, 50, 55, and 60 °C, respectively. Meanwhile, two isolates, designated SL2-2-R-15 and SL3-1-R-16, showed negative results for amylolytic activity at 45, 50, and 55 °C, even after 7 days of incubation. Three potential isolates which have amylolytic activity at 60 °C (designated SL1-2-R-2, SL1-2-R-3, and SL1-2-R-4), based on 16S rRNA gene sequence, phylogenetic analysis, and phenotypic characterizations were identified as Actinomadura keratinilytica.
Depok: Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Indonesia, 2019
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UI - Tesis Membership  Universitas Indonesia Library
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Putri Pratiwi Setyaningsih
Abstrak :
Tujuan dari penelitian ini adalah untuk memperoleh isolat 'Actinobacteria' termofilik dari tanah di sekitar geiser Cisolok, Jawa Barat yang memiliki aktivitas selulolitik pada suhu tinggi serta mengetahui posisi filogenetik isolat terpilih terhadap spesies-spesies terdekatnya berdasarkan gen 16S rRNA. Penapisan kemampuan degradasi selulosa 17 isolat dilakukan secara kualitatif pada 'Minimal medium' (Mm) padat yang ditambahkan substrat yaitu 'carboxymethyl cellulose' (CMC) 1% (b/v) atau  'microcrystalline cellulose' (MCC) 1% (b/v) kemudian diinkubasi selama 7 hari. Pengamatan dilakukan dengan pewarnaan 'Congo red' 0,2% (b/v) dan zona bening pada sekitar koloni mengindikasikan degradasi substrat. Hasil penapisan menunjukkan bahwa 15 isolat mendegradasi CMC 1% dan 12 isolat mendegradasi MCC 1% pada suhu 45 oC, 14 isolat mendegradasi CMC 1% dan MCC 1% pada suhu 50 oC, 4 isolat mendegradasi CMC 1% dan MCC 1% pada suhu 55 oC, dan 3 isolat mendegradasi CMC 1% dan MCC 1% pada suhu 60 oC. Tiga isolat (SL1-2-R-2, SL1-2-R-3, dan SL1-2-R-4) yang mendegradasi CMC 1% dan MCC 1% hingga 60 oC merupakan isolat terpilih. Identifikasi dan karakterisasi telah dilakukan pada penelitian sebelumnya dan melaporkan tiga isolat terpilih memiliki kekerabatan terdekat dengan 'Actinomadura keratinilytica' WCC-2665T(=NBRC 105837T). Hasil pengujian menunjukkan 'type strain' NBRC 105837T mendegradasi CMC 1% dan MCC 1% pada medium Mm padat dengan suhu 45, 50, 55, dan 60 oC setelah inkubasi 7 hari. 'Crude enzyme' dari tiga isolat potensial dan 'type strain' NBRC 105837T menunjukkan aktivitas selulolitik pada medium Mm padat yang ditambahkan CMC 1% atau MCC 1% pada suhu 45, 50, 55, dan 60 oC. Analisis filogenetik tiga isolat terpilih berdasarkan gen 16S rRNA menggunakan metode 'Neighbor-Joining' (NJ), 'Minimum Evolution' (ME), dan 'Maximum Likelihood' (ML) menunjukkan bahwa tiga isolat terpilih berada pada satu 'clade' monofiletik dengan 'Actinomadura' 'keratinilytica' WCC-2665T. Analisis filogenetik juga menunjukkan dua kelompok yang terpisah berdasarkan kemampuan menghasilkan selulase pada anggota famili 'Thermomonosporaceae'. ...... The aims of this study were to obtained thermophilic 'Actinobacteria' isolates from soil around Cisolok geyser, West Java with the ability to degrade cellulose at high temperatures and to analyze the phylogenetic position based on 16S rRNA gene of the selected isolates compared to closely related species. Cellulose degradation screening was performed on Minimal (Mm) medium with the addition of 1% (w/v) carboxymethyl cellulose (CMC) or 1% (w/v) microcrystalline cellulose (MCC) as substrate then incubated for 7 days. Cellulose degradations were observed by staining the plates with  0,2% (w/v) Congo red and clear zone formation around the bacterial colony would indicate the cellulose degradation. The results showed that 15 isolates were able to degrade 1% CMC and 12 isolates were able to degrade 1% MCC at 45 oC, 14 isolates were able to degrade 1% CMC and 1% MCC at 50 oC, 4 isolates were able to degrade 1% CMC and 1% MCC at 55 oC, and 3 isolates were able to degrade 1% CMC and 1% MCC at 60 oC. Three isolates (SL1-2-R-2, SL1-2-R-3, and SL1-2-R-4) were selected due to their CMC and MCC degrading ability at 60 oC. Molecular identification based on 16S rRNA gene and characterization in previous study showed that the three selected isolates are closely related to 'Actinomadura keratinilytica' WCC-2665T(=NBRC 105837T). The assay showed that type strain NBRC 105837T was able to degrade 1% CMC and 1% MCC at 45, 50, 55, and 60 oC after 7 days of incubation. Cellulolytic activity show that the crude enzymes of the three selected isolates and type strain were able to degrade 1% CMC and 1% MCC at 45, 50, 55, and 60 oC. Phylogenetic analysis using Neighbour-Joining (NJ), Minimum Evolution (ME), and Maximum Likelihood (ML) methods showed that the  three selected isolates  were  clustered  together in monophyletic clade with 'Actinomadura keratinilytica' WCC-2265T with 100% bootstrap value. Phylogenetic analysis also showed that cellulase  producers  and  non-cellulase  producers  in 'Thermomonosporaceae' were grouped into different clades.
Depok: Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Indonesia, 2020
T54737
UI - Tesis Membership  Universitas Indonesia Library
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Timotius Pramudita Rasendriya
Abstrak :
Rare-actinomycetes tersebar di berbagai habitat terutama di habitat ekstrem seperti kawasan geotermal. Penelitian mengenai rare-actinomycetes dilakukan terkait potensinya sebagai penghasil senyawa bioaktif baru yang bermanfaat dalam bidang kesehatan, industri dan farmasi. Tujuan dari penelitian ini adalah mengisolasi dan mengkarakterisasi secara fenotip dan genotip rare-actinomycetes dari sampel tanah di bawah batuan kuarsa (S3.5.3) di hutan kawasan geotermal Cisolok. Metode pengayaan sampel tanah dilakukan menggunakan medium 10% R2A (Reasoner’s 2A) cair dengan penambahan cycloheximide 100 ppm dan sodium azide 60 ppm, kemudian diinkubasi pada suhu 30°C selama 30 hari. Isolasi rare actinomycetes dilakukan dengan metode membran filter dan spread pada medium 10% R2A gellan gum yang diinkubasi pada suhu 45°C. Karakterisasi dilakukan secara genotipik (berdasarkan data sequence gen 16S rRNA, analisis homologi sequence, dan rekonstruksi pohon filogenetik dengan metode Neighbour-Joining, Maximum Likelihood, dan Minimum Evolution); dan karakterisasi fenotipik (morfologi, fisiologi, dan biokimia). Sebanyak 26 isolat diperoleh dari sampel S3.5.3. Lima isolat dengan karakter morfologi actinomycetes yang diisolasi dari suhu 45°C dengan membran filter, dipilih untuk diidentifikasi. Hasil analisis sequence gen 16S rRNA dari lima isolat menunjukkan persentase homologi sebesar 95,46-99,56% terhadap Micromonospora yasonensis DS3186T. Berdasarkan hasil analisis filogenetik dengan metode Neighbour-Joining, kelima isolat memiliki hubungan kekerabatan terdekat dengan Micromonospora yasonensis DS3186T. Kelima isolat merupakan anggota class Actinomycetes, ordo Micromonosporales, family Micromonosporaceae. Karakter fenotipik kelima isolat sesuai dengan Micromonospora yasonensis sebagai spesies terdekatnya. Kelima isolat merupakan bakteri termotoleran (tumbuh pada suhu 30-45°C dan suhu optimum 40°C), aerobik, Gram positif, menghasilkan miselium substrat tanpa adanya miselium aerial, positif katalase, dan menghasilkan soluble pigment. Penelitian ini mengungkapkan bahwa Micromonospora yasonensis dapat ditemukan di kawasan geotermal dan berasosiasi dengan batuan kuarsa. ......Rare-actinomycetes are distributed in various habitats, particularly in extreme environments such as geothermal areas. Research on rare-actinomycetes focuses on their potential as producers of new bioactive compounds beneficial in health, industrial, and pharmaceutical fields. The aims of this study were to isolate and characterize rare-actinomycetes from soil samples beneath quartz rocks (S3.5.3) in the geothermal forest of Cisolok. Soil sample enrichment was performed using 10% R2A (Reasoner’s 2A) liquid medium supplemented with 100 ppm cycloheximide and 60 ppm sodium azide, incubated at 30°C for 30 days. Rare actinomycetes isolation was carried out using the membrane filter method and spread on 10% R2A agar with gellan gum, incubated at 45°C. Characterization included genotypic analysis based on the sequence of 16S rRNA gene, supported by phenotypic characterization (morphology, physiology, and biochemistry). A total of 26 isolates were obtained from sample S3.5.3. Five isolates with actinomycetes morphology isolated at 45°C using the membrane filter method were selected for characterization. Analysis of the 16S rRNA gene sequences from these five isolates showed homology levels of 95.46-99.56% to Micromonospora yasonensis DS3186T. Based on phylogenetic analysis using the Neighbour-Joining method, the five isolates were most closely related to Micromonospora yasonensis DS3186T. These isolates belong to the class Actinomycetes, order Micromonosporales, and family Micromonosporaceae. Phenotypic characteristics of the five isolates were consistent with Micromonospora yasonensis as their closest species. These isolates are thermotolerant bacteria (growing at temperatures of 30-45°C; optimum temperature 40°C), aerobic, Gram-positive, produce substrate mycelium without aerial mycelium, positive for catalase, and produce soluble pigment. This study reveals that Micromonospora yasonensis can be found in geothermal areas associated with quartz rocks.
Depok: Fakultas Matematika Dan Ilmu Pengetahuan Alam Universitas Indonesia, 2024
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UI - Skripsi Membership  Universitas Indonesia Library
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Dwi Apriyanto
Abstrak :
Penyakit Panama pada tanaman pisang disebabkan oleh kapang patogen Fusarium oxysporum f. sp. cubense. Telah dilakukan penelitian untuk karakterisasi penghambatan Aktinomisetes terhadap Fusarium oxysporum f. sp. cubense secara in vitro menggunakan sel hidup dan filtrat kultur bebas sel. Isolat Aktinomisetes LAI-I dan L31 diketahui menghasilkan enzim kitinase, protease, dan antibiotik yang dapat menghambat pertumbuhan, menyebabkan perubahan morfologi hifa berupa penebalan pada ujung-ujung hifa, serta menghambat germinasi spora Fusarium oxysporum f. sp. cubense. Perhitungan statistik menunjukkan adanya perbedaan yang nyata antara kelompok kontrol, perlakuan LAI-I, dan perlakuan L31. Hasil tersebut memperlihatkan adanya kemampuan isolat LAI-I dan L31 menghambat Fusarium oxysporum f. sp. cubense.
The Panama disease in banana plants is caused by the pathogenic fungi Fusarium oxysporum f. sp. cubense. Research has been conducted to characterize the inhibition mechanism of Actinomycetes towards Fusarium oxysporum f. sp. cubense in vitro by using living cell and cell-free culture filtrate. Actinomycetes isolates LAI-I and L31 produce chitinase enzyme, protease enzyme, and secondary metabolites that can inhibit the growth, lead morphological changes of hyphae as swollen at the end of the hypha, and inhibit spore germination of Fusarium oxysporum f. sp. cubense. The statistic reveals the significant differences between control, LAI-I treatment, and L31 treatment. The result shows the ability of isolate LAI-I and L31 to inhibit Fusarium oxysporum f. sp. cubense.
2014
S53974
UI - Skripsi Membership  Universitas Indonesia Library
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Hana Nadhifah
Abstrak :
Actinomycetes yang diisolasi dari habitat laut diketahui memiliki potensi sebagai penghasil enzim baru yang bermanfaat bagi industri. Kelimpahan keanekaragaman kelompok bakteri ini di Indonesia serta kurangnya penelitian mendorong kebutuhan akan informasi lebih mendalam. Isolat Actinomycetes (BLH 5-14) dari sedimen laut Sarena Kecil, kota Bitung, Sulawesi Utara, menunjukkan potensi sebagai penghasil enzim pektinase dan xilanase. Kedua enzim ini dibutuhkan dalam pengolahan jus buah, kertas, serat rami, dan produksi oligosakarida. Penelitian bertujuan untuk mengkarakterisasi enzim pektinase dan xilanase oleh isolat BLH 5-14 menggunakan substrat komersial. Hasil yang diperoleh menunjukkan bahwa enzim pektinase memiliki aktivitas tertinggi pada hari produksi ke-6 sebesar 4,72±0,08 U/mL dengan pH optimum 8,0 dan suhu optimum 50℃, sedangkan enzim xilanase memiliki aktivitas tertinggi pada hari produksi ke-6 sebesar 7,63±0,03 U/mL dengan pH optimum 6,0 dan suhu optimum 60℃. Inhibitor terbesar bagi kedua enzim merupakan Hg2+ dan SDS, sedangkan ion Fe2+ dapat meningkatkan aktivitas hingga dua kali lipat. Melalui tahapan hidrolisis dan TLC, diketahui pula bahwa enzim pektinase dan xilanase mampu menghasilkan monosakarida seperti galacturonic acid (P1), dan oligosakarida seperti xylotriose (X3), xylotetraose (X4), dan xylopentaose (X5), yang dengan pengembangan lebih lanjut dapat dimanfaatkan sebagai bahan prebiotik bagi industri kesehatan. Berdasarkan hasil sekuens 16S rDNA, isolat Actinomycetes (BLH 5-14) diidentifikasi sebagai genus Streptomyces dengan kemiripan terdekat Streptomyces tendae (99,78%). ......Actinomycetes isolated from marine habitats are known to have the potential for novel enzymes that are beneficial for the industry. The high diversity of the bacterial group in Indonesia and the lack of research that exist have prompted the need for more in-depth information. Actinomycetes isolates (BLH 5-14) that were obtained from marine sediments of Sarena Kecil, Bitung, North Sulawesi, showed potential to produce pectinase and xylanase needed in the processing of fruit juice industry, paper, ramie, and the production of oligosaccharides. This study aimed to characterize the pectinase and xylanase enzymes produced by BLH 5-14 isolates using commercial substrates. The results revealed that the pectinase had the highest activity on the 6th day (4.72±0.08 U/mL) at the optimum pH of 8.0 and optimum temperature of 50℃, while the xylanase had the highest activity on the 6th day (7.63±0.03 U/mL) at optimum pH 6.0 and optimum temperature 60℃. The most significant inhibitors for both enzymes are Hg2+ and SDS, while Fe2+ can increase enzyme activity up to two times the original value. Hydrolysis and thin layer chromatography also showed that pectinase and xylanase were able to produce oligosaccharides such as galacturonic acid (P1), xylotriose (X3), xylotetraose (X4), and xylopentaose (X5), which with further development can be used as prebiotics for the healthcare industry. Based on the results of 16S rDNA sequences, BLH 5-14 were identified as genus Streptomyces, with closely related species Streptomyces tendae (99,78%).
Depok: Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Indonesia, 2022
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UI - Skripsi Membership  Universitas Indonesia Library
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Mazytha Kinanti Rachmania
Abstrak :
Penelitian ini bertujuan untuk memperoleh isolat Actinobacteria termofilik potensial dari tanah di sekitar geiser Cisolok yang dapat mendegradasi xylan dan mengetahui hubungan kekerabatannya dengan taksa terdekat dari Actinobacteria penghasil xylanase. Tujuh belas isolat Actinobacteria termofilik diisolasi dari tanah di sekitar geiser Cisolok, Jawa Barat. Penapisan kemampuan 17 isolat Actinobacteria dan type strain Actinomadura keratinilytica NBRC 105837T mendegradasi xylan dilakukan menggunakan medium Minimal (Mm) padat dengan penambahan substrat xylan 0,5, inkubasi selama 7 hari. Pewarnaan dengan Congo red 0,2 (b/v) menunjukkan terbentuknya zona bening di sekitar koloni isolat Actinobacteria yang dapat mendegradasi xylan 0,5 pada suhu 45 C (15 isolat), 50 C (14 isolat), 55 C (4 isolat), dan 60 C (3 isolat). Type strain NBRC 105837T dapat mendegradasi xylan 0,5 pada suhu 45 C hingga 60 C. Tiga isolat (SL1- 2-R-2, SL1-2-R-3, dan SL1-2-R-4) yang mendegradasi xylan 0,5 hingga suhu 60 C dipilih sebagai isolat potensial. Tiga isolat potensial dan type strain NBRC 105837 dapat mendegradasi substrat Remazol Brilliant Blue R-xylan (RBB-xylan) 0,1 pada medium Mm padat setelah 3 hari inkubasi pada suhu 45 hingga 60 C. Tiga isolat potensial telah diidentifikasi pada penelitian sebelumnya sebagai Actinomadura keratinilytica berdasarkan karakter genotip dan fenotip. Crude enzyme dari tiga isolat potensial dan type strain NBRC 105837 dapat mendegradasi xylan 0,5 dan RBB-xylan 0,1 pada medium Mm padat setelah 24 jam inkubasi pada suhu 45 hingga 60 C. Berdasarkan analisis filogenetik sequence gen 16S rRNA menggunakan metode neighbor-joining, minimum evolution, dan maximum likelihood, 3 isolat potensial membentuk clade yang monofiletik dengan dua spesies Actinomadura termofilik yang dapat mendegradasi xylan (A. keratinilytica dan A. miaoliensis). Tiga isolat potensial membentuk clade yang monofiletik dengan empat spesies Actinomadura termofilik (A. keratinilytica, A. miaoliensis, A. rubrobrunea, dan A. viridilutea). Tiga isolat potensial menghasilkan miselium substrat yang bercabang dan tidak berfragmen, serta miselium aerial yang menghasilkan spora pada medium modified Bennetts padat setelah 14 hari inkubasi pada suhu 45 C. Penelitian ini memberikan informasi tambahan mengenai kemampuan typestrain A. keratinilytica NBRC 105837 mendegradasi xylan.
The aims of this study were to obtain the potential xylan-degrading thermophilic Actinobacteria isolates from soil of Cisolok geysers and to understand their relationship with the closely related taxa of xylanase-producing Actinobacteria. Seventeen thermophilic Actinobacteria isolates were isolated from soil collected around Cisolok geysers, West Java. Xylan-degrading ability of 17 Actinobacteria isolates and type strain Actinomadura keratinilytica NBRC 105837T were screened by using Minimal (Mm) agar medium with the addition of 0.5 xylan substrate, incubated for 7 days. Clear zone was formed around the colony of Actinobacteria isolates which showed xylan-degrading ability at 45 C (15 isolates), 50 C (14 isolates), 55 C (4 isolates), and 60 C (3 isolates) after staining by 0.2 (w/v) Congo red. Type strain NBRC 105837T was able to degrade 0,5 xylan at 45 to 60 C. Three isolates (SL1-2-R-2, SL1-2-R-3, dan SL1-2-R-4) that showed xylan-degrading ability at 45 to 60 C were choosen as potential isolates. Three potential isolates and type strain NBRC 105837T were able to degrade 0,1 Remazol Brilliant Blue R-xylan (RBB-xylan) substrate on Mm agar after 3 days incubation at 45 to 60 C. In the previous study, these potential isolates were identified as Actinomadura keratinilytica based on genotypic and phenotypic characters. Crude enzyme of 3 potential isolates and type strain NBRC 105837T were able to degrade both 0.5 xylan and 0.1 RBB-xylan on Mm agar after 24 hours at 45 to 60 C. Phylogenetic analyses based on 16S rRNA gene using neighbor-joining, minimum evolution, and maximum likelihood methods showed the 3 potential isolates formed monophyletic clade with two thermophilic xylan-degrading Actinobacteria species (A. keratinilytica and A. miaoliensis). Three potential isolates formed monophyletic clade with four thermophilic Actinobacteria species (A. keratinilytica, A. miaoliensis, A. rubrobrunea, and A. viridilutea). These isolates produced non-fragmented branched substrate mycelia and spores produced from aerial mycelia after 14 days incubation at 45 C. This study reports a new information regarding the xylan-degrading ability of A. keratinilytica NBRC 105837.
Depok: Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Indonesia, 2020
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Rifki Alfisyahri
Abstrak :
Telah dilakukan penapisan senyawa antimikroba terhadap 22 isolat Actinomycetes hasil isolasi dari sedimen pesisir Pulau Pramuka, Kepulauan Seribu, DKI Jakarta. Penapisan senyawa antimikroba dilakukan menggunakan metode plug dan hasil penapisan dinyatakan dalam indeks aktivitas IA . Hasil penapisan menunjukkan tidak terdapat aktivitas antimikroba terhadap Escherichia coli NBRC 3301. Namun terdapat 13 isolat yang mampu menghambat pertumbuhan Staphylococcus aureus NBRC 100910 IA 0,461-2,338, sebanyak 19 isolat mampu menghambat pertumbuhan Kocuria rhizophila NBRC 12708 IA 0,705-8,200, ada lima isolat dapat menghambat pertumbuhan Candida albicans UICC Y-29 IA 0,357-0,885, dan terdapat empat isolat yang mampu menghambat pertumbuhan Saccharomyces cerevisiae UICC Y-17 IA 0,357-1,348. Berdasarkan data penapisan, isolat SD 17 ditetapkan sebagai isolat terpilih karena mampu menghambat pertumbuhan dari bakteri Gram positif S. aureus dan K. rhizophila, serta yeast S. cerevisiae dan C. albicans yang diujikan. Penentuan waktu fermentasi yang optimal dari isolat SD17 untuk produksi senyawa antimikroba dilakukan dengan medium CSM Cross Streak Media dan PM4 Production Medium 4 pada hari ke-3, 6, 9, dan 12. Hasil uji aktivitas antimikroba dari filtrat medium pertumbuhan menunjukkan produksi senyawa antimikroba dari isolat SD 17 optimal pada hari ke-9 dengan menggunakan medium CSM. Uji aktivitas antimikroba hasil ekstraksi menggunakan pelarut etil asetat, pada konsentrasi 20 mg/mL, menunjukkan terdapat aktivitas antimikroba terhadap S. aureus IA 2,33, K. rhizhophila IA 4,71, S. cerevisiae IA 1,36 dan Candida albicans IA 0,22. ...... Twenty two isolates of Actinomycetes have been screened for antimicrobial activity, all isolates were isolated from sediment in coastal Pramuka island, Kepulauan Seribu, Jakarta, Indonesia. Strains were screened for antimicrobial activity using plug method and determined by antimicrobial Activity Index AI. The result showed no inhibition activity was observed in the Escherichia coli NBRC 3301. However, there were 13 isolates inhibited Staphylococcus aureus NBRC 100910 0.461 mdash 2.338, 19 isolates inhibited Kocuria rhizophila NBRC 12708 0.705 mdash 8.200, 5 isolates inhibited Candida albicans 0.885 mdash 0.357, and 4 isolates inhibited Saccharomyces cerivisiae 0.357 mdash 1.348. Based on the results of antimicrobial test, SD17 is the most potential strain since it is able to inhibit all Gram positive and yeast tested. To acquire optimal period for antimicrobial fermentation from isolate SD 17, isolates were screened with two different fermentation medium Cross Streak Media CSM and Production Medium 4 PM4. Medium filtrates were tested at 3,6,9 and 12 days incubation. The result showed the optimal activity was observed at 9 days incubation using CSM. The result of antimicrobial test from medium extract with concentration 20 mg mL showed inhibition zone against S. aureus IA 2,33, K. rhizhophila IA 4,71, S. cerevisiae IA 1,36 and Candida albicans IA 0,22.
Depok: Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Indonesia, 2018
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Heru Dwi Prasetiyo
Abstrak :
ABSTRAK
Produksi senyawa antimikroba dari dua Actinomycetes potensial, SRM 2 dan SD 17, telah dilakukan pada medium yang mengandung sumber karbon berupa soluble starch CSMs dan tepung beras CSMb. Uji aktivitas antimikroba dilakukan dengan metode silinder menggunakan filtrat medium pertumbuhan serta hasil ekstraksi senyawa antimikroba. Ekstraksi senyawa antimikroba dari Isolat SRM 2 menggunakan pelarut butanol, sedangkan ekstraksi senyawa antimikroba dari Isolat SD 17 menggunakan pelarut etil asetat. Hasil penelitian menunjukkan Isolat SRM 2 mampu tumbuh lebih baik pada medium CSMs sedangkan Isolat SD 17 mampu tumbuh lebih baik pada medium CSMb. Uji aktivitas antimikroba menunjukkan filtrat medium CSMs dari Isolat SRM 2 mampu menghambat Kocuria rhizophila IA 2,79 0,341, Staphylococcus aureus IA 0,98 0,120, dan Escherichia coli IA 0,92 0,209. Sementara itu, filtrat medium CSMb dari Isolat SRM 2 mampu menghambat K. rhizophila IA 2,66 0,279, S. aureus IA 0,97 0,115, dan E. coli IA 0,89 0,139. Filtrat medium CSMs dari Isolat SD 17 mampu menghambat K. rhizophila IA 1,75 0,270, S. aureus IA 1,41 0,119, Candida albicans IA 0,67 0,104, dan Saccharomyces cerevisiae IA 0,93 0,247. Sementara itu, filtrat medium CSMb dari Isolat SD 17 mampu menghambat K. rhizophila IA 3,16 0,085, S. aureus IA 3,05 0,256, C. albicans IA 0,95 0,256, dan S. cerevisiae IA 1,30 0,110. Hasil ekstraksi senyawa antimikroba Isolat SD 17 dari medium CSMb menunjukkan produktivitas lebih tinggi dibandingkan medium CSMs. Ekstrak dari medium CSMs mampu menghambat K. rhizophila IA 3,76 0,179, S. aureus IA 0,61 0,105, dan C. albicans IA 1,37 0,176. Sementara itu, ekstrak dari medium CSMb mampu menghambat K. rhizophila IA 3,21 0,119, S. aureus IA 0,61 0,082, dan C. albicans IA 2,35 0,111. Hasil ekstraksi senyawa metabolit sekunder Isolat SRM 2 dari medium CSMs menunjukkan produktivitas lebih tinggi dibandingkan medium CSMb. Namun kedua ekstrak tidak memiliki aktivitas antimikroba.
ABSTRACTbr> Production of antimicrobial compounds from two potential Actinomycetes, SRM 2 and SD 17, were performed on medium containing soluble starch CSMs and rice flour CSMb as a carbon source. The antimicrobial activity test was performed using cylinder method on the growth medium filtrate and crude extract of antimicrobial compound. The extraction of antimicrobial compounds of the SRM 2 used butanol as a solvent, while the extraction of the antimicrobial compounds of the SD 17 used ethyl acetate. The results showed that Isolate SRM 2 was able to grow better in CSMs medium, while Isolate SD 17 was able to grow better in CSMb medium. The antimicrobial activity test showed that CSMs medium filtrate of SRM 2 was able to inhibit Kocuria rhizophila IA 2,79 0.341, Staphylococcus aureus IA 0.98 0.120, and Escherichia coli IA 0.92 0.209. The CSMb medium filtrate of SRM 2 was able to inhibit K. rhizophila IA 2.66 0.279, S. aureus IA 0.97 0.115, and E. coli IA 0.89 0.139. The CSMs medium filtrate of SD 17 was able to inhibit K. rhizophila IA 1.75 0.270, S. aureus IA 1.41 0.119, Candida albicans IA 0.67 0.104, and Saccharomyces cerevisiae IA 0, 93 0.247. The CSMb medium filtrate of SD 17 was able to inhibit K. rhizophila IA 3,16 0.085, S. aureus IA 3.05 0.256, C. albicans IA 0.95 0.256, and S. cerevisiae IA 1.30 0.110. The crude extract of SD 17 from CSMb medium showed higher productivity than CSMs medium. Crude extract from CSMs medium was able to inhibit K. rhizophila IA 3.76 0.179, S. aureus IA 0.61 0.105, and C. albicans IA 1.37 0.176. Meanwhile, crude extract of CSMb medium was able to inhibit K. rhizophila IA 3.21 0.119, S. aureus IA 0.61 0.082 , and C. albicans IA 2.35 0.111. The crude extract of SRM 2 from CSMs medium showed higher productivity than CSMb medium. However both extracts did not display antimicrobial activity.
2018
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