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"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.

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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."

"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."

"Telah dilakukan penelitian yang bertujuan untuk mengetahui aktivitas antimikroba actinomycetes termofil hasil isolasi dari geiser di Cisolok, Jawa Barat. Delapan belas isolat yang memiliki morfologi menyerupai actinomycetes berhasil diisolasi dari serasah daun dan ranting di sekitar pusat semburan geiser. Seluruh isolat diuji aktivitas antimikrobanya menggunakan paper disk method dan agar block method dengan Kocuria rhizophila, Staphylococcus aureus, Bacillus subtilis sebagai bakteri uji Gram positif, dan Escherichia coli sebagai bakteri uji Gram negatif. Pengujian menggunakan metode paper disk menunjukkan hasil negatif pada isolat actinomycetes yang dikultur pada medium International Streptomyces Project (ISP) 1 cair selama 14 hari pada suhu 50oC dan 40oC. Berdasarkan uji menggunakan metode blok agar, didapatkan bahwa dua isolat, yaitu LC2-2 dan LC2-6 memberikan hasil positif terhadap bakteri uji Gram positif. Isolat LC2-2 menunjukkan morfologi makroskopis dan mikroskopis menyerupai genus Bacillus sehingga tidak digunakan untuk identifikasi molekuler. Hasil identifikasi molekuler sequence parsial gen 16S rRNA menggunakan primer 785F dan primer 802R menunjukkan bahwa LC2-6 diidentifikasi sebagai Actinomadura keratinilyitica dengan nilai homologi 99%. Berdasarkan hasil penelitian, direkomendasikan untuk mempelajari lebih lanjut senyawa antimikroba yang dihasilkan isolat LC2-6. Hal tersebut disebabkan oleh belum adanya laporan penelitian mengenai aktivitas antimikroba Actinomadura keratinilytica. ......The aim of this study was to screen the antimicrobial activity by actinomycetes isolated from Cisolok Geyser, West Java. Eighteen isolates which are have similar morphology with actinomycetes have been isolated from leaves and branches around the geyser. The isolates were screened for their antimicrobial activity using paper disk method and agar block method with Kocuria rhizophila, Staphylococcus aureus, Bacillus subtilis as Gram positive test bacteria and Escherichia coli as Gram negative test bacteria. Screening by paper disk method showed negative result from all the isolates that cultured on International Streptomyces Project (ISP) 1 medium at 50oC and 40oC for 14 days. Screening by block agar method showed that two isolates, LC2-2 and LC2-6 gave positive result to Gram positive test bacteria. Morphologically, LC2-2 showed similarity to genus Bacillus, thus it’s not used for molecular identification. Molecular identification based on partial sequence of 16S rRNA gene with primer 785F and primer 802R showed that LC2-6 identified as Actinomadura keratinilytica (99%). Based on this research, it is suggested to do further study about the antimicrobial activity produced by LC2-6, because there is still no report about antimicrobial activity produced by Actinomadura keratinilytica."

"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.'"