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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 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-2665^T(=NBRC 105837^T). Hasil pengujian menunjukkan 'type strain' NBRC 105837^T 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 105837^T 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-2665^T. Analisis filogenetik juga menunjukkan dua kelompok yang terpisah berdasarkan kemampuan menghasilkan selulase pada anggota famili 'Thermomonosporaceae'.

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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-2665^T(=NBRC 105837^T). The assay showed that type strain NBRC 105837^T 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-2265^T with 100% bootstrap value. Phylogenetic analysis also showed that cellulase  producers  and  non-cellulase  producers  in 'Thermomonosporaceae' were grouped into different clades."

"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.

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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 memperoleh identitas dua isolat bakteri termofilik dari geiser. Isolat LC2-23 diperoleh dari serasah pada geiser di Cisolok, Jawa Barat, Indonesia, dan isolat RKB-2 diperoleh dari serasah pada geiser di Onikobe, Miyagi, Jepang.!!Identifikasi dilakukan berdasarkan gabungan data fenotipik dan genotipik. Berdasarkan karakterisasi fenotipik, isolat LC2-23 memiliki sel berbentuk batang; menghasilkan endospora; motil; gram positif; bersifat aerob dan fakultatif aerob; mampu tumbuh pada suhu 60 oC, sedangkan suhu optimum pertumbuhan 50 oC. Berdasarkan karakterisasi genotipik, data full sequence gen 16S rRNA isolat LC2-23 memiliki homologi 99,1% terhadap Brevibacillus agri. Berdasarkan data fenotipik dan genotipik, isolat LC2-23 diidentifikasi sebagai Brevibacillus agri (Family Paenibacillaceae, Order Bacillales, Class Bacilli, Phylum Firmicutes). Berdasarkan karakterisasi fenotipik, isolat RKB-2 membentuk miselium vegetatif dan aerial yang bercabang; menghasilkan spora aerial; gram positif; bersifat aerob; mampu tumbuh pada suhu 60 oC, sedangkan suhu optimum pertumbuhan 50 oC. Berdasarkan karakterisasi genotipik, data full sequence gen 16S rRNA isolat RKB-2 memiliki homologi yang rendah, yaitu 98,4% terhadap spesies terdekatnya, Thermosporothrix hazakensis (Family Thermosporotrichaceae, Order Ktedonobacteriales, Class Ktedonobacteria, Phylum Chloroflexi). Hasil analisis filogenetik menunjukkan posisi isolat RKB-2 terpisah dari T. hazakensis. Data kemotaksonomi (komposisi asam lemak) dan hasil analisis proteomik menggunakan MALDI-TOF MS mendukung perbedaan antara isolat RKB-2 dan T. hazakensis. Berdasarkan perbedaan tersebut isolat RKB-2 diidentifikasi sebagai spesies baru dari Thermosporothrix. Untuk pengajuan nama spesies baru diperlukan data hibridisasi DNA-DNA antara isolat RKB-2 dengan T. hazakensis.

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This research was aimed to identify two bacterial isolates obtained from geysers. Strain LC2-23 was isolated from litters on a geyser in Cisolok, West Java, Indonesia, and isolate RKB-2 was obtained from litters on a geyser in Onikobe, Miyagi Prefecture, Japan. Identification of bacteria was based on integrated data of phenotypic and genotypic characterizations. Based on phenotypic characterizations of isolate LC2-23: it has a rod (bacilli)-shaped cells, forms endospores; gram positive; motile; aerobic, and able to grow up to a temperature of 60 oC. Based on genotypic characterizations of isolate LC2-23: the full sequence of genes 16S rRNA shows 99.1% sequence homology to Brevibacillus agri. Based on phenotypic and genotypic data, isolate LC2-23 can be identified as Brevibacillus agri (Family Paenibacillaceae, Order Bacillales, Class Bacilli, Phylum Firmicutes). Based on phenotypic characterizations of isolate RKB-2: vegetative and branching aerial mycelia forms, gram positive, aerobic, and able to grow up to a temperature of 60 oC. Based on genotypic characterizations of isolate RKB-2: the full sequence of 16S rRNA gene of isolate RKB-2 showed low homology (98.4%) to Thermosporothrix hazakensis (Family Thermosporotrichaceae, Order Ktedonobacteriales, Class Ktedonobacteria, Phylum Chloroflexi). Phylogenetic analysis showed the isolate RKB-2 was distinct from cluster of Thermosporothrix hazakensis and Ktedonobacteria bacterium. The genotypic and phylogenetic data, plus chemotaxonomic and proteomic analysis using MALDI-TOF MS, suggest that isolate RKB-2 represent novel species of the genus Thermosporothrix. The DNA-DNA hibridization data is needed for proposal of new species."

"Penelitian ini bertujuan untuk memperoleh informasi mengenai pengaruh variasi mediumpertumbuhan terhadap pembentukan miselium aerial dan aktivitas antimikroba delapanisolat rare Actinobacteria termofilik dari tanah di sekitar geiser Cisolok, Jawa Barat.Pengujian pertumbuhan, pembentukan miselium aerial, dan aktivitas antimikrobadilakukan dengan menumbuhkan isolat pada medium ISP 1 agar, ISP 1 gellan gum, ISP2 agar, ISP 2 gellan gum, ISP 3 agar, ISP 3 gellan gum, Bennett’s agar, Bennett’s gellangum, Minimal (Mm) 3 agar, Mm 3 gellan gum, 2% agar, dan 2% gellan gum. Isolatkemudian diinkubasi pada suhu 45 °C selama 7 dan 14 hari. Konfirmasi suhupertumbuhan menunjukkan 2 isolat dapat tumbuh hingga suhu 45 °C dan 6 isolat dapattumbuh hingga 50 °C. Hasil pengujian variasi medium pertumbuhan menunjukkan semuaisolat rare Actinobacteria dapat menghasilkan miselium substrat pada semua medium.Hasil pengamatan setelah inkubasi selama 7 hari pada suhu inkubasi 45 °C menunjukkanisolat-isolat tersebut dapat menghasilkan miselium aerial pada medium ISP 1 agar (2isolat), Mm 3 agar (3 isolat), 2% agar (5 isolat), dan 2% gellan gum (5 isolat). Hasilpengamatan setelah inkubasi selama 14 hari menunjukkan isolat-isolat tersebut dapatmenghasilkan miselium aerial pada medium ISP 1 gellan gum (2 isolat), ISP 2 agar (1isolat), ISP 2 gellan gum (3 isolat), ISP 3 agar dan gellan gum (2 isolat), Mm 3 agar 3isolat, dan Mm 3 gellan gum (3 isolat). Hasil pengujian aktivitas antibakteri menunjukkanisolat SL3-2-R-11 yang ditumbuhkan pada medium ISP 3 gellan gum dan SL3-1-R-7yang ditumbuhkan pada Bennett’s agar selama 7 hari dapat menghambat pertumbuhanStaphylococcus aureus. Hasil pengujian aktivitas antikhamir menunjukkan isolat SL3-2-R-11 yang ditumbuhkan pada medium ISP 3 gellan gum dan SL3-1-R-7 pada mediumBennett’s agar selama 14 hari dapat menghambat pertumbuhan Candida albicans. Hasilpengujian aktivitas antifungi menunjukkan tidak ada isolat yang dapat menghambatpertumbuhan Aspergillus flavus.

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This study aims to obtain information about the effect of growth medium variations onthe formation of aerial mycelium and antimicrobial activity of eight thermophilic rareActinobacteria isolates from the soil around Cisolok geyser, West Java. The ability togrow at various media, aerial mycelium formation, and antimicrobial activity were carriedout by growing isolates on medium ISP 1 agar, ISP 1 gellan gum, ISP 2 agar, ISP 2 gellangum, ISP 3 agar, ISP 3 gellan gum, Bennett’s agar, Bennett’s gellan gum, Minimum (Mm)3 agar, Mm 3 gellan gum, 2% agar, and 2% gellan gum. The isolates were then incubatedat 45 oC for 7 and 14 days. Growth test at various temperatures showed that two isolatescould grow at a temperature of 45 oC and six isolates could grow up to 50 oC. The resultsof the growth medium variation test showed that all rare Actinobacteria isolates couldproduce substrate mycelium in all mediums. Observations after incubation for 7 days at45 °C showed that these isolates could produce aerial mycelium on ISP 1 agar medium(2 isolates), Mm 3 agar (3 isolates), 2% agar (5 isolates), and 2% gellan gum (5 isolates).Observations after incubation for 14 days showed that these isolates could produce aerialmycelium on the medium ISP 1 gellan gum (2 isolates), ISP 2 agar (1 isolate), ISP 2gellan gum (3 isolates), ISP 3 agar and gellan gum (2 isolates), Mm 3 agar 3 isolates, andMm 3 gellan gum (3 isolates). The results of antibacterial activity test showed that isolatesSL3-2-R-11 grown on ISP 3 gellan gum and SL3-1-R-7 grown on Bennett’s agar for 7days could inhibit the growth of Staphylococcus aureus. The antifungal activity test ofisolates SL3-2-R-11 grown on ISP 3 gellan gum medium and SL3-1-R-7 on Bennett’sagar for 14 days showed inhibition towards Candida albicans. Meanwhile, all isolates didnot show antifungal activity against Aspergillus flavus"

"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'.

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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.'"

"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.

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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."

"Kelompok Actinobacteria berfilamen merupakan bakteri Gram positif yang beberapa anggotanya diketahui memiliki kemampuan mendegradasi selulosa dengan menghasilkan selulase. Penelitian ini bertujuan untuk mengetahui kemampuan tumbuh isolat Actinobacteria-like GL1-2, GL1-9, dan GL1-12 pada variasi media agar (ISP 1, ISP 2, ISP 3, dan modified Bennett’s) dan suhu (25, 30, 35, 40, 45, 50, dan 55°C), serta mengetahui kemampuan selulolitiknya pada substrat 1% CMC di berbagai suhu (30, 35, 40, 45, 50, dan 55°C). Kemampuan selulolitik diuji dengan menginokulasi biakan pada medium agar minimal (Mm) dengan penambahan 1% CMC, kemudian diinkubasi pada berbagai suhu selama 3, 7, dan 14 hari. Kemampuan selulolitik diamati dengan terbentuknya zona bening di sekitar koloni setelah ditetesi 0,1% Congo red dan dibilas dengan larutan NaCl 1 M. Isolat GL1-2 dan GL1-9 menunjukkan pertumbuhan miselium substrat dalam jumlah banyak pada semua medium yang diuji, namun sporulasi penuh hanya teramati pada medium ISP 1 agar dan MBA. Isolat GL1-12 menunjukkan pertumbuhan miselium substrat yang baik kecuali pada medium ISP 2 agar, namun sporulasi hanya teramati pada medium ISP 3 agar. Suhu pertumbuhan isolat GL1-2 dan GL1-9 berkisar antara 30--55°C, sedangkan GL1-12 berkisar antara 35--55°C. Hasil uji kemampuan selulolitik menunjukkan bahwa isolat GL1-2 dan GL1- 9 memiliki kemampuan mendegradasi 1% CMC pada suhu 30, 35, 40, 45, 50, dan 55°C. Isolat GL1-12 memiliki kemampuan selulolitik pada suhu 40, 45, 50, dan 55°C. Hasil penelitian menunjukkan bahwa ketiga isolat Actinobacteria-like dari serasah di kawasan sumber air panas gunung Galunggung memiliki potensi menghasilkan enzim selulase di berbagai suhu yang diuji.

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Members of Gram-positive filamentous Actinobacteria are some recognized for their ability to degrade cellulose by producing cellulase. This study aimed to determine the growth ability of three Actinobacteria-like isolates (designated isolates GL1-2, GL1- 9, and GL1-12) obtained from litter samples of mount Galunggung hot spring, Tasikmalaya, West Java, on various agar media (ISP 1, ISP 2, ISP 3, and modified Bennett’s) and temperatures (25, 30, 35, 40, 45, 50, 55°C), along with their cellulolytic ability on 1% carboxymethyl cellulose (CMC) as substrate. Cellulolytic ability was tested by inoculating the cultures on minimal (Mm) agar plates with the addition of 1% CMC, and incubated at various temperatures (30, 35, 40, 45, 50, and 55°C) for 3, 7, and 14-days. Cellulolytic ability was observed as formation of clear zone surrounding the colonies after being flooded with 0.1% Congo red and rinsed with 1 M NaCl solution. The results showed that isolates GL1-2 and GL1-9 have abundant substrate mycelia formation on all media tested, while optimal sporulation was only observed on ISP 1 agar and MBA. Isolate GL1-12 showed good growth of substrate mycelia except on ISP 2 agar, however sporulation was poorly observed only on ISP 3 agar. Growth temperatures of isolates GL1-2 and GL1-9 were ranging from 30 to 55°C, while GL1- 12 was ranging from 35 to 55°C. Isolates GL1-2 and GL1-9 have the ability to degrade 1% CMC at 30, 35, 40, 45, 50, and 55°C. Isolate GL1-12 has celulolytic ability at temperatures of 40, 45, 50, and 55°C. This study revealed that Actinobacteria-like isolates obtained from litter samples of mount Galunggung hot spring, Tasikmalaya are potential cellulase-producers on various tested temperatures."

"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."

"DNA polimerase merupakan sebuah enzim yang memiliki aplikasi luas dalam dunia bioteknologi. Peran DNA polimerase dalam sintesis DNA membuat DNA polimerase sebuah reagen yang sering digunakan pada berbagai metode amplifikasi DNA. Proses sintesis DNA pada metode-metode tersebut menggunakan suhu tinggi, sehingga dibutuhkan DNA polimerase yang tidak terdenaturasi pada suhu tinggi atau dengan kata lain termostabil. Salah satu sumber DNA polimerase termostabil adalah bakteri termofilik Geobacillus thermoleovorans Strain SGAir0734 (GBK Pol) yang diisolasi dari mata air panas Batu Kuwung, Banten, Indonesia. Pada penelitian sebelumnya, telah diuji coba produksi dari GBK Pol dengan suhu purifikasi pada rentang 60 – 80℃ dimana diduga terjadi denaturasi dari enzim. Maka dari itu, pada penelitian ini dilakukan purifikasi GBK Pol sekuens utuh dan parsial dengan metode immobilized metal affinity chromatography (IMAC) dengan rentang suhu 40 – 60℃ dan uji aktivitas amplifikasi DNA dengan metode loop mediated isothermal amplification (LAMP) dengan rentang suhu 25 – 60℃. Hasil uji Lowry menunjukkan bahwa pemanasan 60℃ menghasilkan GBK pol dengan konsentrasi tertinggi, yaitu sekitar 134 µg/ml untuk plasmid sekuens utuh. GBK pol hasil purifikasi kemudian diuji aktivitasnya dengan LAMP, dimana reaksi pada suhu 40℃ selama 2 jam menghasilkan pita yang jelas pada elektroforesis dan menghasilkan konsentrasi DNA tertinggi. GBK pol dari hasil purifikasi dengan pemanasan 60℃ menghasilkan konsentrasi DNA tertinggi pada LAMP suhu 40℃ baik untuk sekuens parsial dan utuh.

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DNA polymerase is an enzyme that has a wide application in the world of biotechnology. DNA Polymerase’s role in synthesizing DNA makes DNA polymerase a favorable reagent in various DNA amplification methods. The synthesis process in these methods is done in high temperatures, thus a DNA polymerase that won’t experience denaturation in high temperatures, or in other words thermostable, is needed. One source of thermostable DNA polymerase is the thermophilic bacteria Geobacillus thermoleovorans Strain SGAir0734 (dubbed GBK Pol) that has been isolated from Batu Kuwung hot springs, Banten, Indonesia. In the previous study, a production trial of GBK pol has been carried out with purification temperatures that range between 60 – 80℃, where denaturation of the enzyme is suspected to have happened. As such, in this study, lower temperatures of 40 – 60℃ are employed in the purification of GBK Pol with immobilized metal affinity chromatography (IMAC) as well as in the activity study with loop mediated isothermal amplification (LAMP) with reaction temperatures of 25 – 60℃. The Lowry assay results show that the highest GBK pol concentration is achieved with 60℃ heat treatment, with a concentration of around 134 µg/ml for full sequence plasmid. Purified GBK pol are then tested with LAMP, where isothermal amplification at 40℃ for 2 hours resulted in the clearest bands during gel electrophoresis as well as highest concentrations of DNA. The highest concentration of DNA is achieved from GBK pol with 60℃ heat treatment, suggesting that 60℃ is the optimal temperature for purification."