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"Penelitian bertujuan memperoleh identitas isolat-isolat actinomycetes dari Raja Ampat, Papua penghasil antimikroba dan antikanker dengan aktivitas tertinggi. Penapisan aktivitas antimikroba dilakukan menggunakan metode difusi agar terhadap bakteri (Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Micrococcus luteus) dan khamir (Candida albicans dan Saccharomyces cerevisiae). Metabolit dari isolat terpilih yang memiliki aktivitas antimikroba tertinggi dianalisis lanjut. Nilai Minimum inhibitor concentration (MIC) dari senyawa murni yang diproduksi isolat terpilih terhadap mikroba diukur menggunakan metode mikrodilusi. Kebocoran asam nukleat dan protein dari mikroba uji dideteksi dengan menggunakan metode spektrofotometri pada panjang gelombang 260 dan 280 nm. Kebocoran urasil dianalisis menggunakan HPLC. Perubahan morfologi diamati menggunakan scanning electron microscope (SEM). Uji sitotoksik dilakukan terhadap 6 sel kanker (MCM-B2, Leukemia, T47D, HeLa, A549, dan WiDr) dan satu sel normal (Vero) menggunakan metode trypan blue dan MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide). Sebanyak sembilan isolat potensial penghasil antimikroba tertinggi (BL-36-1, BL-20-2, BL-14-2, RC-SS-37-4, RC-SS-37-16, BL-22-1, BL-06-5, BL-22-3, dan BL-22-5) diidentifikasi berdasarkan data sekuen penuh gen 16S rRNA (± 1.500 PB). Hasil penelitian menunjukkan 44 % isolat actinomycetes memiliki aktivitas antimikroba. Senyawa F.5.1 merupakan senyawa murni yang diproduksi oleh isolat terpilih BL-22-5. Nilai MIC senyawa tersebut terhadap mikroba uji berkisar 16 -- 64 µg/ml. Aktivitas antifungi senyawa F.5.1 lebih tinggi dibandingkan antibakteri. Senyawa F.5.1 menyebabkan kebocoran protein, asam nukleat, dan urasil pada mikroba uji. Nilai IC50 senyawa F.5.1 berkisar 0,02 -- 3,81 µg/ml terhadap sel kanker dan ≥ 30.000 µg/ml untuk sel normal. Isolat RCSS-37-4, RC-SS-37-16, BL-22-3, dan BL-22-5 teridentifikasi sebagai Streptomyces costaricanus (100 %), Streptomyces costaricanus (99,8 %), Streptomyces parvulus (98,6 %), dan Streptomyces badius (98,9 %). Lima isolat teridentifikasi sebagai Streptomyces spp. (BL-36-1, BL-20-2, BL-14-2, BL-22-1 dan BL-06-5) dan menjadi kandidat spesies baru karena memiliki nilai homologi yang rendah terhadap spesies terdekatnya (93,9% -- 97,4 %).

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The objective of this research was to obtain the identity of actinomycetes isolates from Raja Ampat, Papua-producing antimicrobial and anticancer with the highest activity. Antimicrobial screening was conducted based on agar diffusion method against bacteria (Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Micrococcus luteus) and yeast (Candida albicans and Saccharomyces cerevisiae). Metabolite from selected isolates with the highest antimicrobial activity was chosen for subsequent analysis. Minimum inhibitor concentration (MIC) value against microbial tested of pure bioactive compound was determined using microdilution method. Leakages of nucleic acids and proteins from microbial tested were detected using UV-vis spectrophotometer method on 260 and 280 nm wavelengths. Uracil Leakage was analyzed using HPLC. Morphological changes were observed using a scanning electron microscope (SEM). Determination of cytotoxic activity was conducted in 6 cell lines (MCM-B2, Leukemia, T47D, HeLa, A549, and WiDr) and normal cell (Vero) using trypan blue and MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) methods. Nine selected isolates with the highest antimicrobial activity (BL-36-1, BL-20-2, BL-14-2, RC-SS-37-4, RC-SS-37-16, BL-22-1, BL- 06-5, 22-3-BL and BL-22-5) were identified based on full sequence of 16S rRNA gene (1,500 bp). The results showed 44% of isolates of actinomycetes have antimicrobial activity. F.5.1 is a compound produced by the selected isolate (BL-22-5). MIC values of this compound against tested microbes in range of 16 -- 64 µg/ml. Antifungal activity from F.5.1 compound was higher than antibacterial activity. F.5.1 compound caused leakage of proteins, nucleic acids, and uracil on tested microbes. Inhibitor concentration 50% (IC50) value in the range of 0.02 -- 3.81 µg/ml against cancer cells and ≥ 30,000 µg/ml for normal cells. Isolate RC-SS-37-4, RC-SS-37-16, BL-22-3, and BL-22-5 have been identified as Streptomyces costaricanus (100%), Streptomyces costaricanus (99.8%), Streptomyces parvulus ( 98.6%), and Streptomyces badius (98.9%), respectively. Five isolates (BL-36-1, BL-20-2, BL-14-2, BL-22-1 and BL-06-5) were identified as Streptomyces spp. and can be presumed as candidates for new species because of the low homology value to their closest related species (93.9 % -- 97.4%)."

"Albertisia papuana Becc termasuk tumbuhan tropis dari famili Menispermaceae. Tumbuhan ini dikenal berkhasiat obat, diantaranya sebagai antibiotik/antibakteri. Selain tumbuhan, mikroorganisme termasuk jamur endofit juga dapat menghasilkan antibiotik. Jamur endofit termasuk mikroorganisme yang hidup pada tumbuhan inangnya. Jamur endofit di alam jumlahnya melimpah (1,5 juta dibandingkan tumbuhan sekitar 300 ribuan). Jamur endofit dapat memproduksi metabolit bioaktif yang beragam. Di lain sisi, jamur endofit belum tereksplorasi secara maksimal. Penelitian ini dilakukan untuk menskrining dan mengisolasi senyawa bioaktif dari jamur endofit dari tumbuhan A. papuana sebagai antibiotik. Dari kegiatan penelitian didapatkan 15 isolat jamur endofit yaitu dari bagian batang 8 isolat dan daun 7 isolat. Dari skrining aktivitas antibakteri dengan metode TLC bioassay didapatkan informasi 2 isolat jamur endofit yang bersifat paling aktif yaitu DAP KRI-5 dan BAP KRI-8. Dari pemisahan dan pemurnian didapatkan 2 buah senyawa murni dari DAP KRI-5 yaitu F4.3, dan F2.3.9. Hasil dari elusidasi struktur menggunakan spektr. 1H dan 13C-NMR; UV-Vis; dan GC-MS menunjukkan F4.3 adalah C6H6O3 yaitu floroglusinol. Floroglusinol mempunyai aktivitas antibakteri melawan S. aureus sama kuatnya dengan klorampenikol dengan nilai MIC yaitu 64 𝜇g/mL, namun sampel F4.3 bersifat parsial sebagai antibakteri. Berdasarkan spektr. 1H dan 13C-NMR, 2D NMR dengan DEPT; HMBC; HMQC; dan 1H-1H COSY, spektr. UV-Vis dan IR, dan ToF ESI-MS menunjukkan F2.3.9 mempunyai rumus molekul C30H37NO6 yaitu sitokalasin D.

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Albertisia papuana Becc is tropical plants that belong to the family of Menispermaceae. It was known as medicine, such as an antibiotic/antibacteria. Besides plants, microorganisms including endophytic fungi also can produce antibiotics. Endophytic fungi live in their host plant. Endophytic fungi have abundant number in the world (1.5 million compared to approximately 300 thousands of plant). They can produce diversity of bioactive metabolites. The other hand, they have not been maximized exploration yet. This study was conducted for screening and isolating of bioactive compounds of endophytic fungi from A. papuana as antibiotics. This research activities obtained 15 isolates of the endophytic fungi. The isolates are from the stem and leaf, 8 and 7 isolates respectively.

Screening of antibacterial activity with TLC bioassay obtained two isolates which have the most active as antibacterial, there are DAP KRI-5 and BAP KRI-8. Separation and purification obtained two pure compounds from KRI DAP-5, there are F4.3, and F2.3.9. The results of structure elucidation by spectr. 1H and13C-NMR, UV-Vis, and GC-MS showed F4.3 is C6H6O3, phloroglucinol. Phloroglucinol has antibacterial activity against S. aureus as well as chloramphenicol with MIC value are 64 𝜇g/mL, but F4.3 partially activity as antibacterial agent. Based on spectr. 1H and 13C-NMR, 2D NMR with DEPT; HMBC; HMQC; and 1H-1H COSY, spectr. UV-Vis and IR, and ToF ESI-MS showed F2.3.9 is C30H37NO6, cytochalasin D."