http://repository.ipb.ac.id/handle/123456789/53 Wed, 08 Jul 2026 07:14:30 GMT 2026-07-08T07:14:30Z http://repository.ipb.ac.id/handle/123456789/174171 Optimasi Metode Isolasi Gen amyS Penyandi a-Amilase dari Bacillus paralicheniformis ATCC 12759 FITRIANI, TASYA Bakteri pembentuk endospora dikenal sebagai penghasil enzim termostabil, termasuk a-amilase termostabil yang banyak digunakan dalam industri berbasis pati. Sebanyak 60% produksi a-amilase dihasilkan oleh genus Bacillus. Namun, penggunaan bakteri native dalam industri membutuhkan energi besar, sehingga diperlukan teknologi DNA rekombinan. Penelitian ini bertujuan mengoptimasi proses isolasi gen meliputi isolasi DNA genom hingga konfirmasi urutan nukleotida gen amyS dari B. paralicheniformis ATCC 12759 melalui sekuensing Sanger. Gen a-amilase diisolasi menggunakan metode PCR dengan sepasang primer spesifik, yaitu amyS_F1 (5`-ATGAAACAACACAAACGGC-3`) dan amyS_R1 (5`-CTATCTTTGAACATAGATCGAAACC-3`). DNA genom menunjukkan hasil optimum setelah penambahan volume proteinase K, tanpa inkubasi setelah isopropanol, dan buffer TE bersuhu 25 ? untuk resuspensi DNA. Sementara itu, isolasi gen amyS tervisualisasikan dengan pita tunggal ukuran sekitar 1,5 kb pada konsentrasi cetakan DNA 25 ng dan suhu annealing 54,7 ?. Konfirmasi hasil amplifikasi gen menunjukkan urutan nukleotida memiliki tingkat kesamaan 100% dengan gen amyS referensi.; Endospore-forming bacteria are recognized as producers of thermostable enzymes, including thermostable a-amylase which is widely used in starch-based industries. Approximately 60% of a-amylase production is derived from the genus Bacillus. However, the use of native bacteria in industrial applications requires high energy consumption; therefore, recombinant DNA technology is necessary. This study aimed to optimize the gene isolation process, from genomic DNA extraction to nucleotide sequence confirmation of the amyS gene from B. paralicheniformis ATCC 12759 through Sanger sequencing. The a-amylase gene was isolated using the PCR method with a pair of specific primers, namely amyS_F1 (5`-ATGAAACAACACAAACGGC-3`) and amyS_R1 (5`-CTATCTTGAACATAGATCGAAACC-3`). Genomic DNA yielded optimum results following the addition of proteinase K, precipitation with cold isopropanol without an incubation, and dissolution using TE buffer at 25 ?. Meanwhile, isolation of the amyS gene was visualized as a single band of approximately 1,5 kb at a DNA template concentration 25 ng and an annealing temperature of 54,7 ?. Confirmation of the gene amplification product revealed that the nucleotide sequence exhibited 100% identity to the reference amyS gene sequence. Thu, 01 Jan 2026 00:00:00 GMT http://repository.ipb.ac.id/handle/123456789/174171 2026-01-01T00:00:00Z http://repository.ipb.ac.id/handle/123456789/174105 Identifikasi dan Interaksi Molekuler Senyawa Bioaktif Buah Pare (Momordica charantia L.) terhadap Reseptor Xantin Oksidase In Silico Mediartha, I Ketut Hiperurisemia dipicu oleh aktivitas berlebih dari enzim xantin oksidase menyebabkan penyakit kardiovaskular yang parah. Ekstrak etanol dan air buah pare (Momordica charantia L.) diketahui mampu menekan sintesis asam urat, namun mekanisme molekulernya masih belum diteliti lebih lanjut. Penelitian ini bertujuan mengidentifikasi senyawa bioaktif dari buah pare (NusaFarm) dan mengevaluasi interaksi molekulernya terhadap reseptor xantin oksidase (PDB ID: 1FIQ) secara in silico. Metode maserasi menggunakan etanol 96% dan analisis LC-HRMS berhasil menganotasi 132 senyawa yang didominasi oleh asam karboksilat, tanpa adanya senyawa penciri utama. Senyawa penciri minor yang terdeteksi meliputi coumarin dan benzaldehyde. Uji bioavailabilitas, ADME, prediksi toksisitas, serta penapisan virtual (?Gbind and Kd) dievaluasi dengan membandingkannya terhadap asam salisilat dan allopurinol. Hasil penapisan menunjukkan tiga senyawa tidak toksik, namun senyawa dengan interaksi molekuler dan afinitas terbaik coumarin tetap divisualisasikan sebagai senyawa penuntun. Senyawa 4-Nitroaniline menunjukkan interaksi molekuler dan afinitas baik terhadap reseptor 1FIQ dengan nilai ?Gbind sebesar-6,6790 kkal/mol dan nilai Kd sebesar 8,8436 µM.; Hyperuricemia triggered by the overactivity of the xanthine oxidase enzyme, leads to severe cardiovascular diseases. Ethanol and aqueous extracts of bitter gourd (Momordica charantia L.) suppress uric acid synthesis, yet its molecular mechanism remains unexplored. This study identified the bioactive compounds from bitter gourd (NusaFarm) and evaluated their molecular interactions with the xanthine oxidase (PDB ID: 1FIQ) receptor in silico. Maceration using 96% ethanol followed by LC-HRMS analysis successfully annotated 132 compounds, predominantly carboxylic acids, with no major marker compounds detected. Minor marker compounds detected included coumarin and benzaldehyde. Bioavailability, ADME, toxicity prediction, and virtual screening (?Gbind and Kd) were evaluated and compared them against salicylic acid and allopurinol. Screening results revealed three non-toxic compounds; however, coumarin — which exhibited the best molecular interaction and affinity — was still visualized as a lead compound. 4-Nitroaniline demonstrated strong molecular interaction and good affinity toward the 1FIQ receptor, with a ?Gbind value of 6,6790 kcal/mol and a Kd value of 8,8436 µM. Thu, 01 Jan 2026 00:00:00 GMT http://repository.ipb.ac.id/handle/123456789/174105 2026-01-01T00:00:00Z http://repository.ipb.ac.id/handle/123456789/174038 Eksplorasi Gen Fungsional Bakteri Tanah pada Kebun Percobaan Kelapa Sawit Cikabayan IPB Dramaga secara Metagenomik Moureen, Annisa Julyantina Kebun Percobaan Kelapa Sawit Cikabayan adalah perkebunan monokultur tropis dengan kanopi terbuka, tanah asam, dan unsur hara esensial yang memengaruhi struktur dan fungsi komunitas bakteri tanah. Penelitian ini bertujuan mengeksplorasi keanekaragaman bakteri tanah dan gen fungsional melalui metagenomik. Metode: DNA tanah diekstraksi dengan FastDNA SPIN Kit for Soil, kualitas DNA dievaluasi melalui elektroforesis, fluorometri, dan spektrofotometri. Pustaka DNA disiapkan dengan Rapid Sequencing Kit DNA V14, disekuensing menggunakan PromethION 2 Solo (ONT), dan dianalisis dengan bioinformatika. Hasil menunjukkan data sekuensing berkualitas tinggi (Q19,3 dan Q16,5). Komunitas bakteri didominasi Streptomyces dan Bradyrhizobium. Tiga MAGs berkualitas sedang berpotensi sebagai spesies baru dengan gen fungsional terkait metabolisme asam amino, sistem transpor ABC, two-component system, resistensi beta-laktam, dan sintesis metabolit sekunder (terpen dan lantipeptida). Simpulan menunjukkan keragaman komunitas bakteri tanah dengan kapasitas metabolisme, adaptasi lingkungan dan aktivitas antropogenik, serta potensi biosintesis senyawa bioaktif sebagai bioinput untuk perkebunan berkelanjutan.; The Experimental Oil Palm Plantation Cikabayan is a tropical monoculture plantation characterized by an open canopy, acidic soils, and essential nutrients that influence the structure and function of soil bacterial communities. This study aimed to explore soil bacterial diversity and functional genes using metagenomics. Methods: Soil DNA was extracted using the FastDNA SPIN Kit for Soil, DNA quality was evaluated through electrophoresis, fluorometry, and spectrophotometry. DNA libraries were prepared using the Rapid Sequencing Kit DNA V14, sequenced on the PromethION 2 Solo (ONT), and analyzed using bioinformatics. The results showed high-quality sequencing data (Q19.3 and Q16.5). The bacterial community was dominated by Streptomyces and Bradyrhizobium. Three medium-quality MAGs were identified as potential novel species harboring functional genes associated with amino acid metabolism, ABC transport systems, two-component system, beta-lactam resistance, and secondary metabolite biosynthesis (terpenes and lanthipeptides). In conclusion, soil bacterial communities exhibit diversity with metabolic capacities, adaptability to environmental and anthropogenic activities, and biosynthetic potential for bioactive compounds as bioinputs for sustainable plantation systems. Thu, 01 Jan 2026 00:00:00 GMT http://repository.ipb.ac.id/handle/123456789/174038 2026-01-01T00:00:00Z http://repository.ipb.ac.id/handle/123456789/174009 Aktivitas Antioksidan dan Inhibitor Lipase Pankreas dari Ekstrak Etanol Buah Asam Jawa (Tamarindus indica L.) dan Daun Kelor (Moringa oleifera) ASTUTI, DEVI TULAK Obesitas sebagai masalah kesehatan global mendorong pengembangan potensi herbal dalam mekanismenya menginhibisi lipase pankreas untuk mengurangi penyerapan lemak. Senyawa bioaktif yang terkandung dalam buah asam jawa memiliki potensi sebagai inhibitor lipase pankreas dengan nilai persen inhibisi sebesar 68%. Sementara itu, daun kelor memiliki potensi yang serupa dengan nilai persen inhibisi sebesar 75,4%. Oleh karena itu, penelitian ini bertujuan menentukan kombinasi terbaik ekstrak buah asam jawa dan daun kelor berdasarkan kadar total fenolik, total flavonoid, aktivitas antioksidan, dan inhibisi lipase pankreas. Ekstraksi dilakukan menggunakan etanol 70% dengan metode ultrasonik, kemudian ekstrak dikombinasikan. Hasil penelitian ekstrak tunggal yang mendominasi hasil kadar fenolik total, flavonoid total, serta aktivitas antioksidan metode DPPH dan FRAP adalah ekstrak daun kelor dengan nilai berturut-turut 13,04 ± 0,60 mg GAE/g; 12,63 ± 0,00 mg QE/g; 5,610 ± 0,15 mg TE/g; 78,94 ± 1,15 mg TE/g. Akan tetapi, penghambatan lipase pankreas paling baik diperoleh pada ekstrak buah asam jawa (53,74%) dan kombinasi kedua ekstrak tidak menunjukkan efek sinergis yang signifikan.; Obesity as a global health problem ecourages the development of herbal agents that inhibit pancreatic lipase to reduce fat absorption. Bioactive compounds present in Tamarindus indica fruit have shown potential as pancreatic lipase inhibitors, with an inhibition percentage of 68%. Meanwhile, Moringa oleifera leaves exhibit similar potential, with an inhibition percentage of 75,4%. Therefore, this study aimed to determine the best combination of tamarind fruit and moringa leaf extracts based on total phenolic content, total flavonoid content, antioxidant capacity, and pancreatic lipase inhibition. Extraction was carried out using 70% ethanol assisted by ultrasonic waves, followed by the combination of the extracts. The result showed that the single extract dominating the TPC, TFC, antioxidant capacity (DPPH and FRAP methods) was the moringa leaf extract, with values respectively of 13,04 ± 0,60 mg GAE/g; 12,63 ± 0,00 mg QE/g; 5,610 ± 0,15 mg TE/g; 78,94 ± 1,15 mg TE/g, respectively. However, the highest pancreatic lipase inhibitory activity was observed in the tamarind fruit extract (53,74%), and the combination of both extracts did not show significant synergistic effects. Thu, 01 Jan 2026 00:00:00 GMT http://repository.ipb.ac.id/handle/123456789/174009 2026-01-01T00:00:00Z