OPTIMASI DAN PEMEKATAN LIPASE Bacillus halodurans CM1 - Aisyah, Arina; Mangunwardoyo, Wibowo; Trismilah, Trismilah; Suhendar, Dadang (Department of Biology, Faculty of Science and Technology, Syarif Hidayatullah State Islami, 2017)
Abstrak Lipase diketahui memiliki peranan penting dalam bidang industri. Produksi lipase dapat dihasilkan oleh kapang, khamir, dan bakteri. Penelitian bertujuan untuk meningkatkan aktivitas lipase yang dihasilkan oleh Bacillus halodurans CM1. Aktivitas lipase dapat ditingkatkan dengan optimasi komposisi media, mutasi bakteri dengan radiasi gamma dan N-methyl-N’-nitro-N-nitrosoguanidine (NTG). Enzim yang dihasilkan dipekatkan dengan metode stirred-cell ultrafiltration (UF)-ammonium sulfat dan UF-Polyethylene glycol (PEG). Uji aktivitas dilakukan pada tujuh media yang berbeda untuk mendapatkan media produksi. Delapan variabel komposisi media dioptimasi dengan rancangan Plackett-Burman. Bakteri dimutasi dengan radiasi gamma dosis 0,1–0,4 kGy dan NTG 0,05–0,15 mg/mL dengan waktu inkubasi 1–3 jam. Hasil penelitian menunjukkan bahwa media produksi yang digunakan berdasarkan optimasi media dan komposisi media Plackett-Burman adalah media dasar Bora & Bora yang mengandung 0,5% palm oil (PO) dan 0,09% CaCl2. Aktivitas lipase optimal diproduksi oleh bakteri hasil mutasi dengan NTG 0,1 mg/mL yang diinkubasi selama 3 jam. Pemekatan enzim UF-ammonium sulfat dan UF-PEG mampu meningkatkan aktivitas enzim lipase sebesar 18,44%. Abstract Lipase is known to have an important role in the industrial field. Lipase can be produced by molds, yeasts, and bacteria. The research aimed to increase the activity of lipase produced by Bacillus halodurans CM1. Lipase activity can be improved by optimization of the composition of the media, the mutation of bacteria with gamma radiation and N-methyl-N'-nitro-N-nitrosoguanidine (NTG). The enzyme was concentrated by stirred-cell ultrafiltration method (UF)-ammonium sulfate and UF-Polyethylene glycol (PEG). The activity test was performed on seven different media to get production media. The eight variables of the media composition were optimized by Plackett-Burman design. The bacteria were subject to mutation by using 0.1–0.4 kGy dose of gamma radiation and 0.05–0.15 mg/mL NTG with incubation time for 1–3 hours. The results showed that the production media used based on optimization and composition of Plackett-Burman media was Bora Bora medium that containing 0.5% palm oil (PO) and 0.09% CaCl2. Optimum lipase activity was produced by the bacterium that mutated with 0.1 mg/mL NTG, incubated for 3 hours. The concentrated by UF-ammonium sulfate and UF-PEG could increase the lipase activity by 18.44%.