Abstract
This research aims to produce large amounts of lipase and have high activity so that it can be utilized in the food industry. Method of Research used a thermostable enzyme derived from compost AL96 microorganisms. Qualitative tests are carried out using Thermus media. Extracellular lipase enzyme expressed by AL96 culture at 70 0 C for 17 hours was isolated as much as 1000 mL and the crude extract obtained was deposited by fractionation of ammonium sulfate. After obtaining a fraction of 0-30%; 30 -50% fraction; 50-70% fraction; and 7-90% fraction was then tested for lipase enzyme activity using spectrophotometric techniques and testing of protein content determined by the Bradford method. Results Of research obtaining partial purification using ammonium sulfate into crude enzyme extract from AL96 isolate with a fraction of 50-70% resulted in the highest specific activity of protein 0.018 U / mg. Further analysis of lipase in the 30-50% fraction has the optimum temperature at 65 0 C and the fraction of 50-70% has the optimum temperature at 75 0 C. Characterization of the optimum pH for 50-70% fraction and 30-50% fraction showed that both fractions had optimum pH 10.
Highlights
so that it can be utilized in the food industry
Method of Research used a thermostable enzyme derived from compost AL96 microorganisms
Qualitative tests are carried out using Thermus media
Summary
Of research obtaining partial purification using ammonium sulfate into crude enzyme extract from AL96 isolate with a fraction of 50-70% resulted in the highest specific activity of protein 0.018 U / mg. Produksi enzim dimulai dengan membuat media stater sebanyak 10 mL, kemudian satu koloni bakteri dari media padat dipindahkan kedalam media stater, inkubasi dilakukan dalam inkubator kocok pada suhu 55 oC selama 17 jam dengan kecepatan 150 rpm. Fraksinasi kemudian dilakukan pada ruang dingin, amonium sulfat ditambahkan sedikit demi sedikit ke dalam ekstrak kasar enzim sambil diaduk dengan magnetic stirrer campuran didiamkan selama ± 3 jam untuk menyempurnakan pengendapan protein, kemudian disentrifugasi dengan kecepatan 10000xg selama 20 menit pada temperatur 4 oC. Sebanyak 900 μL Larutan tersebut kemudian ditambahkan denga 300 μL larutan enzim dan diinkubasi pada suhu 55 oC dengan menggunakan inkubator selama 15 menit. Penentuan pengaruh pH terhadap aktivitas lipase dilakukan pada temperatur 55oC menggunakan 50 mM bufer dengan rentang pH dari 6-12. Campuran reaksi diinkubasi pada temperatur tertentu dalam penangas air, dan aktivitas lipase diukur mengikuti prosedur yang dikemukakan oleh Lee dkk. Campuran reaksi diinkubasi pada temperatur tertentu dalam penangas air, dan aktivitas lipase diukur mengikuti prosedur yang dikemukakan oleh Lee dkk. (1999)
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