ISOLASI ENZIM SELULASE dari TRICHODERMA B7 MENGGUNAKAN AMMONIUM SULFAT dan ETHANOL

Ika Fitriana, STP., Dr. Ir. M. Nur Cahyanto, M.Sc.

2013 | Tesis | S2 Ilmu dan Teknologi Pangan

Telah dilakukan penelitian isolasi enzim selulase dari Trichoderma B7 yang potensial menghasilkan enzim selulase, sehingga perlu dilakukan penelitian untuk mengisolasi enzim selulase hasil fermentasi menggunakan jamur Trichoderma B7 yang diinokulasi pada media Mendel, menggunakan sumber karbon CMC (Carboxy methyl cellulose sodium salt), fermentasi submerge selama 5 hari dan agitasi 120 rpm pada suhu 30ï‚°C. Setelah fermentasi dilakukan isolasi menggunakan ammonium sulfat dan ethanol pada kadar 50%, 60%, 70%, 80%, pH 4,5 dan suhu 0ï‚°C. Kemudian dilakukan pengujian kadar protein, aktivitas enzim, aktivitas spesifik terhadap enzim selulase setelah fermentasi dan setelah presipitasi kemudian dihitung nilai yield dan tingkat kemurniannya. Hasil penelitian menunjukkan enzim selulase setelah presipitasi menggunakan ammonium sulfat jenuh optimum aktivitas CMCase 0,189 mU/ml, kadar protein 19,797 mg/ml pada kadar presipitasi kadar 80%. Pada kadar presipitasi 50% optimum aktivitas specifik CMCase 0,034 mU/mg yield 89,6 %, dan tingkat kemurniannya 0,74. Enzim selulase setelah presipitasi menggunakan ethanol dengan hasil optimum pada kadar presipitasi 50% yaitu aktivitas CMCase 0,198 mU/ml, aktivitas specifik CMCase 0,083 mU/mg, yield 93,83% dan tingkat kemurniannya 2,52. Pada kadar presipitasi 70% optimum kadar protein sebesar 4,118 mg/ml.

Research has been conducted isolate of the fungus Trichoderma B7cellulase enzyme potentially produce cellulase enzymes mainly endoglukanase, so research must be done to isolate the cellulase enzyme fermentation produc from the fungus Trichoderma B7 were inoculated on media Mendel, carbon source from CMC (Carboxy methyl cellulose sodium salt), fermentation Submerge for 5 days and agitation of 120 rpm at a temperature of 30 ï‚° C. The enzymes from fermentation process isolate using ammonium sulfate at levels of 50% ethanol, 60%, 70%, 80%, pH 4.5 and a temperature of 0 ï‚° C. Then do the testing for protein content, enzyme activity, the specific activity of the enzyme cellulase after fermentation and after precipitation. Then calculated value of yields and purify of enzymes. Results of research the cellulase enzymes after isolation by using saturated ammonium sulfate precipitation optimum activity of CMCase 0,189 mU / ml, protein content 19,797 mg / ml in concentration levels of precipitation 80%. At optimum levels of precipitation 50% The specific activity of CMCase 0,034 mU / mg yield of 89,6%, and Purity 0,74. Cellulase enzymes after isolated by precipitation ethanol optimum at levels of precipitation 50%, 0,198 CMCase activity mU / ml, CMCase The specific activity of 0,083 mU / mg, 93,83% yield and purity 2.52. Optimum levels of precipitation 70% protein content 4,118 mg / ml.

Kata Kunci : Isolasi enzim, selulase, Trichodema B7, ammonium sulfat, ethanol