Isolasi dan karakterisasi Enzim Protease dari Aspergillus oryzaaae 6004.

Mediana , Vanny (2000) Isolasi dan karakterisasi Enzim Protease dari Aspergillus oryzaaae 6004. Undergraduate thesis, FMIPA UNDIP.

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Abstract

Enzim protease adalah enzim yang dapat menghidrolisis ikatan peptida dalam protein. Telah dilakukan isolasi dan karakterisasi enzim protease dari jamur .4spergillus oryzae 6004. Jamur Aspergillus oryzae ditumbuhkan pada media PDA (Potato Dextrose Agar) kemudian difennentasikan pada media campuran dedak dan tepung kedelai. isolasi dilakukan dengan metode ekstraksi. Hasil isolasi difraksinasi bertingkat menggunakan ammonium sulfat, yaitu F 1 (0-10 %), F2 (10-30 %), F3 (30-50 %), F4 (50-70 %) dan F5 (70-100 %), dilanjutkan dialisis dalam bufer fosfat. Fraksi yang didapat diuji dengan substrat kasein, menggunakan Spektrofotometer UV-Vis pada panjang gelombang maksimum tirosin dan hasilnya diekstrapolasikan terhadap kurva standar tirosin, untuk mendapatkan nilai aktivitas enzim protease. Satu unit aktivitas didefinisikan sebagai aktivitas enzim yang menyebabkan terbentuknya 1 jamol tirosin per satuan waktu inkubasi pada kondisi optimum. Selain itu ditentukan aktivitas spesifik, yaitu unit aktivitas tiap milligram protein. Kadar protein ditentukan dengan metode Lowry. Karakterisasi yang dilakukan meliputi penentuan temperatur, pH, dan waktu inkubasi optimum. Hasil penelitian menunjukkan bahwa fermentasi optimum pada 72 jam dan fraksi 2 (F2 dengan tingkat kejenuhan 10-30 %) memiliki tingkat kemurnian yang tertinggi dengan aktivitas spesifik 189,0818 Unit/mg. Berdasarkan penentuan sifat karakteristiknya, enzim protease dari Aspergillus oryzae 6004 ini bekerja optimal pada temperatur 33 °C, pH 7,5, dan waktu inkubasi selama 15 menit. Protease enzyme is an enzyme which able to hydrolize peptide bonding in protein, Isolation and characterization of protease enzyme have been done from soy sauce mould (Aspergillus oryzae 6004). Aspergillus oryzae was planted in PDA (Potato Dextrose Agar) medium, and fermented in mixture media of dedak (mixture of rice and bran) and soy flour. Isoaltion has been carried out by extraction method. It has been done phased fractination using ammonium sulphate, that are F1 (0-10 %), F2 (10-30 %), F3 (30-50 %), F4 (50-70 %), and F5 (70-100 %), continued to dialysis in phospate buffer. The obtained fraction analyzed using casein substrate by UV-Vis Spectrophotometer in tyrosin maximum wavelength and its result extrapolized to tyrosin standart curve to obtain protease enzyme activity value. Beside, it was determined the spesific activity, activity unit per milligram of protein. Protein degree was determined by Lowry method. The characterization had done are determination of optimum temperature, pH, and incubation time. The research result showed that optimum fermentation is at 72 hours and fraction 2 (F2 by unsaturated degree 10-30 %) have highest pure degree with spesific activity 189.0818 Unit/mg. Based on characterization determination, this enzyme active optimally in temperature 33 °C, pH 7.5, and incubation time 15 minutes.

Item Type:Thesis (Undergraduate)
Subjects:Q Science > QD Chemistry
Divisions:Faculty of Science and Mathematics > Department of Chemistry
ID Code:30797
Deposited By:Mr UPT Perpus 1
Deposited On:07 Nov 2011 11:03
Last Modified:07 Nov 2011 11:03

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