PEMURNIAN PARSIAL EKSTRAK KASAR SELULASE BACILLUS CIRCULANS DENGAN METODA PENGENDAPAN ASETON
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Tujuan penelitian ini adalah untuk mengetahui tingkat kemurnian hasil pengendapan ekstrak kasar selulase Bacillus circulans pada berbagai kejenuhan aseton; pengaruh waktu pengendapan terhadap tingkat kemurnian hasil pengendapan pada kejenuhan aseton optimum; dan stabilitas waktu penyimpanan fraksi optimum pada suhu 0°C selama 0,1,4 dan 7 hari.Aktivitas selulase diukur berdasarkan kenaikan kadar gula pereduksi yang dihasilkan pada substrat kertas Whatman no 1. Penentuan kadar gula pereduksi menggunakan metode Somogy-Nelson. Kadar protein ditentukan dengan metode Lowry. Tingkat kemurnian enzim merupakan perbandingan aktivitas spesifik enzim setelah pemurnian dengan sebelum pemurnian.Tingkat kemurnian hasil pengendapan ekstrak kasarBacillus circulans mengalami peningkatan pada kejenuhan aseton ( 33 %), (41,1 %) dan (50 %) kemudian menurun pada kejenuhan (60 %) dan (67 %). Fraksi yang aktivitas spesifik tertinggi adalah pada tingkat kejenuhan aseton 50 %, dengan tingkat kemurnian rata-rata 63,61 ± 0,141 kali ekstrak kasar dan perolehan protein kembali rata-rata 76,85 ± 7,99 %. Waktu pengendapan tidak berpengaruh terhadap tingkat kemurnian hasil pengendapan ekstrak kasar selulase Bacillus circulans pada kejenuhan aseton optimum.Selulase hasil pengendapan dengan aseton memiliki stabilitas yang kurang baik pada suhu ± 0°C karena menurun sebesar 41,1 % pada hari pertama dan 88,2 % pada hari ketujuh setelah penyimpanan.
Kata kunci: selulase, Bacillus circulans, pengendapan, aseton.
Abstract
The aims of this study were to determine the purity level of precipitations of cellulases of Bacillus circulans at various saturation of acetone; the effect of precipitation time on the purity level of the precipitations at optimum acetone saturation; and the stability of the storage time of the optimum fraction at 0 ° C for 0,1,4 and 7 days. The activity of cellulase was measured based on the increased of reducing sugar levels produced from hydrolysed of Whatman paper as substrate. The reducing sugar levels were determined with the Somogy-Nelson method. Protein levels are determined with the Lowry method. The purity level of the enzyme is a comparison of specific enzyme activity after and before purification. The purity level of precipitations of cellulose of Bacillus circulans has increased in saturation of acetone (33%), (41.1%) and (50%) then decreases in saturation (60%) and (67%). The highest cellulase specific activity fraction was at 50% acetone saturation level, with an average purity level of 63.61 ± 0.141 times crude extract and yield protein recovery an average of 76.85 ± 7.99%. The precipitation time did not affect the purity level of the precipitations at optimum acetone saturation. The precipitations had low storage stability at ± 0 oC because its cellulase activity decreased by 41.1% on the first day and 88.2 % on the seventh day after storage
The keywords: cellulose, Bacillus circulans, precipitation, aceton
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