PRODUCTION AND CHARACTERIZATION OF CELLULOSE ENZYME ISOLATED FROM LOCAL ISOLATE OF CYTOPHAGA BACTERIA

Abstract

Abstract: The aim of the research was to produce Cellulase via the use of two isolates of Cytophaga bacteria. The first isolate, given the local code CR1, was taken from AL-gazeera region soil in Ramadi grown with okra. The second, given the local code CR2, was taken from animal waste. Wheat barn and corn wastes (grimes) were used as only carbon source in the culture media where the two isolates have been cultivated. However these wastes were available in big amounts in this environment. CR2 showed a great ability to produce Cellulase in the liquid medium culture .The best production was at pH 7.5 and 35c temperature using wheat barn of 1.5% cellulose concentration. When incubated in an incubator shaker of 125 R.p.m and a bacterial density of 1ml/100ml medium. After 5 days of incubation, the enzymatic activity was 5.976 unit/ml. The results have shown that adding calcium and magnesium ions as sulfates of 0.5g/100ml,concentration gave higher production of the enzyme. The enzyme activity was 6.121unit/ml and 6.212unit/ml,respectively. The enzyme produced from the local isolate CR2 was purified following purification procedure that included precipitation using ammonium sulfates of 90% saturation and ion exchange chromatography via replacing the column DEAE-Sepharose.This was followed by gel filtration column of Superdex-200.It was possible 64.11% of the enzyme with 4.109 times of purification. The results of enzyme description showed that its molecular weight was about 32000 Dalton using gel filtration chromatography of a Superdex-200 column, and that the optimum pH of the enzyme activity was 6.0 .The optimum temperature of the enzyme activity was 50c .The results indicated, also, that the enzyme gave the highest enzymatic activity after 60 minutes of incubation at a pH6.0 and 50c temperature.