Manuhara, Yosephine Sri Wulan and Puspaningsih, Ni Nyoman Tri and Wahyuningsih, Sri Pudji Astuti (2009) Purification of β-1,3-Endoglucanase from Cabbage (Brassicaoleracea cv. capitata L.) by Ion Exchange Chromatography. In: Second International Conference and Workshops on Basic and Applied Sciences and Regional Annual Fundamental Science Seminar 2009 volume iv, 2-4 June 2009, Johor Bahru, Malaysia.

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Abstract

In this research, there was purified of β-1,3- endoglucanase from cabbage, having the aim to calculate purification level of enzyme by ion exchange chromatography methods. That process was started by producing enzyme isolated from Gloria osena hybrid of cabbage,then precipitate it using ammonium sulphate with concentration of saturated 40%. The sediment of enzyme was diluted by phosphate citrate then purified by dialysis in order to separate enzyme from other proteins and ammonium sulphate. The next, hydrophobic interaction chromatography eluted by ammonium suphate concentration of saturated 40% in Tris HCl pH 7 (high concentration to low concentration gradient) with Butyl-topearl 650M in ethanol as matrix was done to purify enzyme based on hydrophobic group interaction of protein and absorbent. After that, there was purifying enzyme by ion exchange chromatography in order to separate enzyme based on it’s ion. Enzyme was eluted by NaCl (0- 0.5) M in Tris HCl (low concentration to high concentration gradient) with DEAE-toyopearl 650 M in ethanol as the matrix. The best result of this ion exchange chromatography on first fraction pH 7 had purification level 3.534,1 of initial extract.

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