Purification and Characterization of Linamarse from Gadung (Dioscorea hispida Dennst) for Gadung Slurry Detoxification

Harijono, Harijono
N., Siwi
Sutrisno, Aji

Jurnal Teknologi Pertanian
Vol 12, No 2 (2011)

The aim of this research was to find out purification of enzyme linamarase fromDioscorea hispida Dennts. Enzyme purification was carried out by amonium sulphatefractionation and Sephadex G-75 gel filtration coloumn chromatography. Protein purifyfactor achieved was 10.5 times, protein yield was 33.3 % of the crude cell extract proteinand specific activity was 2.93 U/mg. The enzyme purity of each purification stage wasmonitored by SDS-PAGE and resulted one protein band. The optimum temperature andpH of enzyme reaction was 57°C and pH 6.5 with 90 minutes incubation. Using crudelinamarin as substrat, linamarase has Km 0.1564 μmol ml and Vmax 0.2796μmol/minute. Pure linamarase was added in Dioscorea slurry to increase cyaniderelease. Keywords: purification, characterization, linamarase, detoxification, Dioscorea hispidaDennts.