Paper Title
The Effectiveness of the Enzyme Chitinase Isolated and the Purification Process with the Stages 3 Species Vibrio sp. As anti-fungus fusarium oxysporium

The process of chitin degradation by Vibrio has adverse effects on marine ecosystems, as bacteria cause a number of diseases, especially in animals containing chitin as the components of the exoskeleton. Chitin degradation to N-acetyl-D-glucosamine has great benefits in agriculture as an anti-fungus. The aims of this research is to know the activity and purification of enzyme using precipitation method of ammonium sulfate and Sephadex G-75 chromatography column and to study chitinase role of Vibrio alginolyticus, V.fluvialis and V.harveyi as anti-fungi for Fusarium fungi and 3 species of Trichoderma as in vitro. This research consists of 2 stages of research. The first stage of the study was done 2 purification of rough enzymes, namely: a) precipitation method of ammonium sulfate with variation of treatment that is 0-30%; 30-45% and 45-60%, followed by dialysis and Sephadex 75G chromatographic column method (15 elution time / minute x 3 Vibrio sp.); b) Specific enzyme by dot blotting method at 0.02 dilution; 0.2 and 2 mL / mol and western blotting using monoclonal antibody chitinase. The second stage of the study was done application of chitinase as an anti-fungi for plant disease control by Fusarium oxysporium fungi and 3 species of Trichoderma (T.harzianum, T.koningii and T.viridae) in vitro. The results showed that the enzyme purification method with the saturated fraction of saturated ammonium sulfate and chromatographic column had the highest different activity for 3 Vibrio species. Vibrio alginolyticus at 0-30% precipitation fraction and elution time of 11th min, while V.fluvialis and V.harveyi at 30-45% deposition fraction and elution time of 4th and 6th min. The results of chitinase spesificity testing by western blotting method yielded 2 electrophoresis bands with molecular weight of 60-65 kDa and 57-59 kDa. The isolated chitinase of 3 Vibrio species can be applied to inhibit the growth of Fusarium oxysporium pathogens. Key word - Chitinase, Isolated, Purification, Vibrio sp., Fusarium oxysporium, Anti-fungi