Isolation and Purifi cation of Chitinase Bacillus sp. D2 Isolated from Potato Rhizosfer
Sebastian Margino(1*), Chatarina Behar(2), Widya Asmara(3)
(1) 
(2) 
(3) 
(*) Corresponding Author
Abstract
Potato Cyst Nematodes (Globodera rostochiensis) is one of the important potato’s pests and caused economic
looses up to 70% in the several centrals of potato plantations in Indonesia. Potato Cyst Nematodes (PCN) shell
component of egg shell containing chitin (inner layer) and vitelline/protein (outer layer), so the purpose of
research was to fi nd out of chitin degrading bacteria for controlling of egg’s PCN by cutting of their life cycle.
The results showed that Bacillus sp. D2 isolated from potato rhizosphere could produce extra cellular chitinase
in the medium containing of 0.20% colloidal chitin and fermented for 72 hours. Result of chitinase purifi cation
using ammonium sulphate precipitation and DEAE-Cellulose ion-exchange chromatography showed a specifi c
activity 2691,052 U/mg and analyzing using SDS-PAGE 12.5% resulted in molecular weight 30 kDa. The
apparent Km and Vmax of chitinase towards colloidal chitin were 2 mg/ml and 2.2 μg/h, respectively.
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PDFDOI: https://doi.org/10.22146/ijbiotech.7851
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