Open AccessPemurnian Parsial dan Karakterisasi Kitinase Asal Jamur Entomopatogen Beauveria bassiana Isolat BB200109
Author(s) -
Yadi Suryadi,
Tri Puji Priyatno,
I Made Samudra,
Dwi Ningsih Susilowati,
Nuni Lawati,
Eman Kustaman
Publication year - 2016
Publication title -
jurnal agrobiogen/jurnal agrobiogen
Language(s) - English
Resource type - Journals
eISSN - 2549-1547
pISSN - 1907-1094
DOI - 10.21082/jbio.v9n2.2013.p77-84
Subject(s) - chitinase , bassiana , beauveria bassiana , chemistry , nuclear chemistry , food science , enzyme , biology , biochemistry , horticulture , biological pest control
Beauveria bassiana is one of the entomopathogenic fungus that produces chitinase when infecting its host. This study was aimed to purify, isolate and characterize chitinase of B. bassiana isolate BB200109. Pathogen identity was determined both morphologically and molecularly using ITS primer, whilst characterization was done at various conditions i.e. temperature, pH, metal ion and incubation time. Results showed that the BB200109 isolate belonged to B. bassiana. The isolate produced extracellular chitinase with chitinolytic index of 1.035. Partial purification of three saturated ammonium sulphate precipitation (10, 30, and 70%) showed maximum purity of 1.2 times, while dialysis could increase the purity of 1.9 times compared to that of crude enzyme extract. Characterization results showed that the chitinase isolated from B. bassiana isolate BB200109 had an optimum activity at pH 4, temperature 50oC, and optimum incubation time of 90 minutes. The effect of metal ions (60 mM) Mn2+ served as activator, while EDTA, K+, Mg2+, Cu2+, Fe2+, Zn2+, and Na+ acted as inhibitors. The chitinase demonstrated lower affinity to chitin substrate as indicated by high Km value of 0.266 mg/l and a Vmax of 0.067 mg/l sec. Based on SDS-PAGE, chitinase from B. bassiana isolate BB200109 had molecular weight of 60.25 kDa. The study implied the potency of B. bassiana isolate BB200109 as extracellular chitinase producer with its enzyme charateristics seems to be developed as an insect biocontrol agent.