Malaysian Journal of Microbiology  2018;14(3):244-253

Purification and characterization of dehalogenase from Bacillus cereus SN1 isolated from cow dung

Siti Nurul Fasehah Ismail 1 ; Mohamed Faraj Edbeib 2 ; Wan Mohd Khairul ikhsan Wan Seman 3 ; Mahzan Md. Tab 3 ; Farahayu Khairuddin 3 ; Amin Retnoningsih 4 ; Roswanira Abdul Wahab 1 ; FahrulHuyop 4

Affiliations

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Keywords

Cow dung; dehalogenase; 2,2-dichloropropionic acid; Bacilluscereus SN1; bioremediation

Country

Malaysia

Language

English

Abstract

Aims:This study was aimed to characterize a dehalogenase derived from Bacillus cereus SN1 isolated from cow dung.

Methodology and results:Cell-free extract of Bacillus cereus SN1 was purified using ion exchange and gel filtration chromatography. Fraction B2 of gel filtration gave the highest enzyme specific activity (0.155 μmol CI¯/min/mg). The results of SDS-PAGE showed the enzyme was 25 kDa in size. The enzyme reached its optimum activity at 30 °C at pH 6, and was inhibited by Mercury(II) sulfate (HgSO4). The Km and kcat values were 0.2 mM and 1.22/sec, respectively. The partial dehalogenase gene sequence was amplified using Group I dehalogenase primers. The amplified gene sequence was designated as DehSN1.

Conclusion, significance and impact of study:Dehalogenase from Bacillus cereus strain SN1 revealed new characteristics of dehalogenase protein. The findings indicated that the DehSN1 dehalogenase is a promising candidate for further studies as a bioremediation agent for agricultural applications.