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

• Main Authors: Ismail, Siti Nurul Fasehah, Edbeib, Mohamed Faraj, Wan Seman, Wan Mohd. Khairul ikhsan, Md. Tab, Mahzan, Khairuddin, Farahayu, Retnoningsih, Amin, Abdul Wahab, Roswanira, Huyop, Fahrul
• Format: Article
• Language: English
• Published: Universiti Sains Malaysia 2018
• Subjects: QH301 Biology; TP Chemical technology
• Online Access: http://eprints.utm.my/id/eprint/85695/1/SitiNurulFasehah2018_PurificationandCharacterizationofDehalogenasefromBacillusCereus.pdf; http://eprints.utm.my/id/eprint/85695/; http://dx.doi.org/10.21161/mjm.113517

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.