ISOLATION AND CHARACTERIZATION OF HALOACID DEHALOGENASE FROM PSEUDOMONAS AERUGINOSA LOCAL STRAIN

• Main Author: Ulma'awi, Uun
• Format: Final Project
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/34542
• Institution: Institut Teknologi Bandung

Halogenated compound is one of environmental pollutants. This compound widely used as intermediate compounds in fertilizer, pharmaceutical, and cosmetics. Bioremediation is one important method used to reduce pollutants. Pseudomonas aeruginosa is a potensial organism to be used as a bioremediator as it is known to produce dehalogenase. This research studied haloacid dehalogenase from Pseudomonas aeruginosa local strain. Dehalogenase assay is performed by colorimetric method using monochloroacetic acid as substrate. The result indicated that intracellular dehalogenase from this bacterium has a specific activity of 0,0398 U/mg protein (U = µmol chloride released during reaction per mL per minute). Fractionation using 60-80% ammonium sulphate to the intracellular crude extract enzyme yielded highest dehalogenase specific activity which is 0,0635 U/mg protein. Dehalogenase in this fraction have optimum pH at 8 and optimum temperature at 37 oC. Kinetics measurements at various substrate concentration gives Km 93,2281 mM and Vmax 0,49036 U/mg. Compared with dehalogenase from Pseudomonas putida strain AJ1/23 (Km = 27,5 mM), Pseudomonas aeruginosa strain local has low affinity to MCA (Km = 93,2281 mM).