ISOLATION AND CHARACTERIZATION OF HALOALKANE DEHALOGENASE FROM A LOCAL STRAIN OF PSEUDOMONAS AERUGINOSA
Organohalide compound is widely used in pesticide indutstry, fuel additive, intermediet for chemical synthesis and solvent. One of the most abundant organohalide compound in the enviroment is 1,2-dichloethane (DCE). Organohalide is persistent and toxic to the enviroment. Therefore, organoh...
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| Format: | Final Project |
| Language: | Indonesia |
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| Online Access: | https://digilib.itb.ac.id/gdl/view/34445 |
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| Institution: | Institut Teknologi Bandung |
| Language: | Indonesia |
| Summary: | Organohalide compound is widely used in pesticide indutstry, fuel additive, intermediet for chemical synthesis and solvent. One of the most abundant organohalide compound in the enviroment is 1,2-dichloethane (DCE). Organohalide is persistent and toxic to the enviroment. Therefore, organohalide is a pollutant, hazard to the enviroments as it’s difficult to be degraded and suspected as a carsinogen to human. Several bacteria has been identified as able to degrade DCE, such as bacteria from genus Pseudomonas, Bradyrhizobium, and Xantobacter. These bacteria produce haloalkane dehalogenase which catalyze the breakdown of covalent bond between carbon and halide in organohalide compound. In DCE dehalogenase catalysis produces 2-chloroethanol, halide ion, and proton. In this research, isolation and characterization of haloalkana dehalogenase from Pseudomonas aeruginosa local strain were carried out. Pseudomonas aeruginosa was identified by PCR 16S ribosomal DNA using Unibi (5’-GGT TAC (GC) TTG TTA CGA CTT-3) as a forward primer and BactF-1 (5’-AGA GTT TGA TCA CTG GCT CAG-3’) as a reverse primer. It was observed that Pseudomonas aeruginosa able to grow in Luria Bertani and minimal medium containing
1?10mM DCE. Haloalkane dehalogenase is found to be produced intraselularly with specific activity of 8,809 unit/mg protein (unit = µmol Cl- / minute). Haloalkane dehalogenase fractination resulted in increasing specific activity to 32,108 unit/mg protein which is 3,5 times higher compare to crude extract. This haloalkane dehalogenase has an optimum pH 8 and optimum temperature 30 0C. Pseudomonas aeruginosa local strain has a greater potential for bioremediation than Pseudomonas aeruginosa OK1, because it’s specific activity is higher and it’s able to grow faster than OK1.
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