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Isolation, Purification and Partial Characterization of DDMS- Dehydrochlorinase from Pseudomonas Dimunita
Hazel, Preethi J. (2009) Isolation, Purification and Partial Characterization of DDMS- Dehydrochlorinase from Pseudomonas Dimunita. [Student Project Report]
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Abstract
This Dissertation / Report is the outcome of investigation carried out by the creator(s) / author(s) at the department/division of Central Food Technological Research Institute (CFTRI), Mysore mentioned below in this page.
| Item Type: | Student Project Report |
|---|---|
| Additional Information: | Ten bacterial isolates of the DDT-degrading microbial consortium were screened for their ability to dechlorinate DDMS [1-chloro-2, 2-bis (p-chlorophenyl) ethane], a metabolite in the DDT degrading pathway to DDNU [unsym-bis(4'-Chlorophenyl)-ethylene]. Based on the dechlorinase activity, a pure isolate Pseudomonas dimunita was selected for further studies. The enzyme studies were performed with cell free extracts. The enzyme was purified by size exclusion chromatography. On SDS-PAGE the purified enzyme showed a single band. The molecular weight as per SDS-PAGE was found to be around 45kDa.The purity of the enzyme was checked by reverse phase HPLC using C18 column. The dehydrochlorinase activity was maximum at 750 μgml−1 substrate concentration, pH 7 & temperature 20ºC. DDMS-dehydrochlorinase was found to be present in two isoforms & therefore exhibited two peaks. It had a narrow range of specificity towards other halogenated substrates such as Endrin; Dieldrin; α, ß, γ & δ-HCH; 2, 4, 6-Trichlorophenol & Chlorobenzene. The enzyme appeared to be a glycoprotein, peak 1 & peak 2 was found to have 76% & 31% of glycoprotein respectively. The spectroscopic studies such as UV-Visible, fluorescence and FTIR were also done. |
| Uncontrolled Keywords: | Microbial degradation DDT dechlorinate DDMS Pseudomonas dimunita |
| Subjects: | 600 Technology > 08 Food technology > 16 Nutritive value > 05 Enzymes 500 Natural Sciences and Mathematics > 07 Life Sciences > 04 Microbiology |
| Divisions: | Fermentation Technology and Bioengineering |
| Depositing User: | Food Sci. & Technol. Information Services |
| Date Deposited: | 16 Sep 2009 11:26 |
| Last Modified: | 28 Dec 2011 10:11 |
| URI: | http://ir.cftri.res.in/id/eprint/9156 |
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