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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.com/id/eprint/9156

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