Effect of Denaturants on the Structure and Activity of DDT Dehydrochlorinase.

Latha, R. and Mandappa, I. M. and Thakur, M. S. and Manonmani, H. K. (2011) Effect of Denaturants on the Structure and Activity of DDT Dehydrochlorinase. Asian Journal of Microbiology, Biotechnology and Environmental Sciences, 13 (1). pp. 51-57.

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Abstract

Urea, sodium dodecyl sulfate (50s) and guanidine hydrochloride (GdmCl) are often used as excellent denaturing or "unfolding" agents. We studied the effect of these denaturing agents on the structural changes of DDT- enzyme of Psuedomonas pulido. TS. There was a progressive loss in catalytic activity of DDT- dehydrochlorinase with increasing concentrations of denaturants, namely urea, 50s and GdmQ. At 10 M urea, 5 % 50s and 1 M GdmCl, the extent of loss in enzyme activity was 98, 78 and 100% respectively. The emission spectrum of urea denatured enzyme did not show shift, but that of 50s and GdmCl treated enzyme showed vel)' marginal shift. The secondary structure analysis of the enzyme by CD spectrum suggested a predominance of l1-structure in the untreated enzyme. Urea denatured enzyme revealed complete loss of «-helix and there was substantial reduction in 11 sheets. SDS treated enzyme showed increase in a-helix, turns and random structure. However, there was a marginal loss in 11 sheets. GdmCl treated enzyme showed complete absence of a-helix compared to the control enzyme. The l1-sheets were also reduced and the number of turns increased. The effects of GdmCI were almost similar to that of urea as observed in CO spectrum. The urea polyacrylamide gel showed only one band whereas 505- polyacrylamide gel showed closely moving two bands.

Item Type:	Article
Uncontrolled Keywords:	DDT- dehydrochlorinase, Denaturants, Urea, Sodium dodecyl sulphate, Guanidine hydrochloride, Enzyme activity
Subjects:	600 Technology > 05 Chemical engineering > 03 Enzyme Biotechnology and Engineering
Depositing User:	Food Sci. & Technol. Information Services
Date Deposited:	28 Jun 2017 06:04
Last Modified:	28 Jun 2017 06:04
URI:	http://ir.cftri.res.in/id/eprint/12688

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