Document Detail


Characterization of a novel resistance related deoxycytidine deaminase from Brassica oleracea var. capitata.
MedLine Citation:
PMID:  24475736     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Brassica oleracea deoxycytidine deaminase (BoDCD), a deoxycytidine deaminase (DCD, EC 3.5.4.14) enzyme, is known to play an important role in the Trichoderma harzianum ETS 323 mediated resistance mechanism in young leaves of Brassica oleracea var. capitata during Rhizoctonia solani infection. BoDCD potentially neutralizes cytotoxic products of host lipoxygenase activity and thereby BoDCD restricts the hypersensitivity related program cell death induced in plants during the initial stages of infection. To determine the biochemical characteristics and to partially elucidate the designated functional properties of BoDCD, the enzyme was cloned into an E. coli expression system and their potentiality to neutralize the toxic analogs of 2'-deoxycytidine was examined. BoDCD transformants of E. coli cells were found to be resistant to 2'-deoxycytidine analogs in all the concentrations tested. The BoDCD enzyme was also over-expressed as a histidine-tagged protein and purified using Nickel chelating affinity chromatography. The molecular weight of BoDCD was determined to be 20.8 kDa as visualized by SDS-PAGE. The substrate specificity and other kinetic properties show that BoDCD is more active in neutralizing cytotoxic cytosine β-D-arabinofuranoside than in deaminating 2'-deoxycytinde to 2'-deoxyuridine in nucleic acids or in metabolizing cytidine to uridine. The optimal temperature and pH of the enzyme were 27 °C and 7.5. The Km and Vmax values of BoDCD were respectively, 91.3 µM and 1.475 mM for its natural substrate 2'deoxycytidine; 63 µM and 2.072 mM for cytosine β-D-arabinofuranoside. The phenomenon of neutralization of cytotoxic dC analogs by BoDCD is discussed in detail based on the enzyme biochemical properties.
Authors:
Shibu Marthandam Asokan; Hong-Shin Lin; Chaur-Tsuen Lo; Shu-Ying Liu; Kou-Cheng Peng
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2014-1-29
Journal Detail:
Title:  Journal of agricultural and food chemistry     Volume:  -     ISSN:  1520-5118     ISO Abbreviation:  J. Agric. Food Chem.     Publication Date:  2014 Jan 
Date Detail:
Created Date:  2014-1-30     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0374755     Medline TA:  J Agric Food Chem     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
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