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STATISTICAL OPTIMIZATION OF SYNTHETIC AZO DYE (ORANGE II) DEGRADATION BY AZOREDUCTASE FROM Pseudomonas oleovorans PAMD_1.
MedLine Citation:
PMID:  23768111     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Pseudomonas oleovorans PAMD_1 produced an intracellular azoreductase as the more prominent enzyme that reduces the azo bridge during the azo dye decolorization process. In order to optimize the expression of azoreductase, statistically based experiments were applied. Eleven significant factors were screened on decolorization activity using Plackett-Burman design. Dye, NADH, glucose, and peptone were identified as having highest positive influence on the decolorization activity. Central composite design of response surface methodology was employed for the concerted effect of these four factors on decolorization activity. This method showed that the optimum medium containing dye (200 mg L(-1)), NADH (1.14 mM), glucose (2.07 g L(-1)), and peptone (6.44 g L(-1)) for the decolorization of Orange II up to 87% in 48 hr. The applied methodology was validated through the adequacy and accuracy of the overall experiments, and the results proved that the applied methods were most effective. Further, the enzyme was purified ninefold with 16% yield by anion-exchange chromatography and a specific activity of 26 U mg(-1). The purified enzyme with a molecular mass of 29,000 Da gave a single band on sodium dodecyl sulfate (SDS) gel, and the degradation products sulfanilic acid and 1-amino-2-napthol of Orange II by azoreductase were analyzed by using an ultraviolet-visible (UV-Vis) spectrophotometer and hish-performance liquid chromatography (HPLC).
Authors:
V Aranganathan; A M Kanimozhi; T Palvannan
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Preparative biochemistry & biotechnology     Volume:  43     ISSN:  1532-2297     ISO Abbreviation:  Prep. Biochem. Biotechnol.     Publication Date:  2013 Oct 
Date Detail:
Created Date:  2013-06-17     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9607037     Medline TA:  Prep Biochem Biotechnol     Country:  England    
Other Details:
Languages:  eng     Pagination:  649-67     Citation Subset:  IM    
Affiliation:
a Department of Biochemistry , Center for Postgraduate Studies, Jain University , Bangalore , Karnataka , India.
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