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Purification of Δ(5)-3-ketosteroid isomerase from Digitalis lanata.
MedLine Citation:
PMID:  25468533     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
The isomerization of 5-pregnene-3,20-dione into 4-pregnene-3,20-dione was investigated to shed further light on cardenolide biosynthesis and to characterize the enzymes involved in cardenolide formation. It was shown that the Δ(5)-3-ketosteroid isomerase of Digitalis lanata, which catalyzes this isomerization, is an individual enzyme and not, as previously thought, associated with Δ(5)-3β-hydroxysteroid dehydrogenase. The enzyme was purified by fractionated ammonium sulfate precipitation, hydrophobic interaction chromatography and gel filtration. The purification protocol resulted in a 68.1-fold enriched specific enzyme activity with a yield of 2.2%. After an additional chromatofocusing step the 3KSI activity appeared as a single protein band at 17kDa in SDS-PAGE. Plant 3KSI displayed similar properties to microbial 3-ketosteroid isomerases.
Authors:
Nadine Meitinger; Daniel Geiger; Thierry W Augusto; Rodrigo Maia de Pádua; Wolfgang Kreis
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2014-11-14
Journal Detail:
Title:  Phytochemistry     Volume:  109C     ISSN:  1873-3700     ISO Abbreviation:  Phytochemistry     Publication Date:  2014 Nov 
Date Detail:
Created Date:  2014-12-3     Completed Date:  -     Revised Date:  2014-12-4    
Medline Journal Info:
Nlm Unique ID:  0151434     Medline TA:  Phytochemistry     Country:  -    
Other Details:
Languages:  ENG     Pagination:  6-13     Citation Subset:  -    
Copyright Information:
Copyright © 2014 Elsevier Ltd. All rights reserved.
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