Document Detail


Affinity precipitation of Aspergillus niger pectinase by microwave-treated alginate.
MedLine Citation:
PMID:  14680967     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Affinity precipitation is a simple, single plate separation process in which the complex of a smart macroaffinity ligand with the target protein (from a crude broth) can be selectively precipitated by application of a suitable stimulus. Alginate is a copolymer of guluronic acid and mannuronic acid residues and precipitates with Ca(2+) ions. It was found to bind to pectinase present in a commercial preparation of Aspergillus niger, Pectinex Ultra-SPL. Microwave pretreatment of alginate at 75 degrees C was found to enhance the selectivity of the affinity precipitation. Using microwave-treated alginate, 83% of the enzyme activity with 20-fold purification could be recovered. SDS-PAGE upon silver staining confirmed the enhanced selectivity of affinity precipitation when microwave-treated alginate was used.
Authors:
Kalyani Mondal; Payal Mehta; Munishwar N Gupta
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Protein expression and purification     Volume:  33     ISSN:  1046-5928     ISO Abbreviation:  Protein Expr. Purif.     Publication Date:  2004 Jan 
Date Detail:
Created Date:  2003-12-18     Completed Date:  2004-04-27     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  9101496     Medline TA:  Protein Expr Purif     Country:  United States    
Other Details:
Languages:  eng     Pagination:  104-9     Citation Subset:  IM    
Affiliation:
Department of Chemistry, Indian Institute of Technology, Delhi, Hauz Khas, New Delhi 110016, India.
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MeSH Terms
Descriptor/Qualifier:
Alginates / chemistry,  metabolism*
Aspergillus niger / enzymology*
Calcium Chloride / chemistry
Chemical Precipitation
Electrophoresis, Polyacrylamide Gel
Glucuronic Acid / chemistry,  metabolism*
Hexuronic Acids / chemistry,  metabolism*
Microwaves
Polygalacturonase / isolation & purification*,  metabolism
Protein Binding
Spectrophotometry, Ultraviolet
Temperature
Chemical
Reg. No./Substance:
0/Alginates; 0/Hexuronic Acids; 10043-52-4/Calcium Chloride; 576-37-4/Glucuronic Acid; 9005-32-7/alginic acid; EC 3.2.1.15/Polygalacturonase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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