Document Detail


Rational improvement of cell-free protein synthesis.
MedLine Citation:
PMID:  20603235     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Experimental design principles were applied on cell-free protein synthesis to optimize performance with regard to the expression yield and the incorporation efficiency of amino acid precursors. A versatile screening platform based on batch-mode cell-free expression and central composite design was used. The performance of different extracts (S12 and S30), the concentration dependence of key components and the effect of different additives were investigated. We find that the initial expression yield can be enhanced twofold to threefold in this manner. The improved conditions comprise a modified S12 extract, optimized concentrations of creatine phosphate and key amino acids, as well as introduction of ketoacid additives. Our results show that current cell-free expression technology is far from optimal and that higher yields and increased utilization of the provided precursors are attainable with further optimization.
Authors:
Anders Pedersen; Kristofer Hellberg; Johan Enberg; B Göran Karlsson
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Publication Detail:
Type:  Journal Article     Date:  2010-07-13
Journal Detail:
Title:  New biotechnology     Volume:  28     ISSN:  1876-4347     ISO Abbreviation:  N Biotechnol     Publication Date:  2011 Apr 
Date Detail:
Created Date:  2011-04-12     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101465345     Medline TA:  N Biotechnol     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  218-24     Citation Subset:  IM    
Copyright Information:
Copyright © 2010 Elsevier B.V. All rights reserved.
Affiliation:
Swedish NMR Centre, University of Gothenburg, PO Box 465, SE-40530 Gothenburg, Sweden; Department of Chemistry, University of Gothenburg, PO Box 462, SE-40530 Gothenburg, Sweden.
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