Efficient expression systems for cysteine proteases of malaria parasites: Too good to be true?
Volume 4, Issue 2
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Pages 107 - 114http://dx.doi.org/10.4161/bioe.22348
: Escherichia coli
, Pichia pastoris
, auto-induction, codon optimization, cysteine proteases, efficient expression systems, malaria parasites
Authors: Emir Salas Sarduy and María de los A. Chávez Planes View affiliations
Papain-like cysteine proteases of malaria parasites are considered important chemotherapeutic targets or valuable models for the evaluation of drug candidates. Consequently, many of these enzymes have been cloned and expressed in Escherichia coli for their biochemical characterization. However, their expression has been problematic, showing low yield and leading to the formation of insoluble aggregates. Given that highly-productive expression systems are required for the high-throughput evaluation of inhibitors, we analyzed the existing expression systems to identify the causes of such apparent issues. We found that significant divergences in codon and nucleotide composition from host genes are the most probable cause of expression failure, and propose several strategies to overcome these limitations. Finally we predict that yeast hosts Saccharomyces cerevisiae and Pichia pastoris may be better suited than E. coli for the efficient expression of plasmodial genes, presumably leading to soluble and active products reproducing structural and functional characteristics of the natural enzymes.
ES Sarduy, AC Muñoz, SA Trejo, M de los A Chavéz Planes. High-level expression of Falcipain-2 in Escherichia coli by codon optimization and auto-induction. Protein Expr Purif 2012; 83: 59-69
PMID: 22450163 DOI: 10.1016/j.pep.2012.03.008
Received: August 1, 2012; Accepted: September 24, 2012; Published Online: September 27, 2012
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