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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
1-2
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pubmed:dateCreated |
2004-4-5
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pubmed:abstractText |
The expression of recombinant trypsinogens from different mammalian origins in Escherichia coli typically leads to the formation of insoluble aggregates. This work describes the high level expression of human trypsinogen 1 in E. coli using the T7 expression system. Direct expression of trypsinogen was not possible, but the N-terminal fusion of the first 11 amino acids of the T7 protein 10 resulted in an expression level of 200 mg g(-1) bacterial dry mass. A refolding procedure was optimized, and a method using continuous feed of denatured product was developed. Thus the working concentration of trypsinogen could be raised four-fold, while the yield of active protein could be maintained at 20-35%. The refolded trypsinogen was converted to trypsin by autocatalytic activation, and the utility for the detachment of mammalian cells in culture was proven.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Apr
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pubmed:issn |
0168-1656
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
8
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pubmed:volume |
109
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
3-11
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pubmed:dateRevised |
2008-11-21
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pubmed:meshHeading |
pubmed-meshheading:15063609-Animals,
pubmed-meshheading:15063609-Base Sequence,
pubmed-meshheading:15063609-CHO Cells,
pubmed-meshheading:15063609-Capsid Proteins,
pubmed-meshheading:15063609-Cloning, Molecular,
pubmed-meshheading:15063609-Cricetinae,
pubmed-meshheading:15063609-Cricetulus,
pubmed-meshheading:15063609-Escherichia coli,
pubmed-meshheading:15063609-Genetic Vectors,
pubmed-meshheading:15063609-Inclusion Bodies,
pubmed-meshheading:15063609-Molecular Sequence Data,
pubmed-meshheading:15063609-Promoter Regions, Genetic,
pubmed-meshheading:15063609-Protein Denaturation,
pubmed-meshheading:15063609-Protein Folding,
pubmed-meshheading:15063609-Protein Renaturation,
pubmed-meshheading:15063609-Recombinant Fusion Proteins,
pubmed-meshheading:15063609-Trypsin,
pubmed-meshheading:15063609-Trypsinogen
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pubmed:year |
2004
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pubmed:articleTitle |
Bacterial expression and refolding of human trypsinogen.
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pubmed:affiliation |
Institute of Applied Microbiology, University of Natural Resources and Applied Life Sciences, Vienna, Austria.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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