Heterologous expression of the msp2 gene from Marasmius scorodonius.
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Issue Date
2009-05
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Show full item recordAbstract
For the heterologous expression of the msp2 gene from the edible mushroom Marasmius scorodonius in Escherichia coli the cDNA encoding the extracellular Msp2 peroxidase was cloned into the pBAD III expression plasmid. Expression of the protein with or without signal peptide was investigated in E. coli strains TOP10 and LMG194. Different PCR products were amplified for expression of the native target protein or a protein with a signal peptide. Omitting the native stop codon and adding six His-residues resulted in a fusion protein amenable to immune detection and purification by immobilised metal affinity chromatography. In E. coli the recombinant protein was produced in high yield as insoluble inclusion bodies. The influence of different parameters on MsP2 refolding was investigated. Active enzyme was obtained by glutathione-mediated oxidation in a medium containing urea, Ca(2+), and hemin.Citation
Heterologous expression of the msp2 gene from Marasmius scorodonius. 2009, 191 (5):397-402 Arch. Microbiol.Affiliation
Institut für Lebensmittelchemie, Leibniz Universität Hannover, Callinstrasse 5, Hannover, Germany.Journal
Archives of microbiologyPubMed ID
19247632Type
ArticleLanguage
enISSN
1432-072Xae974a485f413a2113503eed53cd6c53
10.1007/s00203-009-0462-2
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