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The C subunit of Ideonella dechloratans chlorate reductase: expression, purification, refolding and heme reconstitution
Karlstad University, Faculty of Technology and Science, Department of Chemistry and Biomedical Sciences.
2005 (English)In: Protein Expression and Purification 41, 306-312Article in journal (Refereed)
Abstract [en]

The C subunit of Ideonella dechloratans chlorate reductase has been expressed in Escherichia coli as a GST fusion protein. Purification from inclusion bodies, followed by refolding and reconstitution with heme, produced a protein with a heme/protein ratio of 0.4, and with UVvis spectral characteristics similar to those of native chlorate reductase. Wavelength maxima for the Éø and É¿ bands in the reduced state were 559 and 529 nm for both native chlorate reductase and the reconstituted recombinant subunit, whereas the reduced Soret bands were found at 426 and 424 nm, respectively. These results support the notion of the C subunit as the cytochrome b moiety of I. dechloratans chlorate reductase. Moreover, the availability of a recombinant version of the C subunit is expected to facilitate further studies of electron transfer and protein interaction included in the reaction catalyzed by chlorate reductase

Place, publisher, year, edition, pages
Keyword [en]
Ideonella dechloratans, Chlorate, Chlorate reductase, ClrC, Heme, Expression, Reconstitution
National Category
Chemical Sciences
Research subject
URN: urn:nbn:se:kau:diva-24308OAI: diva2:598075
Available from: 2013-01-22 Created: 2013-01-22 Last updated: 2013-01-22

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Nilsson, Thomas
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