Successful recombinant production of Allochromatium vinosum cytochrome c' requires coexpression of cmm genes in heme-rich Escherichia coli JCB712

T.H. Evers, M. Merkx

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9 Citaten (Scopus)

Samenvatting

Cytochrome c' from the purple photosynthetic bacterium Allochromatium vinosum (CCP) displays a unique, reversible dimer-to-monomer transition upon binding of NO, CO, and CN-. This small, four helix bundle protein represents an attractive model for the study of other heme protein biosensors, provided a recombinant expression system is available. Here we report the development of an efficient expression system for CCP that makes use of a maltose binding protein fusion strategy to enhance periplasmic expression and allow easy purifn. by affinity chromatog. Coexpression of cytochrome c maturase genes and the use of a heme-rich Escherichia coli strain were found to be necessary to obtain reasonable yields of cytochrome c'. Characterization using CD, UV-vis spectroscopy, and size-exclusion chromatog. confirms the native-like properties of the recombinant protein, including its ligand-induced monomerization.
Originele taal-2Engels
Pagina's (van-tot)668-674
TijdschriftBiochemical and Biophysical Research Communications
Volume327
Nummer van het tijdschrift3
DOI's
StatusGepubliceerd - 2005

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