TY - JOUR
T1 - Disulfide bond formation and activation of Escherichia coli β-galactosidase under oxidizing conditions
AU - Seras-Franzoso, Joaquin
AU - Affentranger, Roman
AU - Ferrer-Navarro, Mario
AU - Daura, Xavier
AU - Villaverde, Antonio
AU - García-Fruitósa, Elena
PY - 2012/4/1
Y1 - 2012/4/1
N2 - Escherichia coli β-galactosidase is probably the most widely used reporter enzyme in molecular biology, cell biology, and biotechnology because of the easy detection of its activity. Its large size and tetrameric structure make this bacterial protein an interesting model for crystallographic studies and atomic mapping. In the present study, we investigate a version of Escherichia coli β-galactosidase produced under oxidizing conditions, in the cytoplasm of an Origami strain. Our data prove the activation of this microbial enzyme under oxidizing conditions and clearly show the occurrence of a disulfide bond in the β-galactosidase structure. Additionally, the formation of this disulfide bond is supported by the analysis of a homology model of the protein that indicates that two cysteines located in the vicinity of the catalytic center are sufficiently close for disulfide bond formation. © 2012, American Society for Microbiology.
AB - Escherichia coli β-galactosidase is probably the most widely used reporter enzyme in molecular biology, cell biology, and biotechnology because of the easy detection of its activity. Its large size and tetrameric structure make this bacterial protein an interesting model for crystallographic studies and atomic mapping. In the present study, we investigate a version of Escherichia coli β-galactosidase produced under oxidizing conditions, in the cytoplasm of an Origami strain. Our data prove the activation of this microbial enzyme under oxidizing conditions and clearly show the occurrence of a disulfide bond in the β-galactosidase structure. Additionally, the formation of this disulfide bond is supported by the analysis of a homology model of the protein that indicates that two cysteines located in the vicinity of the catalytic center are sufficiently close for disulfide bond formation. © 2012, American Society for Microbiology.
U2 - https://doi.org/10.1128/AEM.06923-11
DO - https://doi.org/10.1128/AEM.06923-11
M3 - Article
VL - 78
SP - 2376
EP - 2385
ER -