1. Burkavidin: a novel secreted biotin-binding protein from the human pathogen Burkholderia pseudomallei.
- Author
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Sardo A, Wohlschlager T, Lo C, Zoller H, Ward TR, and Creus M
- Subjects
- Amino Acid Sequence, Bacterial Proteins chemistry, Bacterial Proteins genetics, Burkholderia pseudomallei chemistry, Carrier Proteins chemistry, Carrier Proteins genetics, Catalysis, Cloning, Molecular, Enzyme Stability, Escherichia coli, Humans, Hydrogenase chemistry, Hydrogenase genetics, Isoelectric Focusing, Kinetics, Models, Molecular, Molecular Sequence Data, Protein Multimerization, Recombinant Proteins chemistry, Recombinant Proteins genetics, Rhodium metabolism, Stereoisomerism, Bacterial Proteins metabolism, Biotin metabolism, Carrier Proteins metabolism, Hydrogenase metabolism, Recombinant Proteins metabolism, Streptavidin metabolism
- Abstract
The avidin-biotin technology has many applications, including molecular detection; immobilization; protein purification; construction of supramolecular assemblies and artificial metalloenzymes. Here we present the recombinant expression of novel biotin-binding proteins from bacteria and the purification and characterization of a secreted burkavidin from the human pathogen Burkholderia pseudomallei. Expression of the native burkavidin in Escherichia coli led to periplasmic secretion and formation of a biotin-binding, thermostable, tetrameric protein containing an intra-monomeric disulphide bond. Burkavidin showed one main species as measured by isoelectric focusing, with lower isoelectric point (pI) than streptavidin. To exemplify the potential use of burkavidin in biotechnology, an artificial metalloenzyme was generated using this novel protein-scaffold and shown to exhibit enantioselectivity in a rhodium-catalysed hydrogenation reaction., (Copyright © 2011 Elsevier Inc. All rights reserved.)
- Published
- 2011
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