Burkavidin: A Novel Secreted Biotin-Binding Protein from the Human Pathogen Burkholderia Pseudomallei
Protein Expression Purif. 2011, 77, 131-139, 10.1016/j.pep.2011.01.003
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.
Metal: RhLigand type: DiphenylphosphineHost protein: BurkavidinAnchoring strategy: SupramolecularOptimization: Chemical & geneticReaction: HydrogenationMax TON: ~110ee: 65PDB: ---Notes: ---
Design and Evolution of Artificial Metalloenzymes: Biomimetic AspectsReview
Prog. Inorg. Chem. 2011, 203-253, 10.1002/9781118148235.ch4