Burkavidin: a novel secreted biotin-binding protein from the human pathogen Burkholderia pseudomallei.
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.
Institute of Inorganic Chemistry, Department of Chemistry, University of Basel, Spitalstrasse 51, 4056 Basel, Switzerland.
Alessia
A
Sardo
Therese
T
Wohlschlager
Cheikh
C
Lo
Heinz
H
Zoller
Thomas R
TR
Ward
Marc
M
Creus
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001710:Biotin / Q000378:metabolism*; D016957:Burkholderia pseudomallei / Q000737:chemistry; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002384:Catalysis; D003001:Cloning, Molecular; D004795:Enzyme Stability; D004926:Escherichia coli; D006801:Humans; D006864:Hydrogenase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007525:Isoelectric Focusing; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D055503:Protein Multimerization; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012238:Rhodium / Q000378:metabolism; D013237:Stereoisomerism; D019809:Streptavidin / Q000378:metabolism*
D001426:Bacterial Proteins; D002352:Carrier Proteins; D011994:Recombinant Proteins; C017071:biotin-binding proteins; D001710:Biotin; D019809:Streptavidin; D012238:Rhodium; D006864:Hydrogenase