Uniformly oriented bacterial F0F1-ATPase immobilized on a semi-permeable membrane: a step towards biotechnological energy transduction.
The immobilization of F(0)F(1)-ATPase in uniform orientation is reported. The biotinylated and histidine-tagged subunits of the bacterial F(0)F(1)-ATPase complex were used for immobilization of the complex on artificial semi-permeable membranes resulting in 88+/-7.8 and 72+/-5.2% coupling of the enzymes. The immobilized enzymes retained over 90% activity. The immobilized ATPase/synthase was used for generation of ATP from ADP and P(i) at the expense of electrochemical potential energy. The re-usability, ratio of amount of enzyme immobilized to enzymic activity conferred on the membranes, ATP synthesized by assembled system and suitability of ATP generated for use in coupled enzymic reactions were determined.
Bhattacharya
Sumana
S
ABRD Company LLC, 1555 Wood Road, Cleveland, OH 44121, USA.
Schiavone
Marc
M
Nayak
Amiya
A
Bhattacharya
Sanjoy K
SK
D000244:Adenosine Diphosphate / Q000032:analysis / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D001709:Biotechnology; D019921:Biotinylation; D003545:Cysteine / Q000378:metabolism; D004735:Energy Transfer; D004926:Escherichia coli / Q000201:enzymology; D006639:Histidine / Q000737:chemistry; D008163:Luminescent Measurements; D008567:Membranes, Artificial; D010758:Phosphorus / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
D001426:Bacterial Proteins; D008567:Membranes, Artificial; D010761:Phosphorus Radioisotopes; D021122:Protein Subunits; D011994:Recombinant Proteins; D010758:Phosphorus; D006639:Histidine; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D006180:Proton-Translocating ATPases; D003545:Cysteine