Electrophoretically mediated microanalysis of a nicotinamide adenine dinucleotide-dependent enzyme and its facile multiplexing using an active pixel sensor UV detector.
An electrophoretically mediated microanalysis (EMMA) method has been developed for yeast alcohol dehydrogenase and quantification of reactant and product cofactors, NAD and NADH. The enzyme substrate ethanol (1% (v/v)) was added to the buffer (50 mM borate, pH 8.8). Results are presented for parallel capillary electrophoresis with a novel miniature UV area detector, with an active pixel sensor imaging an array of two or six parallel capillaries connected via a manifold to a single output capillary in a commercial CE instrument, allowing conversions with five different yeast alcohol dehydrogenase concentrations to be quantified in a single experiment.
Urban
Pawel L
PL
Department of Chemistry, University of York, Heslington, York YO10 5DD, UK.
Goodall
David M
DM
Bergström
Edmund T
ET
Bruce
Neil C
NC
D000426:Alcohol Dehydrogenase / Q000032:analysis* / Q000378:metabolism; D053801:Capillary Electrochromatography / Q000295:instrumentation / Q000379:methods*; D007527:Isoenzymes / Q000032:analysis / Q000378:metabolism; D008832:Microchemistry / Q000295:instrumentation / Q000379:methods*; D009243:NAD / Q000378:metabolism*; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D013997:Time Factors; D015003:Yeasts / Q000201:enzymology
D007527:Isoenzymes; D009243:NAD; D000426:Alcohol Dehydrogenase