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E. G. Romanova, A. A. Alekseeva, E. V. Pometun, V. I. Tishkov
Determination
of active site concentration and catalytic rate constant for recombinant
formate dehydrogenase from soya Glycine max
Abstract
Anaylsis of three-dimension structures of apo- and holo- formate plant
dehydrogenase (FDH) shows that binding of the enzyme with azide-ion and
coenzyme NAD+ in ternary complex should result in high quenching of
the enzyme fluorescence. Exitation and emission spectra indicated that
fluorescence is mainly due to tryptophan residues. Dependences of FDH
fluorescence on NAD+ and azide concentrations were studied. Using
these dependences the method for determination of enzyme active site
concentrations were developed and catalytic rate constant for recombinant FDH
from soya Glycine max was calculated.
Copyright (C) Chemistry Dept., Moscow State University, 2002
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