Fluorescence and chemical reactivity as empirical indices of matrix-induced alterations in immobilized enzymes.
The fluorescence emission and chemical reactivity of soluble and Sepharose-bound leucine aminopeptidase and carboxypeptidase A were investigated and the results compared. Both natural fluorescence and the fluorescence of fluorescein-labeled enzymes were utilized. The front-surface viewing technique enables us to measure the apparent fluorescence quantum yields and the rotational relaxation times of fluorescent groups within native unbound and carrier-fixed enzymes. The chemical reactivity was probed with Ellman's reagent and iodoacetic acid. Both types of methods were found to be useful in detection and quantitation of matrix-induced alterations in immobilized enzymes.
D002268:Carboxypeptidases / Q000378:metabolism*; D005452:Fluoresceins; D005453:Fluorescence; D007461:Iodoacetates / Q000378:metabolism; D007930:Leucine; D007931:Leucyl Aminopeptidase / Q000378:metabolism*; D012995:Solubility; D014364:Tryptophan / Q000378:metabolism
D005452:Fluoresceins; D007461:Iodoacetates; D014364:Tryptophan; D002268:Carboxypeptidases; D007931:Leucyl Aminopeptidase; D007930:Leucine