A spectrophotometric assay for 6-phosphogluconolactonase involving the use of immobilized enzymes to prepare the labile 6-phosphoglucono-delta-lactone substrate. |
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| Authors: | R D Moir and G B Stokes |
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| Affiliation: | Department of Biochemistry, University of Western Australia, Perth, Australia. |
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| Abstract: | We report the development of a new spectrophotometric assay for 6-phosphogluconolactonase. The labile substrate 6-phosphoglucono-delta-lactone is prepared from glucose 6-phosphate by enzymes co-immobilized on Sepharose beads. The assay has the advantages of high sensitivity for routine determination of enzyme activity and allows determination of both Km and Vmax. from a single assay. A method for estimating the contribution of spontaneous hydrolysis to total hydrolysis is described. This assay overcomes the problems encountered with all previous assays. |
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