An improved method of photometric determination of cyclodextrin glucanotransferase activity |
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Authors: | N. G. Usanov E. A. Gil’vanova P. A. Elizar’ev E. A. Prutsakova A. I. Melent’ev |
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Affiliation: | (1) Institute of Biology, Ufa Scientific Center, Russian Academy of Sciences, Ufa, 450054, Bashkortostan, Russia |
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Abstract: | The method of spectrophotometric determination of cyclodextrin glucanotransferase (CGTase, EC 2.4.1.19) activity with the use of phenolphthalein as a colored reagent has been improved. This technique includes an enzymatic reaction at 40°C for 60 min in 2% starch, with subsequent supplementation of the reaction mixture (0.5 ml) with the phenolphthalein reagent (3.0 ml) prepared in 0.1 M potassium carbonate buffer (pH 11.0) according to a special procedure, and measurement of the optical density of the obtained mixture at 553 nm. The activity was calculated using the exponential growth equation that connects a drop in the optical density and the degree of dilution of the enzyme. The described technique is suitable for working in a sufficiently broad range of specific activity of β-CGTase and does not require precise adjustment of the degree of dilution of solutions analyzed. |
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