A microplate assay for d-xylose/ d-glucose isomerase

A modification of the resorcinol method of Kulka 1 for the determination of ketoses is described. Though being a stopped enzyme test, it is much more sensitive than the carbazol method and by applying microtiter plates and measuring with an ELISA reader, a large number of tests can be performed with...

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Bibliographic Details
Published in:Enzyme and microbial technology 1998, Vol.22 (8), p.721-723
Main Authors: Schenk, Martin, Bisswanger, Hans
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
d-Xylose isomerase
Enzyme engineering
enzyme test
fructose determination
Fundamental and applied biological sciences. Psychology
Methods. Procedures. Technologies
microplate
Miscellaneous
resorcinol
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Summary:A modification of the resorcinol method of Kulka 1 for the determination of ketoses is described. Though being a stopped enzyme test, it is much more sensitive than the carbazol method and by applying microtiter plates and measuring with an ELISA reader, a large number of tests can be performed within a short time, thereby facilitating initial velocity studies. The test is linear up to a concentration of 2.5 m m d-xylulose even in the presence of 10 m m d-xylose and 2 m m d-fructose in the presence of 10 m m d-glucose. The sensitivity is 25 μ m for xylulose and 38 μ m for fructose. The test method is insensitive to perturbations of substances frequently used in isolation procedures such as ammonium sulfate, Triton X-100, PEG 6000, sodium dodecyl sulfate, and ethanol in moderate concentrations.
ISSN:0141-0229
1879-0909
DOI:10.1016/S0141-0229(98)00003-9