Abstract
Background: A reliable, sensitive and practicable method for the measurement of intestinal lactase activity is needed. Method: The assay is based on the continuous measurement of released glucose by a coupled hexokinase/glucose-6-phosphate dehydrogenase (HK/G6PDH) reagent. Results: The procedure was first optimized for lactase from rat intestinal mucosa. The optimum pH is 6.5 and apparent Km was 17 mmol/l for lactose. The procedure was also adapted on a Cobas Mira automated analyzer; progress curves of the reaction rate can be continuously monitored. The automated assay correlated strongly with the conventional method of Dahlqvist (r<sup>2</sup> = 0.996). The described method has also been applied to human intestinal mucosa biopsies after determination of the catalytic properties of human enzyme (optimum pH 6.0 and apparent Km 34 mmol/l). Conclusion: The HK/G6PDH method is reliable, rapid, sensitive and easy to perform both manually as well as in the automated version. It is optimized for human and rat intestinal lactase.
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