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Carbonic anhydrase (CA), a well-characterized metalloenzyme, is associated with oxygen-18 (  18 O)-isotopic fractionations of CO 2 . To investigate how CA activity links the  18 O of breath CO 2  to pre-diabetes (PD) and type 2 diabetes (T2D) during metabolism, we studied pre- and post-dose CA activities in erythrocytes with simultaneous monitoring of  18 O/  16 O-isotope ratios of breath CO 2  and thereafter elucidated potential metabolic pathways underlying CA alteration in the pathogenesis of T2D. Here we show that the post-dose CA activity in both T2D and PD was markedly enhanced, whereas the non-diabetic controls (NDC) exhibited a considerable reduction in post-dose CA activity when compared with their basal CA activities. However, T2D and PD exhibited isotopic enrichments of  18 O in breath CO 2 , while a marked depletion of  18 O in CO 2  was manifested in NDC. Thus, the isotopic enrichments and depletions of  18 O in breath CO 2  were well correlated with the changes in CA activities for controls, PD and T2D. Our findings suggest the changes in CA activities in erythrocytes may contribute to the pathogenesis of T2D and the breath C  18 O  16 O regulated by the CA activity as a potential biomarker for non-invasive assessment of T2D, and thus may open a new method for treating T2D.

Oxygen-18 isotope of breath CO2 linking to erythrocytes carbonic anhydrase activity: a biomarker for pre-diabetes and type 2 diabetes