Fluorometric determination of isocitrate dehydrogenase (EC 1.1.1.42; 1; NADP+ dependent) in ruminant milk

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Larsen, T. | Rupp, R. | Friggens, Nicolas, N.C. | Pires, J.A.A.

Edité par CCSD ; Published by Elsevier (since 2021) / Cambridge University Press (until 2020) -

International audience. The enzyme isocitrate dehydrogenase (EC 1.1.1.42; 1; NADP+ dependent) located in the mammary cell cytosol mediates the synthesis of the majority of reducing equivalents for the energetically demanding milk fat and cholesterol synthesis in mammary cell cytosol. The present article presents a novel fluoro-metric method for quantification of the activity of this enzyme (IDH) in ruminant milk without pretreat-ment of the sample. Further, 493 goat milk samples - harvested before, during and after a nutritional restriction - were analysed for IDH activity i) with addition of extra substrate (isocitrate), and ii) with the intrinsic isocitrate solely. The IDH activity ranged from 0.22 to 15.4 units [nano moles product/ (ml * min)] (un-supplemented) and from 0.22 to 45.6 units (isocitrate supplemented). The IDH activity increased considerably in milk during the nutritional restriction period concomitant with the increase in the metabolite isocitrate concentration and somatic cell count and returned to the initial level shortly after restriction period. The present 'high through-put' analytical method may be beneficial in future studies to phenotype modifications in mammary energy metabolism and milk fat synthesis, for which IDH activity may be a biomarker. (c) 2022 The Author(s). Published by Elsevier B.V. on behalf of The Animal Consortium. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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