SUMMARY: A cyclic pathway of NADPH generation involving interconversion of mannitol and fructose has been proposed to occur in fungi. In three enzymes of this proposed mannitol cycle (hexokinase, NADP-mannitol dehydrogenase and mannitol-1-phosphate phosphatase) were shown to be localized exclusively in the cytosol. Two isoenzymes of the fourth enzyme (mannitol-1-phosphate dehydrogenase) were detected and shown to be localized respectively in the mitochondrion and the cytosol. The mitochondrial isoenzyme appeared to be present on the outer face of the inner mitochondrial membrane. No evidence was found for a coordinated change in the maximal activities of the enzymes of the proposed mannitol cycle in extracts prepared from mycelia grown on six different carbon, and three different nitrogen sources nor for any increase in these activities induced by growth on NO-. Studies of this type in which other NADP-linked dehydrogenases were measured showed that for most carbon sources tested growth on NO- increased the maximal activity of NADP-isocitrate dehydrogenase as well as that of glucose-6-phosphate and 6-phosphogluconate dehydrogenases but had little effect on the maximal activity of NADP-malate dehydrogenase (decarboxylating). Our studies provide no support for the operation of the mannitol cycle, or for the proposed role of this cycle in NADPH generation in


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