Subcellular Localization of the GABA‐Shunt Enzymes in Euglena gracilis Strain Z

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Abstract
Glutamate decarboxylase [EC 4.1.1.15], GABA-.alpha.-ketoglutarate aminotransferase and NAD-linked and NADP-linked succinic semialdehyde dehydrogenase [EC 1.2.1.16], all constituting the GABA-shunt pathway of glutamate metabolism are localized in the mitochondrial matrix in a streptomycin-bleached mutant of E. gracilis strain Z. Glutamate dehydrogenase [EC 1.4.1.2], requiring NADP as the cofactor, was distributed in the cytoplasm. An improved version of the controlled digestion method for preparing Euglena mitochondria, which involves use of trypsin and a trypsin inhibitor and removal of broken cells before mechanical disruption of cells, is also described.

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