To clarify the system for intracellular vitamin B_<12>(Cbl) transport in Euglena gracilis z, a unicellular eukaryote, Euglena Cbl-binding proteins were purified and characterized. Euglena cells contained numerous Cbl-binding proteins in addition to Cbl-dependent enzymes. These Cbl-binding proteins were shown to function as carriers of intracellular Cbl. Corrinoid specificities of cell growth, the Cbl uptake system and the binding proteins of E. gracilis were also determined. Aquacobalamin reductases catalyzing the reduction of aquacobalamin to cob(II)alamin in the synthesis of Cbl coenzymes were purified and characterized from Euglena mitochondria and rat liver. Although Euglena enzyme was specific to NADPH, NADH- and NADPH-linked enzymes occurred in both mitochondrial and microsomal membranes of rat liver. The Euglena enzyme was different from rat liver enzymes in the subcellular localization and immunological and enzymological properties. The Cbl-metabolism in Chlamydomonas reinhardtii, a photosynthesizing green alga, was studied to clarify physiological roles of Cbl in plant cells. The systems for intracellular Cbl transport and synthesis of Cbl coenzymes were also discussed in view of comparative biochemistry.
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