Abstract
This chapter focuses on the cytochrome oxidase obtained from thermophilic bacteria PS3. The thermophilic bacterium PS3 is a Gram-positive spore former. It produces a-, b-, and c-type cytochromes. Under highly aerobic conditions, the terminal oxidase is an aa3-type cytochrome oxidase, but under air-restricted conditions cytochrome o replaces cytochrome aa3. From cells grown under highly aerated conditions, aa3-type cytochrome oxidase can be prepared by following the α-band spectrum of the reduced form. The chapter describes a freeze-thaw method for reconstituting beef heart cytochrome oxidase and Thermus thermophilus cytochrome oxidase into vesicles. The cholate-deoxycholate dialysis method was not good for reconstitution of PS3 cytochrome oxidase. Two or three cycles of the freeze-thaw sonication gave the highest H+ pumping activity and a slow leakage. PS3 cytochrome oxidase vesicles thus prepared pump H+ upon addition of reduced cytochrome c under aerobic conditions.
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