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J Biol Chem, Vol. 274, Issue 10, 6212-6218, March 5, 1999

Functional Expression of the Plant Alternative Oxidase Affects Growth of the Yeast Schizosaccharomyces pombe

From the Department of Biochemistry, School of Biological Sciences, University of Sussex, Falmer, Brighton BN1 9QG, United Kingdom and the ^§ IMBW Section Microbiology, Vrije Universiteit, De Boelelaan 1087, 1081 HV Amsterdam, The Netherlands

We have investigated the extent to which functional expression of the plant alternative oxidase (from Sauromatum guttatum) in Schizosaccharomyces pombe affects yeast growth. When cells are cultured on glycerol, the maximum specific growth rate is decreased from 0.13 to 0.11 h¹ while growth yield is lowered by 20% (from 1.14 × 10⁸ to 9.12 × 10⁷ cells ml¹). Kinetic studies suggest that the effect on growth is mitochondrial in origin. In isolated mitochondria we found that the alternative oxidase actively competes with the cytochrome pathway for reducing equivalents and contributes up to 24% to the overall respiratory activity. Metabolic control analysis reveals that the alternative oxidase exerts a considerable degree of control (22%) on total electron flux. Furthermore, the negative control exerted by the alternative oxidase on the flux ratio of electrons through the cytochrome and alternative pathways is comparable with the positive control exerted on this flux-ratio by the cytochrome pathway. To our knowledge, this is the first paper to report a phenotypic effect because of plant alternative oxidase expression. We suggest that the effect on growth is the result of high engagement of the non-protonmotive alternative oxidase in yeast respiration that, consequently, lowers the efficiency of energy conservation and hence growth.

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