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Isolation and bioenergetic characterization of mitochondria from Pichia pastoris
Author(s) -
Mar GonzálezBarroso M.,
Ledesma Amalia,
Lepper Simone,
PérezMagán Elisa,
Zaragoza Pilar,
Rial Eduardo
Publication year - 2006
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.1355
Subject(s) - pichia pastoris , bioenergetics , yeast , biology , mitochondrion , alternative oxidase , pichia , biochemistry , saccharomyces cerevisiae , gene , recombinant dna
Pichia pastoris is a methylotrophic yeast of high biotechnological interest. The bioenergetic properties of mitochondria from Pichia pastoris have not yet been determined. We report on a protocol for the isolation of the mitochondria in a state that shows good energy coupling. Analysis of Pichia pastoris growth and bioenergetic properties of the isolated mitochondria reveals that glycerol is the carbon source that yields the best results. Under our growth conditions, mitochondria oxidize external NADH but do not possess an alternative oxidase. Finally, Pichia pastoris mitochondria also lack the nucleotide‐stimulated uncoupling pathway previously identified in Saccharomyces cerevisiae . Copyright © 2006 John Wiley & Sons, Ltd.