Isolation and bioenergetic characterization of mitochondria from Pichia pastoris.
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.
González-Barroso
M Mar
MM
Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, Madrid, Spain. margb@cib.csic.es
Ledesma
Amalia
A
Lepper
Simone
S
Pérez-Magán
Elisa
E
Zaragoza
Pilar
P
Rial
Eduardo
E
D004734:Energy Metabolism; D005990:Glycerol / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism*; D009243:NAD / Q000378:metabolism; D010843:Pichia / Q000254:growth & development / Q000378:metabolism*
D009243:NAD; D005990:Glycerol