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General methodology for isolation and characterization of Mitochondria from microorganisms

Lloyd, David 1979. General methodology for isolation and characterization of Mitochondria from microorganisms. Methods in Enzymology 55 (C) , pp. 135-144. 10.1016/0076-6879(79)55019-8

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This chapter discusses a survey of well-tested methods for the release of relatively undamaged mitochondria from eukaryotic microorganisms. The fragile protozoans may be disrupted in the absence of high liquid shearing forces by gentle hand homogenization or under more controlled conditions in a Chaikoff press. Organisms with rigid cell walls may be (a) ruptured mechanically by grinding or shaking with glass beads or (b) converted before breakage into spheroplasts by the use of cell-wall-degrading enzymes. Method (a) is rapid and is applicable to a wide variety of organisms. It can be employed for small-scale, multisample mitochondrial preparations or on a large scale, but it inevitably leads to some fragmentation of mitochondria and is better suited to the preparation of submitochondrial particles. Method (b) is more time-consuming and expensive but generally yields mitochondria of the highest quality. Its major disadvantage lies in its more limited applicability. Susceptibility to spheroplast formation by the action of the snail digestive enzyme varies markedly from one strain to another and depends on the stage or conditions of cell growth in the culture.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Biosciences
Publisher: Elsevier
ISSN: 0076-6879
Last Modified: 26 Mar 2020 16:46

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