Elsevier

Methods in Enzymology

Volume 55, 1979, Pages 144-148
Methods in Enzymology

[18] Preparation of Neurospora crassa mitochondria

https://doi.org/10.1016/0076-6879(79)55020-4Get rights and content

Publisher Summary

The fungus Neurospora crassa represents a eukaryotic cell with high biosynthetic activities. Cell mass doubles in 2–4 hours during exponential growth even in simple salt media with sucrose as the sole carbon source. The microorganism forms a mycelium of long hyphae during vegetative growth. The mitochondria can be isolated under relatively gentle conditions since a few breaks in the threadlike hyphae are sufficient to cause the outflow of the organelles. The chapter describes two methods for the physical disruption of the hyphae: (1) the cells are opened in a grind mill between two rotating corundum disks. This is a continuous and fast procedure and allows large- and small-scale preparations of mitochondria. (2) Hyphae are ground with sand in a mortar and pestle. This procedure can be applied to microscale preparations of mitochondria starting with minute amounts of cells. It also provides an overview on other procedures for the isolation of Neurospora mitochondria after the physical disruption or the enzymatic degradation of the cell wall.

References (19)

  • W. Sebald et al.

    FEBS Lett.

    (1968)
  • J. W. Greenawalt, D. O. Hall, and O. C. Wallis, this series,Vol. 10...
  • H.J. Vogel

    Microb. Genet. Bull.

    (1956)
  • R. H. Davis and F. J. de Serres, this series, Vol. 17A...
  • H. Weiss et al.

    Eur. J. Biochem.

    (1970)
  • G. von Jagow et al.

    Eur. J. Biochem.

    (1973)
  • A. von Ruecker et al.

    FEBS Lett.

    (1974)
  • H. Weiss et al.

    Eur. J. Biochem.

    (1971)
  • H. Weiss and W. Sebald, this series, Vol. 53...
There are more references available in the full text version of this article.

Cited by (32)

  • Biosynthesis of glycyrrhetinic acid 3-O-mono-β-d-glucuronide by free and immobilized Aspergillus terreus β-d-glucuronidase

    2011, Journal of Molecular Catalysis B: Enzymatic
    Citation Excerpt :

    Thereafter, the obtained cell homogenate was centrifuged at 5500 rpm for 15 min and the resulting supernatant “cell free extract” was lyophilized. The lyophilized material was used as the starting crude endocellular enzyme [19]. The lyophilized crude enzyme (120 g) was dissolved in 500 ml of 1 M Tris–maleate buffer (pH 6).

  • Cryo-electron tomography of Neurospora mitochondria

    2000, Journal of Structural Biology
View all citing articles on Scopus
View full text