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Convenient procedure for the isolation of highly enriched, cytochrome P-450-containing microsomal fraction from Candida tropicalis

✍ Scribed by Othmar Käppeli; Marija Sauer; Armin Fiechter


Book ID
102627440
Publisher
Elsevier Science
Year
1982
Tongue
English
Weight
304 KB
Volume
126
Category
Article
ISSN
0003-2697

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✦ Synopsis


The suitability of Ca*' ions for the precipitation of the microsomal fraction from the hydrocarbon-grown yeast Candida tropicalis was evaluated. In the final procedure the microsomes were precipitated by the addition of 16 mM CIaClr. Crude extracts obtained from cells via spheroplast lysis were centrifuged at 12,OOOg for 15 min and at 25,OOOg for 15 min prior to precipitation. The cytochrome P-450 content of the fraction was between 0.22 and 0.35 nmol mg-' protein. The isolated microsomes exhibited both hexadecane hydroxylation activity and NADPH-cytochrome c reductase activity.