Isolation, partial purification, and characterization of the cytochrome P-450-dependent monooxygenase system from the midgut of the earthworm Lumbricus terrestris.

Abstract

1. Cytochrome P-450 was purified from microsomes of the midgut of the earthworm Lumbricus terrestris up to a maximal specific content of 5.5 nmol P-450/mg protein. 2. At least 3 different cytochromes P-450 with apparent molecular weights of 48,000, 51,000 and 53,000 were identified by SDS-PAGE. 3. Western blot analysis with various polyclonal antibodies did not show structural epitopes common to the cytochromes P-450 of rodents or yeast and L. terrestris. 4. The microsomes contained about 43 pmol P-450/mg protein corresponding to 0.51 nmol P-450/g midgut and 64 pmol P-450/g body weight, respectively, and converted benzyloxyresorufin into resorufin with a Vmax of 2.12 pmol resorufin/min.mg protein and a Km of 770 nM benzyloxyresorufin at 25 degrees C, pH 8.0. 5. The microsomes exhibited a NADPH-cytochrome P-450 reductase activity of 9.4 nmol cytochrome c/min.mg protein. 6. The apparent molecular weight of the threefold-purified reductase was 63,000.

• Authors: A G Berghout, E Wenzel, J Büld, K J Netter
• Journal: Comparative biochemistry and physiology. C, Comparative pharmacology and toxicology (Comp Biochem Physiol C Comp Pharmacol Toxicol) Vol. 100 Issue 3 Pg. 389-96 ( 1991) ISSN: 0742-8413 [Print] England
• PMID: 1687532 (Publication Type: Journal Article)

Chemical References

• Cytochrome P-450 Enzyme System
• Oxygenases
• NADPH-Ferrihemoprotein Reductase

Topics

• Animals
• Cytochrome P-450 Enzyme System (chemistry, isolation & purification)
• Digestive System (enzymology)
• Microsomes (enzymology)
• NADPH-Ferrihemoprotein Reductase (isolation & purification)
• Oligochaeta (enzymology)
• Oxygenases (chemistry, isolation & purification)