Notable increase in content of cytochrome P-450 enzymes in Saccharomyces cerevisiae in repetitive alternating culturing

1995 ◽  
Vol 23 (3) ◽  
pp. 499S-499S
Author(s):  
Michael Winkler ◽  
Bryan Underwood ◽  
Alan Wiseman
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Notable Increase ◽  
Cytochrome P 450

Isolation of a cytochrome P-450 structural gene from Saccharomyces cerevisiae

Gene ◽  
1986 ◽  
Vol 45 (3) ◽  
pp. 237-245 ◽  
Author(s):  
V.F. Kalb ◽  
J.C. Loper ◽  
C.R. Dey ◽  
C.W. Woods ◽  
T.R. Sutter
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Structural Gene ◽  
Cytochrome P 450

scholarly journals Characterization of Saccharomyces cerevisiae CYP51 and a CYP51 fusion protein with NADPH cytochrome P-450 oxidoreductase expressed in Escherichia coli.

1997 ◽  
Vol 41 (4) ◽  
pp. 776-780 ◽  
Author(s):  
K Venkateswarlu ◽  
D E Kelly ◽  
S L Kelly
Keyword(s):  
Escherichia Coli ◽  
Saccharomyces Cerevisiae ◽  
Reductase Activity ◽  
Spectral Response ◽  
Cumene Hydroperoxide ◽  
Type Ii ◽  
Nadph Cytochrome ◽  
Binding Spectra ◽  
Cytochrome P 450 ◽  
Fus Protein

Saccharomyces cerevisiae CYP51, target of azole antifungal agents, and CYP51 fused with S. cerevisiae cytochrome P-450 oxidoreductase (FUS protein) were expressed in active forms in Escherichia coli by cloning into pET15b. The expression was monitored immunologically, catalytically, and by using reduced carbon monoxide difference and type II binding spectra. CYP51 and FUS enzymes were located in membranes and produced a Soret peak at 448 nm in the reduced CO difference spectrum. The cytochrome P-450 contents in the membrane fractions containing CYP51 and FUS proteins were 12.8 +/- 2.6 and 17.4 +/- 3.7 pmol/mg of protein, respectively. The NADPH cytochrome P-450 oxidoreductase (CPR) content was estimated to be 15.7 +/- 1.1 pmol/mg of protein in FUS membrane fractions. FUS protein catalyzed the demethylation of substrate at the 14alpha position, with a turnover number of 1.96 +/- 0.37 min(-1) in the presence of NADPH. No reductase activity was observed in membrane fractions containing CYP51, and therefore, CYP51 did not function catalytically in the presence of NADPH, but in the presence of an artificial electron donor, cumene hydroperoxide, activity was comparable to that of the FUS enzyme. Further support for a normal structure for the hemoproteins was obtained from type II binding spectra, in which the spectral response was saturated with an equimolar concentration of ketoconazole.

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