Information on EC 1.14.14.154 - sterol 14alpha-demethylase and Organism(s) Saccharomyces cerevisiae and UniProt Accession P10614

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lanosterol 14-demethylase

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14-demethylase

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14alpha-demethylase

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14alpha-methylsterol 14alpha-demethylase

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14alpha-sterol demethylase

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14DM

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CYP51

CYPL1

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cytochrome CYP51

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cytochrome P 450 CYP51

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cytochrome P-450 lanosterol 14alpha-demethylase

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cytochrome P-450-dependent 14alpha-sterol demethylase

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cytochrome P-450-dependent obtusifoliol 14alpha-demethylase

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cytochrome P-450/14DM

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cytochrome P-45014DM

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cytochrome P450 14DM

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cytochrome P450 51

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cytochrome P450 CYP51

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cytochrome-P450 14alpha-demethylase

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demethylase, methylsterol 14alpha-

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eburicol 14 alpha-demethylase

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eburicol 14alpha-demethylase

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ERG11

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Erg11p

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672337, 701727

lanosterol 14 alpha-demethylase

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lanosterol 14-demethylase

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lanosterol 14alpha-demethylase

lanosterol 14alpha-methyldemethylase

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lanosterol C-14 demethylase

lanosterol demethylase

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LDM

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methylsterol 14alpha-demethylase (P 450 CYP51)

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Obtusifoliol 14-alpha demethylase

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obtusifoliol 14-demethylase

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obtusifoliol 14alpha-demethylase

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obtusifoliol-metabolizing 14alpha-demethylase

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obtusufoliol 14-demethylase

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P 450 lanosterol C-14 demethylase

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P-450 lanosterol demethylase

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P-45014DM

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P-45014DM-containing monooxygenase system

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P-450OBT 14DM

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P450(14DM)

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P450-14DM

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P450-L1A1

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P45014DM

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sterol 14-demethylase

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sterol 14-demethylase P450

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sterol 14alpha-demethylase

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sterol 14alpha-demethylase (CYP51)

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sterol 14alpha-demethylase cytochrome P 450

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sterol C14 demethylase

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additional information

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the enzyme belongs to the CYP51 family

a 14alpha-methylsteroid + 3 [reduced NADPH-hemoprotein reductase] + 3 O2 = a DELTA14-steroid + formate + 3 [oxidized NADPH-hemoprotein reductase] + 4 H2O

Biosynthesis of secondary metabolites, Steroid biosynthesis

oxygenation

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redox reaction

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oxidation

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reduction

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hydroxylation

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monooxygenation

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BRENDA

cholesterol biosynthesis

KEGG

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-, -, -, -, -, -, -, -

sterol,[reduced NADPH-hemoprotein reductase]:oxygen oxidoreductase (14-methyl cleaving)

This cytochrome P-450 (heme-thiolate) enzyme acts on a range of steroids with a 14alpha-methyl group, such as obtusifoliol and lanosterol. The enzyme catalyses a hydroxylation and a reduction of the 14alpha-methyl group, followed by a second hydroxylation, resulting in the elimination of formate and formation of a 14(15) double bond.

138674-19-8

deleted registry number

341989-59-1

deleted registry number

60063-87-8

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90463-45-9

deleted registry number

lanosterol + NADPH + O2

4,4-dimethylcholesta-8,14,24-trien-3-ol + NADP+ + H2O

artificial fused enzymes have comparable activity to the reconstituted system. Reduction of CYP51 both in the fused enzyme and the reconstituted system is biphasic and consisted of an initial fast phase followed by a slow phase

-

?

24,25-dihydrolanosterol + [reduced NADPH-hemoprotein reductase] + O2

4,4-dimethyl-5alpha-cholesta-8,14-dien-3beta-ol + formate + [oxidized NADPH-hemoprotein reductase] + H2O

24,25-dihydrolanosterol + [reduced NADPH-hemoprotein reductase] + O2

?

24,28-dihydroobtusifoliol + [reduced NADPH-hemoprotein reductase] + O2

4alpha-methyl-5alpha-ergosta-8,14-dien-3beta-ol + formate + [oxidized NADPH-hemoprotein reductase] + H2O

24-methylene-24,25-dihydrolanosterol + [reduced NADPH-hemoprotein reductase] + O2

4,4-dimethyl-ergosta-8,14,24(28)-trien-3beta-ol + formate + [oxidized NADPH-hemoprotein reductase] + H2O

24-methylenedihydrolanosterol + [reduced NADPH-hemoprotein reductase] + O2

?

-

-

?

7-lanosten-3beta-ol + [reduced NADPH-hemoprotein reductase] + O2

4,4-dimethylcholesta-7,14-dien-3beta-ol + formate + [oxidized NADPH-hemoprotein reductase] + H2O

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very low activity

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?

7-lanostene-3beta,32-diol + [reduced NADPH-hemoprotein reductase] + O2

4,4-dimethylcholesta-7,14-dien-3beta-ol + formate + [oxidized NADPH-hemoprotein reductase] + H2O

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very low activity

-

?

8-lanostene-3beta,32-diol + [reduced NADPH-hemoprotein reductase] + O2

4,4-dimethylcholesta-8,14-dien-3beta-ol + formate + [oxidized NADPH-hemoprotein reductase] + H2O

-

-

?

lanosterol + [reduced NADPH-hemoprotein reductase] + O2

4,4-dimethyl-5alpha-cholesta-8,14,24-trien-3beta-ol + formate + [oxidized NADPH-hemoprotein reductase] + H2O

6 entries

lanosterol + [reduced NADPH-hemoprotein reductase] + O2

4,4-dimethylcholesta-8,14,24-trienol + formate + [oxidized NADPH-hemoprotein reductase] + H2O

lanosterol + [reduced NADPH-hemoprotein reductase] + O2

?

-

-

?

lanosterol + [reduced NADPH-hemoprotein reductase] + O2

? + [oxidized NADPH-hemoprotein reductase] + H2O

-

-

?

obtusifoliol + [reduced NADPH-hemoprotein reductase] + O2

4alpha-methyl-5alpha-ergosta-8,14,24(28)-trien-3beta-ol + formate + [oxidized NADPH-hemoprotein reductase] + H2O

additional information

?

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24 entries

24,25-dihydrolanosterol + [reduced NADPH-hemoprotein reductase] + O2

4,4-dimethyl-5alpha-cholesta-8,14-dien-3beta-ol + formate + [oxidized NADPH-hemoprotein reductase] + H2O

24,25-dihydrolanosterol + [reduced NADPH-hemoprotein reductase] + O2

?

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the enzyme is involved in ergosterol and cholesterol biosynthesis

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?

lanosterol + [reduced NADPH-hemoprotein reductase] + O2

4,4-dimethyl-5alpha-cholesta-8,14,24-trien-3beta-ol + formate + [oxidized NADPH-hemoprotein reductase] + H2O

lanosterol + [reduced NADPH-hemoprotein reductase] + O2

4,4-dimethylcholesta-8,14,24-trienol + formate + [oxidized NADPH-hemoprotein reductase] + H2O

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deletion of the enzyme is lethal

-

?

lanosterol + [reduced NADPH-hemoprotein reductase] + O2

?

-

-

?

additional information

?

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9 entries

cytochrome P450

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285344, 285345, 285346, 285347, 285348, 285351, 285352, 285354

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heme

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heme-thiolate enzyme

285344, 285345, 285346, 285347, 285348, 285351, 285352, 285354

ketoconazole

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24,25-dihydrolanosterol

24-methylene-24,25-dihydrolanosterol

anti-P-45014DM antibodies

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complete inhibition

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azole antifungal agents

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specific inhibitors

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clotrimazole

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CO

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ratio CO:O2 of 90:10, 51.1% inhibition, ratio CO:O2 of 95:5, 100% inhibition

difenoconazole

whole-cell antifungal activity of both the R- and S-enantiomers of tebuconazole, prothioconazole, prothioconazole-desthio, and oxo-prothioconazole as well as for fluquinconazole, prochloraz and a racemic mixture of difenoconazole are determined. In vitro binding studies with the affinity purified enzyme are used to show tight type II binding to the yeast enzyme for all compounds tested except prothioconazole and oxo-prothioconazole. Comparison with CYP51 structures from fungal pathogens including Candida albicans, Candida glabrata and Aspergillus fumigatus provides strong evidence for a highly conserved CYP51 structure including the drug binding site

econazole

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fluconazole

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fluquinconazole

whole-cell antifungal activity of both the R- and S-enantiomers of tebuconazole, prothioconazole, prothioconazole-desthio, and oxo-prothioconazole as well as for fluquinconazole, prochloraz and a racemic mixture of difenoconazole are determined. In vitro binding studies with the affinity purified enzyme are used to show tight type II binding to the yeast enzyme for all compounds tested except prothioconazole and oxo-prothioconazole. Comparison with CYP51 structures from fungal pathogens including Candida albicans, Candida glabrata and Aspergillus fumigatus provides strong evidence for a highly conserved CYP51 structure including the drug binding site

imidazole inhibitors

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itraconazole

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ketoconazole

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lanosterol

menadione

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0.125 mM: 62.1% inhibition

Metyrapone

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strong inhibition, 0.1 mM: 57.3% inhibition

miconazole

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nitrogen

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nitrogen atmosphere

obtusifoliol

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24.4% inhibition of 24,25-dihydrolanosterol, DHL, demethylation, no inhibition of lanosterol and 24-methylene-24,25-dihydrolanosterol demethylation

oxo-prothioconazole

whole-cell antifungal activity of both the R- and S-enantiomers of tebuconazole, prothioconazole, prothioconazole-desthio, and oxo-prothioconazole as well as for fluquinconazole, prochloraz and a racemic mixture of difenoconazole are determined. In vitro binding studies with the affinity purified enzyme are used to show tight type II binding to the yeast enzyme for all compounds tested except prothioconazole and oxo-prothioconazole. Comparison with CYP51 structures from fungal pathogens including Candida albicans, Candida glabrata and Aspergillus fumigatus provides strong evidence for a highly conserved CYP51 structure including the drug binding site

Prochloraz

whole-cell antifungal activity of both the R- and S-enantiomers of tebuconazole, prothioconazole, prothioconazole-desthio, and oxo-prothioconazole as well as for fluquinconazole, prochloraz and a racemic mixture of difenoconazole are determined. In vitro binding studies with the affinity purified enzyme are used to show tight type II binding to the yeast enzyme for all compounds tested except prothioconazole and oxo-prothioconazole. Comparison with CYP51 structures from fungal pathogens including Candida albicans, Candida glabrata and Aspergillus fumigatus provides strong evidence for a highly conserved CYP51 structure including the drug binding site

prothioconazole

whole-cell antifungal activity of both the R- and S-enantiomers of tebuconazole, prothioconazole, prothioconazole-desthio, and oxo-prothioconazole as well as for fluquinconazole, prochloraz and a racemic mixture of difenoconazole are determined. In vitro binding studies with the affinity purified enzyme are used to show tight type II binding to the yeast enzyme for all compounds tested except prothioconazole and oxo-prothioconazole. Comparison with CYP51 structures from fungal pathogens including Candida albicans, Candida glabrata and Aspergillus fumigatus provides strong evidence for a highly conserved CYP51 structure including the drug binding site

prothioconazole-desthio

whole-cell antifungal activity of both the R- and S-enantiomers of tebuconazole, prothioconazole, prothioconazole-desthio, and oxo-prothioconazole as well as for fluquinconazole, prochloraz and a racemic mixture of difenoconazole are determined. In vitro binding studies with the affinity purified enzyme are used to show tight type II binding to the yeast enzyme for all compounds tested except prothioconazole and oxo-prothioconazole. Comparison with CYP51 structures from fungal pathogens including Candida albicans, Candida glabrata and Aspergillus fumigatus provides strong evidence for a highly conserved CYP51 structure including the drug binding site

SKF-525A

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potent inhibitor, 1.0 mM: 100% inhibition

tebuconazole

whole-cell antifungal activity of both the R- and S-enantiomers of tebuconazole, prothioconazole, prothioconazole-desthio, and oxo-prothioconazole as well as for fluquinconazole, prochloraz and a racemic mixture of difenoconazole are determined. In vitro binding studies with the affinity purified enzyme are used to show tight type II binding to the yeast enzyme for all compounds tested except prothioconazole and oxo-prothioconazole. Comparison with CYP51 structures from fungal pathogens including Candida albicans, Candida glabrata and Aspergillus fumigatus provides strong evidence for a highly conserved CYP51 structure including the drug binding site

triazole inhibitors

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voriconazole

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displays the greatest specificity

additional information

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7-lanostene-3beta,32-diol

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activation

7-lanostene-3beta-ol

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activation

8-lanostene-3beta,32-diol

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activation

8-lanostene-3beta-ol

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activation

Cycloartenol

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activation of enzymatic reaction

lanosterol

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induction and activation

0.004 - 0.005

lanosterol

0.017 - 0.02

24,25-dihydrolanosterol

0.0077 - 0.0087

24-methylene-24,25-dihydrolanosterol

0.003

7-lanostene-3beta,32-diol

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0.0001

8-lanostene-3beta,32-diol

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0.005 - 0.0067

lanosterol

0.012

obtusifoliol

-

additional information

-

additional information

7.5

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assay at

285347, 285351

30

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UniProt

endoplasmic reticulum

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distribution of Erg proteins between endoplasmic reticulum and liquid particles in wild-type and DELTAerg11-DELTAerg3 mutant, overview

5783

membrane

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16020

microsome

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285344, 285345, 285351, 285354, 672414

additional information

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comparison of CYP51 family enzyme structures, overview

X-ray crystal structures of hexahistidine-tagged Saccharomyces cerevisiae lanosterol 14alpha-demethylase in complex with its substrate lanosterol, the pseudosubstrate estriol and the triazole drugs itraconazole, posaconazole, fluconazole and voriconazole

Y140F

3.3fold reduction in susceptibility to S-prothioconazole

Y140H

4.3fold reduction in susceptibility to S-prothioconazole

additional information

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construction of a deletion mutant strain DELTAerg11, the defect is lethal, but mutants survive in a DELTAerg3 background, erg11/erg3 mutant lacks ergosterol but compensates the lack, sterol distrubution is altered, mutant sterol composition and phenotype, overview

to homogeneity

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CYP51 cDNA cloned with pBluescript KS-. Self sufficient lanosterol 14-demethylase fusion proteins by using yeast CYP51 and P450 reductase, expressed in Escherichia coli

expression of N-terminally GFP-tagged enzyme in strain FYN8 under control of the GAL1 promoter

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recombinant hexahistidine-tagged enzyme is overexpresssed in a yeast membrane protein expression system

sequence comparison, phylogenetic analysis, heterologous overexpression

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agriculture

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target enzyme for azole antifungal agents. These specific inhibitors are of great importance as plant growth regulators, fungicides and herbicides in the agricultural and medical fields

medicine

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target enzyme for azole antifungal agents. These specific inhibitors are of great importance as plant growth regulators, fungicides and herbicides in the agricultural and medical fields

pharmacology

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target enzyme for azole antifungal agents. These specific inhibitors are of great importance as plant growth regulators, fungicides and herbicides in the agricultural and medical fields

285344

Bak, S.; Kahn, R.A.; Olsen, C.E.; Halkier, B.A.

Cloning and expression in Escherichia coli of the obtusifoliol 14alpha-demethylase of Sorghum bicolor (L.) Moench, a cytochrome P450 orthologous to the sterol 14alpha-demethylases (CYP51) from fungi and mammals

Plant J.

11

191-201

1997

Arabidopsis thaliana, Saccharomyces cerevisiae, Candida albicans, [Candida] glabrata, Candida tropicalis, Homo sapiens, Manihot esculenta, no activity in Escherichia coli, Penicillium italicum, Rattus norvegicus, Schizosaccharomyces pombe, Sinapis alba, Sorghum bicolor, Ustilago maydis, Zea mays, Sinapis alba L.

9076987

285345

Aoyama, Y.; Yoshida, Y.

Different substrate specificities of lanosterol 14alpha-demethylase (P-45014DM) of Saccharomyces cerevisiae and rat liver for 24-methylene-24,25-dihydrolanosterol and 24,25-dihydrolanosterol

Biochem. Biophys. Res. Commun.

178

1064-1071

1991

Saccharomyces cerevisiae, Rattus norvegicus

1872829

Aoyama, Y.; Yoshida, Y.

The 4beta-methyl group of substrate does not affect the activity of lanosterol 14alpha-demethylase (P-45014DM) of yeast: Difference between the substrate recognition by yeast and plant sterol 14alpha-demethylases

Biochem. Biophys. Res. Commun.

183

1266-1272

1992

Saccharomyces cerevisiae, Embryophyta

1567403

Aoyama, Y.; Yoshida, Y.; Sonoda, Y.; Sato, Y.

Role of the 8-double bond of lanosterol in the enzyme-substrate interaction of cytochrome P-45014DM (lanosterol 14alpha-demethylase)

Biochim. Biophys. Acta

1001

196-200

1989

Saccharomyces cerevisiae

2917143

285348

Aoyama, Y.; Funae, Y.; Noshiro, M.; Horiuchi, T.; Yoshida, Y.

Occurrence of a P450 showing high homology to yeast lanosterol 14-demethylase (P45014DM) in the rat liver

Biochem. Biophys. Res. Commun.

201

1320-1326

1994

Saccharomyces cerevisiae, Candida albicans, Candida tropicalis, Embryophyta, Rattus norvegicus

8024575

Aoyama, Y.; Yoshida, Y.; Sato, R.

Yeast cytochrome P-450 catalyzing lanosterol 14alpha-demethylation. II. Lanosterol metabolism by purified P-45014DM and by intact microsomes

J. Biol. Chem.

259

1661-1666

1984

Saccharomyces cerevisiae

6420412

285352

Van Nistelrooy, J.G.M.; Van den Brink, J.M.; Van Kan, J.A.L.; Van Gorcom, R.F.M.; de Waard, M.A.

Isolation and molecular characterisation of the gene encoding eburicol 14alpha-demethylase (CYP51) from Penicillium italicum

Mol. Gen. Genet.

250

725-733

1996

Saccharomyces cerevisiae, Candida albicans, Candida tropicalis, Penicillium italicum

8628233

285354

Aoyama, Y.; Noshiro, M.; Gotoh, O.; Imaoka, S.; Funae, Y.; Kurosawa, N.; Horiuchi, T.; Yoshida, Y.

Sterol 14-demethylase P450 (P45014DM*) is one of the most ancient and conserved P450 species

J. Biochem.

119

926-933

1996

Saccharomyces cerevisiae, Candida albicans, Candida tropicalis, Embryophyta, Homo sapiens, Penicillium italicum, Rattus norvegicus, Schizosaccharomyces pombe, Ustilago maydis

8797093

Ott, R.G.; Athenstaedt, K.; Hrastnik, C.; Leitner, E.; Bergler, H.; Daum, G.

Flux of sterol intermediates in a yeast strain deleted of the lanosterol C-14 demethylase Erg11p

Biochim. Biophys. Acta

1735

111-118

2005

Saccharomyces cerevisiae, Saccharomyces cerevisiae FY1679

15951236

Lepesheva, G.I.; Waterman, M.R.

Sterol 14alpha-demethylase cytochrome P 450 (CYP51), a P450 in all biological kingdoms

Biochim. Biophys. Acta

1770

467-477

2007

Candida albicans, Homo sapiens, Methylococcus capsulatus, Mycobacterium tuberculosis, Mycolicibacterium smegmatis, Rattus norvegicus, Saccharomyces cerevisiae, Sorghum bicolor, Trypanosoma brucei, Trypanosoma cruzi, Ustilago maydis

16963187

Parker, J.E.; Merkamm, M.; Manning, N.J.; Pompon, D.; Kelly, S.L.; Kelly, D.E.

Differential azole antifungal efficacies contrasted using a Saccharomyces cerevisiae strain humanized for sterol 14 alpha-demethylase at the homologous locus

Antimicrob. Agents Chemother.

52

3597-3603

2008

Homo sapiens (Q16850), Homo sapiens, Saccharomyces cerevisiae, Saccharomyces cerevisiae BY4741

18694951

Kitahama, Y.; Nakamura, M.; Yoshida, Y.; Aoyama, Y.

The construction and characterization of self-sufficient lanosterol 14-demethylase fusion proteins consisting of yeast CYP51 and its reductase

Biol. Pharm. Bull.

32

558-563

2009

Saccharomyces cerevisiae (P10614), Saccharomyces cerevisiae

19336884