Information on EC 1.14.13.72 - methylsterol monooxygenase

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The enzyme appears in viruses and cellular organisms

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EC NUMBER
COMMENTARY hide
1.14.13.72
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RECOMMENDED NAME
GeneOntology No.
methylsterol monooxygenase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carbaldehyde + NAD(P)H + H+ + O2 = 3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carboxylate + NAD(P)+ + H2O
show the reaction diagram
4,4-dimethyl-5alpha-cholest-7-en-3beta-ol + NAD(P)H + H+ + O2 = 4beta-hydroxymethyl-4alpha-methyl-5alpha-cholest-7-en-3beta-ol + NAD(P)+ + H2O
show the reaction diagram
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-
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4beta-hydroxymethyl-4alpha-methyl-5alpha-cholest-7-en-3beta-ol + NAD(P)H + H+ + O2 = 3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carbaldehyde + NAD(P)+ + 2 H2O
show the reaction diagram
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-
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-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
demethylation
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oxidation
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-
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redox reaction
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-
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reduction
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of antibiotics
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Biosynthesis of secondary metabolites
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Metabolic pathways
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Steroid biosynthesis
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cholesterol biosynthesis
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SYSTEMATIC NAME
IUBMB Comments
4,4-dimethyl-5alpha-cholest-7-en-3beta-ol,NAD(P)H:oxygen oxidoreductase (hydroxylating)
Requires cytochrome b5. Also acts on 4alpha-methyl-5alpha-cholest-7-en-3beta-ol. The sterol can be based on cycloartenol as well as lanosterol.
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CAS REGISTRY NUMBER
COMMENTARY hide
37256-80-7
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Sprague-Dawley
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-
Manually annotated by BRENDA team
endomycorrhizal fungus, grown on chicory (Cichorium intybus) roots
UniProt
Manually annotated by BRENDA team
strain FL530, enzyme is a component of the sterol-4-demethylation multienzyme complex
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Manually annotated by BRENDA team
SMO1
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Manually annotated by BRENDA team
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
first of three steps to remove 2 C4 methyl groups from an intermediate in ergosterol biosynthesis in the sterol biosynthetic pathway, an enzymatic complex is formed with other enzymes in the pathway
physiological function
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the enzyme regulates epidermal growth factor receptor signaling and vesicular trafficking
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
24-ethylidene-24-dihydrolophenol + NADPH + O2
?
show the reaction diagram
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isozyme SMO2
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-
?
24-ethylidenelophenol + NAD(P)H + O2
avenasterol + NAD(P)+ + H2O
show the reaction diagram
-
best substrate, 4alpha-carbomethoxy-5alpha-stigmasta-7,24(241)-dien-3beta-yl-acteate and avenastenone as intermediates
-
?
24-methylene-24-dihydrocycloartenol + NADPH + O2
?
show the reaction diagram
24-methylenecycloartanol + NAD(P)H + O2
cycloeucalenol + NAD(P)+ + H2O
show the reaction diagram
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best substrate, 4beta,14alpha-dimethyl-4alpha-acetoxymethyl-5alpha-ergosta-9beta,19-cyclo-24(241)-en-3beta-yl-acetate, 4beta-,14alpha-dimethyl-4alpha-carbomethoxy-5alpha-ergosta-9beta,19-cyclo-24(241)-en-3beta-yl-acetate and cycloeucalenone as intermediates
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?
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carbaldehyde + NAD(P)H + H+ + O2
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carboxylate + NAD(P)+ + H2O
show the reaction diagram
1c
-
-
?
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carbaldehyde + NAD(P)H + O2
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carboxylate + NAD(P)+ + H2O
show the reaction diagram
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carbaldehyde + NADPH + O2
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carboxylate + NADP+ + H2O
show the reaction diagram
4,4-dimethyl-5alpha-cholest-7-en-3beta-ol + 3 NAD(P)H + 3 H+ + 3 O2
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carboxylate + 3 NAD(P)+ + 4 H2O
show the reaction diagram
overall reaction
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-
?
4,4-dimethyl-5alpha-cholest-7-en-3beta-ol + NAD(P)H + H+ + O2
4alpha-hydroxymethyl-4beta-methyl-5alpha-cholest-7-en-3beta-ol + NAD(P)+ + H2O
show the reaction diagram
1a
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-
?
4,4-dimethyl-5alpha-cholest-7-en-3beta-ol + NAD(P)H + O2
4alpha-methyl-5alpha-cholest-7-en-3beta-ol + NAD(P)+ + CO2
show the reaction diagram
4,4-dimethyl-5alpha-cholest-7-en-3beta-ol + NAD(P)H + O2
4beta-hydroxymethyl-4alpha-methyl-5alpha-cholest-7-en-3beta-ol + NAD(P)+ + H2O
show the reaction diagram
4,4-dimethyl-5alpha-cholest-7-en-3beta-ol + NADPH + O2
4beta-hydroxymethyl-4alpha-methyl-5alpha-cholest-7-en-3beta-ol + NADP+ + H2O
show the reaction diagram
4,4-dimethyl-5alpha-cholesta-8,14,24-trien-3beta-ol + 3 NAD(P)H + 3 O2
(3beta,4alpha)-3-hydroxy-4-methylcholesta-8,14-24-triene-4-carboxylate + 3 NAD(P)+ + 3 H2O
show the reaction diagram
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-
-
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?
4,4-dimethyl-zymosterol + NADPH + O2
4-hydroxymethyl,4-methyl-zymosterol + NADP+ + H2O
show the reaction diagram
4,4-dimethyl-zymosterol + NADPH + O2
?
show the reaction diagram
4alpha-hydroxymethyl-4beta-methyl-5alpha-cholest-7-en-3beta-ol + NAD(P)H + H+ + O2
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carbaldehyde + NAD(P)+ + 2 H2O
show the reaction diagram
1b
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-
?
4beta-hydroxymethyl-4alpha-methyl-5alpha-cholest-7-en-3beta-ol + NAD(P)H + O2
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carbaldehyde + NAD(P)+ + H2O
show the reaction diagram
4beta-hydroxymethyl-4alpha-methyl-5alpha-cholest-7-en-3beta-ol + NADPH + O2
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carbaldehyde + NADP+ + H2O
show the reaction diagram
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
24-methylene-24-dihydrocycloartenol + NADPH + O2
?
show the reaction diagram
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-
?
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carbaldehyde + NAD(P)H + O2
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carboxylate + NAD(P)+ + H2O
show the reaction diagram
-
step 3 of 3 of the overall reaction
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?
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carbaldehyde + NADPH + O2
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carboxylate + NADP+ + H2O
show the reaction diagram
4,4-dimethyl-5alpha-cholest-7-en-3beta-ol + NAD(P)H + O2
4beta-hydroxymethyl-4alpha-methyl-5alpha-cholest-7-en-3beta-ol + NAD(P)+ + H2O
show the reaction diagram
-
step 1 of 3 of the overall reaction
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?
4,4-dimethyl-5alpha-cholest-7-en-3beta-ol + NADPH + O2
4beta-hydroxymethyl-4alpha-methyl-5alpha-cholest-7-en-3beta-ol + NADP+ + H2O
show the reaction diagram
4,4-dimethyl-zymosterol + NADPH + O2
4-hydroxymethyl,4-methyl-zymosterol + NADP+ + H2O
show the reaction diagram
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a step in the ergosterol biosynthesis, pathway, overview
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?
4beta-hydroxymethyl-4alpha-methyl-5alpha-cholest-7-en-3beta-ol + NAD(P)H + O2
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carbaldehyde + NAD(P)+ + H2O
show the reaction diagram
-
step 2 of 3 of the overall reaction
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?
4beta-hydroxymethyl-4alpha-methyl-5alpha-cholest-7-en-3beta-ol + NADPH + O2
3beta-hydroxy-4beta-methyl-5alpha-cholest-7-ene-4alpha-carbaldehyde + NADP+ + H2O
show the reaction diagram
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cytochrome b5
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dependent on
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NAD(P)H
NADPH
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe2+
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presumably, or other metal ion suggested to be enzyme-boud in hydroxylating system
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6-amino-2-n-pentylbenzothiazole
cyanide
cytochrome c
fenhexamid
fungicide, sterol biosynthesis inhibitor, 9.3 mg/l growth medium (long term exposure for 8 weeks) inhibits germ tube length by 50%, no change in enzyme transcription (0.2-2 mg/l growth medium)
fenpropimorph
fungicide, sterol biosynthesis inhibitor, 6.4 mg/l growth medium (long term exposure for 8 weeks) inhibits germ tube length by 50%, strong decrease in transcription levels with 2 mg/l growth medium, 10 mg/l decreases the fraction of vital stained fungal cells (after 2 and 7 days of treatment), leads to a increase in enzyme mRNA, does not change relative sterol composition and sterol amounts but reduced squalen content by 87% after 7 days of treatment
IgG
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polyclonal antibody raised against recombinant cytochrome b5, strongly inhibited of enzyme activity by low amounts of IgG, this results from IgG specific to cytochrome b5
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metal chelating agent
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o-phenanthroline
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stigmastan-3beta,5alpha,6alpha-triol
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metabolism of 24-ethylidenelophenol effectively inhibited. Metabolism of 24-methylenecycloartanol unaffected
Trypsin
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30 min treatment, 18% of original concentration of cytochrome b5 left
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additional information
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.48
24-ethylidenelophenol
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+/-0.02
0.15
24-methylcycloartanol
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+/-0.01
0.34
24-methylenelophenol
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+/-0.02
0.09 - 0.5
cycloeucalenol
0.25
cyclolaudenol
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+/-0.01
additional information
additional information
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.000201
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recovered after Lubrol treatment, 1% of original cytochrome b5 concentration in microsomes, 4-fold that observed for microsomes, maybe cytochrome b5 does not participate in hydroxylation
additional information
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5 - 8.5
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-
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
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assay at
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pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.8
Pcerg25B, sequence calculation
7.36
Pcerg25A, sequence calculation
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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epidermal
Manually annotated by BRENDA team
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
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the enzyme forms a demethylation complex in the ER together with the transmembrane protein Erg28p, and possibly the other C-4 demethylation proteins Erg26p, overview
Manually annotated by BRENDA team
additional information
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subcellular localization study
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Manually annotated by BRENDA team
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35000
calculated from amino acids
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SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 34 500, Pcerg25B, sequence calculation; x * 34 873, Pcerg25A, sequence calculation
additional information
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
remarkable stability during 4 to 6 h incubation periods
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
microsomes frozen, buffers contain 1 mM glutathione, no loss of activity
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
extraradial mycelium collected from solubilized medium in buffer (50 mM Tris-HCl, pH 7.5, 10 mM EDTA), vacuum filtrated, frozen in liquid nitrogen
gel filtration, ammonium sulfate fractionation, affinity chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis
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DNA and amino acid sequence determination and analysis of wild-type and mutant strains
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DNA and amino acid sequence determination and analysis, expression in enzyme-lacking Saccharomyces cerevisiae strain erg25; DNA and amino acid sequence determination and analysis, expression in enzyme-lacking Saccharomyces cerevisiae strain erg25; DNA and amino acid sequence determination and analysis, expression in enzyme-lacking Saccharomyces cerevisiae strain erg25; DNA and amino acid sequence determination and analysis, expression in enzyme-lacking Saccharomyces cerevisiae strain erg25; DNA and amino acid sequence determination and analysis, expression in enzyme-lacking Saccharomyces cerevisiae strain erg25
DNA and amino acid sequence determination and analysis, in vitro transcription; DNA and amino acid sequence determination and analysis, in vitro transcription
DNA and amino acid sequence determination and analysis; DNA and amino acid sequence determination and analysis
DNA is extracted from extraradial mycelium growing around Cichorium intybus, PCR-amplification, cloned into pGEM-T Vector System for sequencing, for complementation assay in Saccharomyces cerevisiae mutant erg25-25c (methyl oxidase deficient) the GintSMO gene is amplified and cloned into the NotI-linearized yeast expression vector pFL61
gene Pcerg25A, DNA and amino acid sequence determination and analysis, Pcerg25A is located in the DNA region containing the penicillin gene cluster, and thus its copy number is dependent on the patterns of the cluster region, up to eight copies of Pcerg25A are found in the high-productivity strain NCPC 10086, functional complementation of the erg25-deficient Saccharomyces cerevisiae strain ATCC 4024690, subcloning in Escherichia coli strain DH5alpha, phylogenetic analysis, overview; gene Pcerg25B, DNA and amino acid sequence determination and analysis, Pcerg25B is present in just a single copy in Penicillium chrysogenum genomes, functional complementation of the erg25-deficient Saccharomyces cerevisiae strain ATCC 4024690, subcloning in Escherichia coli strain DH5alpha, phylogenetic analysis, overview
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
N48D/V133A/F135S
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construction of diverse mutants by random mutagenesis, isolation of the temperature-sensitive mutant strain mERG25 which grows normally at 28°C, but fails to grow at 38°C showing an altered sterol profile
additional information
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
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enzyme is an antifungal target
medicine
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enzyme is an antifungal target
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LINKS TO OTHER DATABASES (specific for EC-Number 1.14.13.72)
ExplorEnz
ExPASy
KEGG
MetaCyc
SABIO-RK
NCBI: PubMed, Protein, Nucleotide, Structure, Genome, OMIM
IUBMB Enzyme Nomenclature
PROSITE Database of protein families and domains
SYSTERS
Protein Mutant Database
InterPro (database of protein families, domains and functional sites)