Information on EC 2.1.1.142 - cycloartenol 24-C-methyltransferase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.1.1.142
-
RECOMMENDED NAME
GeneOntology No.
cycloartenol 24-C-methyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
S-adenosyl-L-methionine + cycloartenol = S-adenosyl-L-homocysteine + (24R)-24-methylcycloart-25-en-3beta-ol
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
transfer of methyl group
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
ergosterol biosynthesis II
-
-
SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:cycloartenol 24-C-methyltransferase
S-Adenosyl-L-methionine methylates the Si face of the 24(25)-double bond with elimination of a hydrogen atom from the pro-Z methyl group at C-25.
CAS REGISTRY NUMBER
COMMENTARY hide
37257-07-1
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
gene PbSMT
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
24(28)-methylene lophenol + S-adenosyl-L-methionine
24(28)Z-ethylidene lophenol + S-adenosyl-L-homocysteine
show the reaction diagram
-
-
-
?
24(28)-methylenecholest-7-enol + S-adenosyl-L-methionine
?
show the reaction diagram
-
-
-
-
?
24(28)-methylenecholesterol + S-adenosyl-L-methionine
?
show the reaction diagram
-
-
-
-
?
24(28)-methylenecycloartanol + S-adenosyl-L-methionine
?
show the reaction diagram
-
-
-
-
?
24(28)-methylenelanosterol + S-adenosyl-L-methionine
?
show the reaction diagram
-
-
-
-
?
24,25-dehydro-27-methenyl-cycloartenol + S-adenosyl-L-methionine
?
show the reaction diagram
-
CA-8, i.e. cycloartenol nucleus substrate analogue with double bond at C8, an enzyme inhibitor, is a poor substrate
-
-
?
24,25-dehydro-27-methylene-cycloartenol + S-adenosyl-L-methionine
?
show the reaction diagram
-
CA-14, i.e. cycloartenol nucleus substrate analogue with double bond at C14, an enzyme inhibitor, is a poor substrate
-
-
?
24-fluorocycloartenol + S-adenosyl-L-methionine
(24R)-24-methyl-24-fluoro-cycloart-25-en-3beta-ol + S-adenosyl-L-homocysteine
show the reaction diagram
-
allylic substrate analog
-
-
?
26,27-dehydrozymosterol + S-adenosyl-L-methionine
26-homocholesta-8(9),23(24)E,26(26')-trienol + 26-homocholesta-8(9),26(26')-3beta,24beta-dienol
show the reaction diagram
-
-
-
-
?
31-norcycloartenol + S-adenosyl-L-methionine
?
show the reaction diagram
-
-
-
-
?
4alpha-methylfecosterol + S-adenosyl-L-methionine
?
show the reaction diagram
-
-
-
-
?
4alpha-methylzymosterol + S-adenosyl-L-methionine
?
show the reaction diagram
-
-
-
-
?
cholest-7,24-dienol + S-adenosyl-L-methionine
?
show the reaction diagram
-
-
-
-
?
cycloartenol + S-adenosyl-L-methionine
(24R)-24-methylcycloart-25-en-3beta-ol + S-adenosyl-L-homocysteine
show the reaction diagram
-
-
-
-
?
cycloartenol + S-adenosyl-L-methionine
24(28)-methylenecycloartenol + S-adenosyl-L-homocysteine
show the reaction diagram
-
-
-
-
?
cycloartenol + S-adenosyl-L-methionine
24-methylenecycloartenol + S-adenosyl-L-homocysteine
show the reaction diagram
cyclobranol + S-adenosyl-L-methionine
9beta-25-methylidene-9,19-cyclolanost-1-ene-1,3-diol + S-adenosyl-L-homocysteine
show the reaction diagram
-
-
-
-
?
dehydropollinastanol + S-adenosyl-L-methionine
?
show the reaction diagram
-
-
-
-
?
desmosterol + S-adenosyl-L-methionine
?
show the reaction diagram
ergosta-7,24(28)-dienol + S-adenosyl-L-methionine
? + S-adenosyl-L-homocysteine
show the reaction diagram
fecosterol and ergosta-7,24(28)-dienol which lack C4-and C14-methyl groups in the nucleus are catalyzed equally effectively by SMT2-2 as the natural substrate 24(28)-methylene lophenol
-
-
?
fecosterol + S-adenosyl-L-methionine
?
show the reaction diagram
-
-
-
-
?
fecosterol + S-adenosyl-L-methionine
? + S-adenosyl-L-homocysteine
show the reaction diagram
fecosterol and ergosta-7,24(28)-dienol which lack C4-and C14-methyl groups in the nucleus are catalyzed equally effectively by SMT2-2 as the natural substrate 24(28)-methylene lophenol
-
-
?
lanosterol + S-adenosyl-L-methionine
? + S-adenosyl-L-homocysteine
show the reaction diagram
-
-
-
?
obtusifoliol + S-adenosyl-L-methionine
?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + 14-methylzymosterol
?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + 14alpha-methylzymosterol
S-adenosyl-L-homocysteine + ?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + 24(28)-methylenecycloartanol
S-adenosyl-L-homocysteine + ?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + 24(28)-methylenelanosterol
S-adenosyl-L-homocysteine + ?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + 24(28)-methylenelophenol
S-adenosyl-L-homocysteine + ?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + 24-dehydropollinstanol
S-adenosyl-L-homocysteine + ?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + 24-fluorocycloartenol
S-adenosyl-L-homocysteine + ?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + 24-methyl-9,19-cyclolanost-24-en-3-ol
S-adenosyl-L-homocysteine + 25-methyl-24-methylidene-9,19-cyclolanostan-3-ol
show the reaction diagram
-
i.e. cyclobranol
-
-
?
S-adenosyl-L-methionine + 26,27-dehydrocycloartenol
S-adenosyl-L-homocysteine + ?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + 26,27-dehydrolanosterol
?
show the reaction diagram
-
Formation of a reversibly bound enzyme–substrate complex, followed by catalysis to an intermediate that can be converted to a methyl product or the intermediate can be intercepted through covalent modification and hence irreversible inhibition, lanosterol or 26,27-dehydrolanosterol can lead to distinct intermediates that convert to methylated sterol products
-
-
?
S-adenosyl-L-methionine + 26-difluorocycloartenol
?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + 26-fluorocycloartenol
?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + 30,31-dinorcycloartenol
S-adenosyl-L-homocysteine + ?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + 31-norcycloartenol
S-adenosyl-L-homocysteine + ?
show the reaction diagram
S-adenosyl-L-methionine + 31-norlanosterol
?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + cholesta-5,20(22)E,24-trienol
S-adenosyl-L-homocysteine + ?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + cholesta-5,7,22E,24-tetraenol
S-adenosyl-L-homocysteine + ?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + cycloartenol
S-adenosyl-L-homocysteine + (24R)-24-methylcycloart-25-en-3beta-ol
show the reaction diagram
S-adenosyl-L-methionine + cycloartenol
S-adenosyl-L-homocysteine + 24(28)-methylenecycloartanol
show the reaction diagram
S-adenosyl-L-methionine + cycloartenol
S-adenosyl-L-homocysteine + 24(28)methylene cycloartanol
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + cycloartenol
S-adenosyl-L-homocysteine + 24-methylene cycloartenol
show the reaction diagram
S-adenosyl-L-methionine + cycloartenol
S-adenosyl-L-homocysteine + 24-methylenecycloartenol
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + cycloartenol
S-adenosyl-L-homocysteine + ?
show the reaction diagram
S-adenosyl-L-methionine + lanosterol
?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + lanosterol
S-adenosyl-L-homocysteine + 24(28)-methylene-24,25-dihydro-lanosterol
show the reaction diagram
S-adenosyl-L-methionine + lanosterol
S-adenosyl-L-homocysteine + ?
show the reaction diagram
S-adenosyl-L-methionine + parkeol
S-adenosyl-L-homocysteine + ?
show the reaction diagram
S-adenosyl-L-methionine + zymosterol
S-adenosyl-L-homocysteine + ?
show the reaction diagram
zymosterol + S-adenosyl-L-methionine
fecosterol + S-adenosyl-L-homocysteine
show the reaction diagram
24(28)-methylenelophenol + S-adenosyl-L-methionine
additional information
-
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
cycloartenol + S-adenosyl-L-methionine
(24R)-24-methylcycloart-25-en-3beta-ol + S-adenosyl-L-homocysteine
show the reaction diagram
-
-
-
-
?
cycloartenol + S-adenosyl-L-methionine
24-methylenecycloartenol + S-adenosyl-L-homocysteine
show the reaction diagram
-
SMT2 has a position-specific substrate specificity for DELTA24(25)-sterols and contains a single active center to catalyze the consecutive C1-transfer activities by substrate reaction channels similar to the fungal SMT1
-
-
?
S-adenosyl-L-methionine + cycloartenol
S-adenosyl-L-homocysteine + (24R)-24-methylcycloart-25-en-3beta-ol
show the reaction diagram
S-adenosyl-L-methionine + cycloartenol
S-adenosyl-L-homocysteine + 24(28)-methylenecycloartanol
show the reaction diagram
S-adenosyl-L-methionine + cycloartenol
S-adenosyl-L-homocysteine + 24-methylene cycloartenol
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + lanosterol
S-adenosyl-L-homocysteine + 24(28)-methylene-24,25-dihydro-lanosterol
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
S-adenosyl-L-methionine
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(3beta24R,S)-24-methyl-24-thionacycloart-3-ol iodide
-
reversible, non-competitive-type
24(28)-methylene cycloartanol
; competitive
24(28)-methylenecycloartanol
24(28)-methylenencycloartanol
-
-
24,25-dehydro-27-methenyl-cycloartenol
-
i.e. cycloartenol nucleus substrate analogue with double bond at C8
24,25-dehydro-27-methylene-cycloartenol
-
i.e. cycloartenol nucleus substrate analogue with double bond at C14, non-competitive
24-bromocycloartenol
-
competitive; competitive inhibitor relative to cycloartenol; time-dependent inhibition kinetics
24-dehydrocycloartenol
-
a mechanism-based inactivator
24-fluorocycloartenol
-
allylic substrate analog, time-dependent inhibition kinetics, consistent with the action of a suicide substrate that irreversibly inactivates the enzyme; competitive, the electron-withdrawing alpha-fluorine substituent suppresses the rate of the C-methylation reaction
24-methyl-9,19-cyclolanost-24-en-3-ol
-
i.e. cyclobranol, competitive
24-thiacycloartanol
24-thiacycloartenol
-
-
24beta-methylcycloartanol
-
competitive with cycloartenol
25-azacycloartanol
; non-competitive
25-Azacycloartenol
25-azalanosterol
-
substrate analogue, competitive-type inhibitor, a potent inhibitor of cell growth promoting lanosterol accumulation and 24-alkyl sterol depletion
26,27-dehydrocycloartenol
-
-
26,27-dehydrolanosterol
-
substrate analogue, competitive-type inhibitor, pseudofirst-order, time-dependent inactivation of the PbSMT. Formation of a reversibly bound enzyme-substrate complex, followed by catalysis to an intermediate that can be converted to a methyl product or the intermediate can be intercepted through covalent modification and hence irreversible inhibition, lanosterol or 26,27-dehydrolanosterol can lead to distinct intermediates that convert to methylated sterol products
26,27-dehydrozymosterol
-
mechanism-based inhibitor, specific covalent modification of SMT2
26-difluorocycloartenol
-
competitive inhibitor
-
26-fluorocycloartenol
-
-
-
26-methenylcycloartenol
-
pseudo-first-order time-dependent inactivation, less potency of inhibition
26-methynylcycloartenol
-
inhibition kinetics non-competitive and time-dependent, less potency of inhibition
citrostadienol
-
competitive with cycloartenol or 24(28)-methylenelophenol
cycloarta-24,26-dienol
-
-
Cycloartenol
-
-
cyclobranol
-
competitive-type inhibitor
cyclolaudenol
-
dead end inhibitor analog, competitive versus cycloartenol, uncompetitive versus S-adenosyl-L-methionine
S-adenosyl-L-homocysteine
-
noncompetitive with respect to S-adenosyl-L-methionine
sitosterol
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.125
14alpha-Methylzymosterol
-
-
0.035
24(28)-methylenecholest-7-enol
-
pH 7.5, 35°C
0.036
24(28)-methylenecholesterol
-
pH 7.5, 35°C
0.022 - 0.04
24(28)-methylenecycloartanol
0.02 - 0.035
24(28)-methylenelanosterol
0.022 - 0.03
24(28)-methylenelophenol
0.028
24(28)methylene lophenol
-
0.005
24,25-dehydro-27-methylene-cycloartenol
-
pH 7.5, 35°C, recombinant enzyme
0.045
24-dehydropollinstanol
-
-
0.03 - 0.033
24-fluorocycloartenol
0.037
24-methyl-9,19-cyclolanost-24-en-3-ol
-
-
0.01
26,27-dehydrocycloartenol
-
-
0.015
26,27-dehydrozymosterol
-
pH 7.5, 35°C
0.044
26-difluorocycloartenol
-
at 35°C in phosphate buffer at pH 8.0
-
0.036
26-fluorocycloartenol
-
at 35°C in phosphate buffer at pH 8.0
-
0.97
30,31-dinorcycloartenol
-
-
-
0.03 - 0.61
31-Norcycloartenol
-
0.03
4alpha-methylfecosterol
-
pH 7.5, 35°C
0.03
4alpha-methylzymosterol
-
pH 7.5, 35°C
0.03
cholest-7,24-dienol
-
pH 7.5, 35°C
0.1 - 0.276
cholesta-5,20(22)E,24-trienol
0.112
cholesta-5,7,22E,24-tetraenol
-
-
0.024 - 0.045
Cycloartenol
0.087
cyclobranol
-
-
0.023
dehydropollinastanol
-
pH 7.5, 35°C
0.038 - 0.063
desmosterol
0.016
fecosterol
-
pH 7.5, 35°C
0.027 - 0.176
lanosterol
0.038
obtusifoliol
-
pH 7.5, 35°C
0.043 - 0.8
parkeol
0.028 - 0.038
Zymosterol
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.001
24(28)-methylenelophenol
Glycine max
-
-
0.013
24(28)methylene lophenol
Glycine max
D2D5G4
-
0.00000083
24,25-dehydro-27-methylene-cycloartenol
Glycine max
-
pH 7.5, 35°C, recombinant enzyme
0.0003 - 0.00033
24-fluorocycloartenol
0.00033
26,27-dehydrocycloartenol
Glycine max
-
-
0.001
26,27-dehydrozymosterol
Arabidopsis thaliana
-
pH 7.5, 35°C
0.01
Cycloartenol
0.0001
cyclobranol
Glycine max
-
-
0.0023
lanosterol
Paracoccidioides brasiliensis
-
pH 7.5, 30°C, recombinant wild-type enzyme
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000055
(3beta24R,S)-24-methyl-24-thionacycloart-3-ol iodide
-
-
0.009 - 0.25
24(28)-methylene cycloartanol
0.171
24(28)-methylenencycloartanol
-
-
0.047
24,25-dehydro-27-methenyl-cycloartenol
-
pH 7.5, 35°C, recombinant enzyme
0.067
24,25-dehydro-27-methylene-cycloartenol
-
pH 7.5, 35°C, recombinant enzyme
0.036
24-bromocycloartenol
-
; competitive inhibition, concentration of substrate analogs varied at 5, 15, 25, and 50 microM; pH 7.5, 35°C, versus cycloartenol
0.042
24-dehydrocycloartenol
-
pH 7.5, 35°C, recombinant enzyme
0.032
24-fluorocycloartenol
-
competitive inhibition, concentration of substrate analogs varied at 5, 15, 25, and 50 microM; pH 7.5, 35°C, versus cycloartenol
0.025
24-methyl-9,19-cyclolanost-24-en-3-ol
-
-
0.002
24-thiacycloartanol
0.15
24beta-methylcycloartanol
-
pH 7.4, 35°C, substrate cycloartenol
0.00001 - 0.00002
25-azacycloartanol
0.00002 - 0.00004
25-Azacycloartenol
0.000014
25-azalanosterol
-
pH 7.5, 30°C, recombinant wild-type enzyme
0.042
26,27-dehydrocycloartenol
-
-
0.054
26,27-dehydrolanosterol
-
pH 7.5, 30°C, recombinant wild-type enzyme
0.049
26,27-dehydrozymosterol
-
pH 7.5, 35°C
0.093
26-difluorocycloartenol
-
at 35°C in phosphate buffer at pH 8.0
-
0.071
26-fluorocycloartenol
-
at 35°C in phosphate buffer at pH 8.0
-
0.047
26-methenylcycloartenol
-
rate of inactivation saturable, comparable to values obtained for cycloartenol
0.045 - 0.05
citrostadienol
0.017
cycloarta-24,26-dienol
-
-
0.03
Cycloartenol
-
natural substrate
0.025
cyclobranol
-
-
0.007 - 0.009
cyclolaudenol
0.047 - 0.056
S-adenosyl-L-homocysteine
0.1 - 0.207
sitosterol
additional information
additional information
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00003
25-azalanosterol
Paracoccidioides brasiliensis
-
pH 7.5, 30°C, recombinant wild-type enzyme
0.067
26-methynylcycloartenol
Glycine max
-
co-incubation with 50 microM or 100 microM cycloartenol affords protection against inactivation generating 25% and 45% C-methylation
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.8 - 8.9
-
-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 9
-
half-maximal activities at pH 6.5 and pH 9.0
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35
-
assay at; inhibition assay at
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.3
calculated
6.5
calculated
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
young leaf; young leaf
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40340
4 * 40340, calculated, 4 * 40000, SDS-PAGE
40397
x * 40397, calculated
41500
-
4 * 41500, SDS-PAGE
42502
-
x * 42502, sequence calculation
43000
-
SDS-PAGE
154000
-
gel filtration
160000
161000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
tetramer
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
partially purified, recombinant protein, SDS-PAGE; recombinant enzyme from Escherichia coli to homogeneity
-
partially purified; SDS-PAGE
-
Q-Sepharose column chromatography
-
recombinant enzyme
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
-
expressed in Escherichia coli, recombinant protein; expression in Escherichia coli
-
expressed in Escherichia coli; expressed in Escherichia coli, critical 24H replaced by a fluorine atome
-
expression in Escherichia coli
-
expression in Escherichia coli; expression in Escherichia coli
gene PbSMT, DNA and amino acid sequence determination and analysis, phylogenetic analysis, overexpression of soluble His-tagged wild-type and mutant enzymes in Escherichia coli strains BL21(DE3) and BL21(C43)
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Y83F
-
Km and kcat, of wild-type enzyme and mutant enzyme are similar for cycloartenol as substrate
Y83L
-
Km and kcat, of wild-type enzyme and mutant enzyme are similar for cycloartenol as substrate
Y81F
-
site-directed mutagenesis, the mutation causes a preferential change in affinity for DELTA24-sterols that affords alter partitioning in the direction of 24-ethyl(idene) product formation
Y81L
-
site-directed mutagenesis, the mutation causes a preferential change in affinity for DELTA24-sterols that affords alter partitioning in the direction of 24-ethyl(idene) product formation
Y88F
-
site-directed mutagenesis, the mutation causes a 10% loss in activity compared to the wild-type enzyme
Y88L
-
site-directed mutagenesis, the mutation causes a 50% loss in activity compared to the wild-type enzyme
Y81F
-
site-directed mutagenesis, the mutation causes a preferential change in affinity for DELTA24-sterols that affords alter partitioning in the direction of 24-ethyl(idene) product formation
-
Y81L
-
site-directed mutagenesis, the mutation causes a preferential change in affinity for DELTA24-sterols that affords alter partitioning in the direction of 24-ethyl(idene) product formation
-
Y88F
-
site-directed mutagenesis, the mutation causes a 10% loss in activity compared to the wild-type enzyme
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Y88L
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site-directed mutagenesis, the mutation causes a 50% loss in activity compared to the wild-type enzyme
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