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H232S
-
site-directed mutagenesis, mutant energy profile compared to the wild-type enzyme
H232T
-
site-directed mutagenesis, mutant energy profile compared to the wild-type enzyme
N478H/V482I
-
change of amino acids crucial for specificity to the cycloartenol synthase type. Mutant produces 4% lanosterol, 83% parkeol, and 13% cycloartenol from substrate (S)-2,3-epoxysqualene
D139N/G189A/Q481R
induces cataracts in rats, produces only 39% of the lanosterol produced by wild-type
H469-C472del
deletion allele, inactive
C457D
-
less active than the wild-type
C457D/A525C
-
similar activity as the wild-type
C457D/E526A
-
decrease in enzyme functionality
C457D/E526C
-
very sensitive to the thiol-reacting agent dodecylmaleimide, specific activity and thermal stability are severely reduced
C457D/E526D
-
no effect on catalytic avtivity
C457D/E526Q
-
no effect on catalytic avtivity
C457G
-
site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme, overview
C457G/T509G
-
site-directed mutagenesis, the mutations disrupt the pre-existing H-bond to the protonating Asp456 and the intrinsic His234-Tyr510 H-bond network, respectively, and generates achilleol A as the major product
C703D
-
site-directed mutagenesis, the mutant shows unaltered product spectrum compared to the wild-type enzyme
C703G
-
site-directed mutagenesis, the mutant shows altered product spectrum compared to the wild-type enzyme, but not as diverse as mutants C703I and C703H
C703H
-
site-directed mutagenesis, the mutant generates an unusual truncated bicyclic rearranged intermediate, (8R,9R,10R)-polypoda-5,13E,17E,21-tetraen-3beta-ol, related to iridal-skeleton triterpenoid. Numerous oxidosqualene-cyclized truncated intermediates, including tricyclic, unrearranged tetracyclic with 17alpha/beta exocyclic hydrocarbon side chain, rearranged tetracyclic, and chair-chair-chair tricyclic intermediates, are also isolated from the mutant
C703I
-
site-directed mutagenesis, the mutant generates an unusual truncated bicyclic rearranged intermediate, (8R,9R,10R)-polypoda-5,13E,17E,21-tetraen-3beta-ol, related to iridal-skeleton triterpenoid. Numerous oxidosqualene-cyclized truncated intermediates, including tricyclic, unrearranged tetracyclic with 17alpha/beta exocyclic hydrocarbon side chain, rearranged tetracyclic, and chair-chair-chair tricyclic intermediates, are also isolated from the mutant
C703N
-
site-directed mutagenesis, the mutant shows unaltered product spectrum compared to the wild-type enzyme
C703S
-
site-directed mutagenesis, the mutant shows altered product spectrum compared to the wild-type enzyme, but not as diverse as mutants C703I and C703H
C703T
-
site-directed mutagenesis, the mutant shows altered product spectrum compared to the wild-type enzyme, but not as diverse as mutants C703I and C703H
C703V
-
site-directed mutagenesis, the mutant shows altered product spectrum compared to the wild-type enzyme, but not as diverse as mutants C703I and C703H
F445C
-
produces 10% (13alphaH)-iso-malabarica-14(16)-17E,21-trien-3beta-ol, 69% lanosterol, 13% parkeol, 8% 9beta-lanosta-7,24-dien-3beta-ol from substrate (S)-2,3-epoxysqualene
F445D
-
produces 21% (13alphaH)-iso-malabarica-14(16)-17E,21-trien-3beta-ol, 63% lanosterol, 11% parkeol, 5% 9beta-lanosta-7,24-dien-3beta-ol from substrate (S)-2,3-epoxysqualene
F445M
-
produces 7% (13alphaH)-iso-malabarica-14(16)-17E,21-trien-3beta-ol, 65% lanosterol, 18% parkeol, 10% 9beta-lanosta-7,24-dien-3beta-ol from substrate (S)-2,3-epoxysqualene
F445N
-
produces 10% (13alphaH)-iso-malabarica-14(16)-17E,21-trien-3beta-ol, 63% lanosterol, 9% parkeol, 18% 9beta-lanosta-7,24-dien-3beta-ol from substrate (S)-2,3-epoxysqualene
F445T
-
produces 49% (13alphaH)-iso-malabarica-14(16)-17E,21-trien-3beta-ol, 46% lanosterol, 5% 9beta-lanosta-7,24-dien-3beta-ol from substrate (S)-2,3-epoxysqualene
F699A/C703I
-
site-directed mutagenesis, inactive mutant
F699C/I705F
-
the mutant is inactive
F699H
-
the mutant produces lanosterol (13%), protosta-13(17)-dien-3beta-ol (70%), and (17Z)-protosta-17(20),24-dien-3beta-ol (17%)
F699I
-
the mutant produces lanosterol (100%) as the wild type enzyme
F699L
-
the mutant produces lanosterol (100%) as the wild type enzyme
F699M/C703I
-
site-directed mutagenesis, inactive mutant
F699M/I705F
-
the mutant produces 53% lanosterol, 17% (13alphaH)-isomalabarica-14E,17E,21-dien-3beta-ol, 12% (13alphaH)-isomalabarica-14Z,17E,21-dien-3beta-ol, 15% protosta-13(17),24-dien-3beta-ol, and 3% 17alpha-protosta-20(22),24-dien-3beta-ol from (3S)-2,3-oxidosqualene
F699N
-
nonviable mutant, does not produce lanosterol, but protosta-13(17)-dien-3beta-ol (55%), malabarica-14E,17E,21-trien-3beta-ol (5%), 17alpha-protosta-20,24-dien-3beta-ol (24%), and (17Z)-protosta-17(20),24-dien-3beta-ol (16%)
F699P
-
the mutant produces lanosterol (100%) as the wild type enzyme
F699T/C703I
-
site-directed mutagenesis, different oxidosqualene-cyclized truncated intermediates, including tricyclic, unrearranged tetracyclic with 17alpha/beta exocyclic hydrocarbon side chain, rearranged tetracyclic, and chair-chair-chair tricyclic intermediates, are isolated from the mutant
F699T/I105F
-
the mutant produces 36% lanosterol, 45% protosta-13(17),24-dien-3beta-ol, 7% 17alpha-protosta-20,24-dien-3beta-ol, and 12% 17alpha-protosta-20(22),24-dien-3beta-ol from (3S)-2,3-oxidosqualene
F699X
-
TKW14[pERG7F699X] site-saturated mutants allow for ergosterol-independent growth, with the exception of Leu, Ile, His, Met, Pro, and Thr substitutions
H234A
-
products are 17% protosta-12,24-dien-3beta-ol, 13% protosta-20,24-dien-3beta-ol, 30% lanosterol, 40% parkeol
H234C
-
products are 7% protosta-12,24-dien-3beta-ol, 4% protosta-20,24-dien-3beta-ol, 67% lanosterol, 22% parkeol
H234D
-
products are 58% iso-malabarica-14(16),17,21-trien-3beta-ol, 30% lanosterol, 12% parkeol
H234E
-
products are 49% lanosterol, 51% parkeol
H234F
-
products are 66% achilleol A, 14% lanosterol, 20% parkeol
H234G
-
products are 29% protosta-12,24-dien-3beta-ol, 7% protosta-20,24-dien-3beta-ol, 17% lanosterol, 47% parkeol
H234I
-
products are 70% lanosterol, 30% parkeol
H234K
-
no catalytic activity
H234L
-
products are 39% lanosterol, 31% parkeol, 30% iso-malabarica-14(16),17,21-trien-3beta-ol
H234M
-
products are 17% achilleol A, 10% iso-malabarica-14(16),17,21-trien-3beta-ol, 30% lanosterol, 40% parkeol
H234N
-
products are 23% protosta-12,24-dien-3beta-ol, 14% protosta-20,24-dien-3beta-ol, 27% lanosterol, 10% parkeol, 26% iso-malabarica-14(16),17,21-trien-3beta-ol
H234P
-
products are 64% lanosterol, 36% parkeol
H234Q
-
product is 100% lanosterol, like wild-type
H234R
-
no catalytic activity
H234S
-
product is 100% protosta-12,24-dien-3beta-ol
H234T
-
product is 100% parkeol
H234V
-
products are 42% lanosterol, 58% parkeol
H234W
-
product is 100% parkeol
H234W/Y510V
-
the mutant produces achilleol A, (13H)-isomalabarica-14(26),17E,21-trien-3beta-ol, and lanosterol at a 2:8:90 ratio from (S)-2,3-oxidosqualene
H234Y
-
products are 14% achilleol A, 26% protosta-12,24-dien-3beta-ol, 51% lanosterol, 9% parkeol
H234Y/Y510A
-
the mutant produces lanosterol from (S)-2,3-oxidosqualene (wild type reaction)
I705A
-
the mutant produces 78% lanosterol, 13% (13alphaH)-isomalabarica-14E,17E,21-dien-3beta-ol, 8% (13alphaH)-isomalabarica-14Z,17E,21-dien-3beta-ol, 0.3% protosta-13(17),24-dien-3beta-ol, and 0.7% protosta-16,24-dien-3beta-ol from (3S)-2,3-oxidosqualene
I705C
-
the mutant produces 72% lanosterol, 16% (13alphaH)-isomalabarica-14E,17E,21-dien-3beta-ol, 11.7% (13alphaH)-isomalabarica-14Z,17E,21-dien-3beta-ol, and 0.3% protosta-13(17),24-dien-3beta-ol from (3S)-2,3-oxidosqualene
I705D
-
the mutant produces 37% lanosterol, 32% (13alphaH)-isomalabarica-14E,17E,21-dien-3beta-ol, 30% (13alphaH)-isomalabarica-14Z,17E,21-dien-3beta-ol, and 1% protosta-13(17),24-dien-3beta-ol from (3S)-2,3-oxidosqualene
I705F
-
the mutant produces 25% lanosterol, 21(13alphaH)-isomalabarica-14(26),17E,21-trien-3beta-ol, 6% 17alpha-protosta-20,24-dien-3beta-ol, 42% 17alpha-protosta-20(22),24-dien-3beta-ol from, and 6% protosta-16,24-dien-3beta-ol from (3S)-2,3-oxidosqualene
I705G
-
the mutant produces 35% lanosterol, 23% (13alphaH)-isomalabarica-14E,17E,21-dien-3beta-ol, 34% (13alphaH)-isomalabarica-14Z,17E,21-dien-3beta-ol, 6% protosta-13(17),24-dien-3beta-ol, 1% 17alpha-protosta-20(22),24-dien-3beta-ol and 1% protosta-16,24-dien-3beta-ol from (3S)-2,3-oxidosqualene
I705K
-
the mutant produces 12% lanosterol, 26.5% (13alphaH)-isomalabarica-14E,17E,21-dien-3beta-ol, 53.1% (13alphaH)-isomalabarica-14Z,17E,21-dien-3beta-ol, 7.6% protosta-13(17),24-dien-3beta-ol, and 0.8% protosta-16,24-dien-3beta-ol from (3S)-2,3-oxidosqualene
I705L
-
the mutant produces 100% lanosterol from (3S)-2,3-oxidosqualene (wild-type activity)
I705M
-
the mutant produces 88% lanosterol, 6% (13alphaH)-isomalabarica-14E,17E,21-dien-3beta-ol, and 6% (13alphaH)-isomalabarica-14Z,17E,21-dien-3beta-ol from (3S)-2,3-oxidosqualene
I705N
-
the mutant produces 19% lanosterol, 41% (13alphaH)-isomalabarica-14E,17E,21-dien-3beta-ol, 37% (13alphaH)-isomalabarica-14Z,17E,21-dien-3beta-ol, and 3% protosta-13(17),24-dien-3beta-ol from (3S)-2,3-oxidosqualene
I705P
-
the mutant produces 81.9% lanosterol, 0.1% (13alphaH)-isomalabarica-14E,17E,21-dien-3beta-ol, 15% (13alphaH)-isomalabarica-14Z,17E,21-dien-3beta-ol, and 3% protosta-16,24-dien-3beta-ol from (3S)-2,3-oxidosqualene
I705Q
-
the mutant produces 25% lanosterol, 39% (13alphaH)-isomalabarica-14E,17E,21-dien-3beta-ol, 31% (13alphaH)-isomalabarica-14Z,17E,21-dien-3beta-ol, and 5% protosta-13(17),24-dien-3beta-ol from (3S)-2,3-oxidosqualene
I705S
-
the mutant produces 12% lanosterol, 42% (13alphaH)-isomalabarica-14E,17E,21-dien-3beta-ol, 44% (13alphaH)-isomalabarica-14Z,17E,21-dien-3beta-ol, 1% protosta-13(17),24-dien-3beta-ol, and 1%protosta-16,24-dien-3beta-ol from (3S)-2,3-oxidosqualene
I705T
-
the mutant produces 22% lanosterol, 36% (13alphaH)-isomalabarica-14E,17E,21-dien-3beta-ol, 41% (13alphaH)-isomalabarica-14Z,17E,21-dien-3beta-ol, and 1% protosta-13(17),24-dien-3beta-ol from (3S)-2,3-oxidosqualene
I705V
-
the mutant produces 75% lanosterol, 10% (13alphaH)-isomalabarica-14E,17E,21-dien-3beta-ol, and 15% (13alphaH)-isomalabarica-14Z,17E,21-dien-3beta-ol from (3S)-2,3-oxidosqualene
Q450H
-
site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme produng exclusively achilleol A as product
Q450H/V454I
-
site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme producing lanosterol, (13alphaH)-isomalabarica-14(26),17E,21-trien-3beta-ol, and protosta-16,24-dien-3beta-ol
T384Y
-
site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme producing lanosterol, parkeol, and 9beta-lanosta-7,24-dien-3beta-ol
T384Y/Q450H
-
site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme producing mainly parkeol and very low amounts of 9beta-lanosta-7,24-dien-3beta-ol and lanosterol
T384Y/Q450H/V454I
-
site-directed mutagenesis, the mutant produces exclusively reaction intermediate parkeol but not lanosterol as the sole end product
T384Y/V454I
-
site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme producing mainly parkeol and lower amounts of 9beta-lanosta-7,24-dien-3beta-ol and lanosterol
T509G
-
site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme, overview
V454A
-
results in the production of an additional truncated monocyclic achilleol A
V454G
-
results in the production of an additional truncated monocyclic achilleol A
V454I
-
site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme producing mainly lanosterol and a low amount of (13alphaH)-isomalabarica-14(26),17E,21-trien-3beta-ol
W232A
-
products are 18.1% protosta-12,24-dien-3beta-ol, 47.1% lanosterol, 34.8% parkeol
W232C
-
products are 18% protosta-12,24-dien-3beta-ol, 31.8% lanosterol, 50.2% parkeol
W232D
-
products are 7.9% protosta-12,24-dien-3beta-ol, 84.8% lanosterol, 7.3% parkeol
W232E
-
products are 14% protosta-12,24-dien-3beta-ol, 49.2% lanosterol, 36.8% parkeol
W232F
-
products are 8.4% protosta-12,24-dien-3beta-ol, 74.4% lanosterol, 17,2% parkeol
W232G
-
products are 8.4% protosta-12,24-dien-3beta-ol, 74.4% lanosterol, 17.2% parkeol
W232H
-
products are 27.8% protosta-12,24-dien-3beta-ol, 35% lanosterol, 37.2% parkeol
W232I
-
products are 23.5% protosta-12,24-dien-3beta-ol, 33% lanosterol, 43.5% parkeol
W232K
-
no catalytic activity
W232L
-
products are 14.5% protosta-12,24-dien-3beta-ol, 27.8% lanosterol, 57.7% parkeol
W232M
-
products are 10.9% protosta-12,24-dien-3beta-ol, 40.6% lanosterol, 48.5% parkeol
W232N
-
products are 10.7% protosta-12,24-dien-3beta-ol, 59.5% lanosterol, 29.8% parkeol
W232P
-
products are 4.4% protosta-12,24-dien-3beta-ol, 82.9% lanosterol, 12.7% parkeol
W232Q
-
products are 24.6% protosta-12,24-dien-3beta-ol, 32.1% lanosterol, 43.3% parkeol
W232R
-
no catalytic activity
W232S
-
products are 10.7% protosta-12,24-dien-3beta-ol, 59% lanosterol, 30.3% parkeol
W232T
-
products are 14.3% protosta-12,24-dien-3beta-ol, 61.7% lanosterol, 24% parkeol
W232V
-
products are 19.3% protosta-12,24-dien-3beta-ol, 34% lanosterol, 46.7% parkeol
W232Y
-
products are 4.2% protosta-12,24-dien-3beta-ol, 94.2% lanosterol, 1.6% parkeol
W587F
-
site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme, overview
W587Y
-
site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme, overview
Y510C
-
site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme, overview
Y510F
-
produces lanosterol and isomalabaricatrienol in a 95:5 ratio in strain RXY6, while in strain SMY8, the ratio is 10:90
Y510L
-
site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme, overview
Y510S
-
site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme, overview
Y707A
-
the mutant produces 18.9% (9R,10S)-polypoda-8(26),13E,17E,21-tetraen-3beta-ol, 56.9% lanosterol, 19.9% 9beta-lanosta-7,24-dien-3beta-ol, and 4.3% parkeol from (S)-2,3-oxidosqualene
Y707C
-
the mutant produces 6% (9R,10S)-polypoda-8(26),13E,17E,21-tetraen-3beta-ol, 47.1% lanosterol, 42.6% 9beta-lanosta-7,24-dien-3beta-ol, and 4.3% parkeol from (S)-2,3-oxidosqualene
Y707D
-
the mutant produces 21.8% (9R,10S)-polypoda-8(26),13E,17E,21-tetraen-3beta-ol, 67.3% lanosterol, 7.6% 9beta-lanosta-7,24-dien-3beta-ol, and 3.3% parkeol from (S)-2,3-oxidosqualene
Y707E
-
the mutant produces 12.3% (9R,10S)-polypoda-8(26),13E,17E,21-tetraen-3beta-ol, 70.8% lanosterol, 10.5% 9beta-lanosta-7,24-dien-3beta-ol, and 6.4% parkeol from (S)-2,3-oxidosqualene
Y707F
-
the mutant produces 89.2% lanosterol, 3.1% parkeol, and 7.7% 9beta-lanosta-7,24-dien-3beta-ol from (S)-2,3-oxidosqualene
Y707G
-
the mutant produces 28.3% (9R,10S)-polypoda-8(26),13E,17E,21-tetraen-3beta-ol, 48.2% lanosterol, 16.3% 9beta-lanosta-7,24-dien-3beta-ol, and 7.2% parkeol from (S)-2,3-oxidosqualene
Y707H
-
the mutant produces 83.6% (9R,10S)-polypoda-8(26),13E,17E,21-tetraen-3beta-ol, 4.7% lanosterol, 6.2% 9beta-lanosta-7,24-dien-3beta-ol, and 5.5% parkeol from (S)-2,3-oxidosqualene
Y707I
-
the mutant produces 87.9% lanosterol and 12.1% 9beta-lanosta-7,24-dien-3beta-ol from (S)-2,3-oxidosqualene
Y707K
-
the mutant produces 100% lanosterol from (S)-2,3-oxidosqualene (wild type reaction)
Y707L
-
the mutant produces 85.3% lanosterol and 14.7% 9beta-lanosta-7,24-dien-3beta-ol from (S)-2,3-oxidosqualene
Y707M
-
the mutant produces 100% lanosterol from (S)-2,3-oxidosqualene (wild type reaction)
Y707N
-
the mutant produces 100% lanosterol from (S)-2,3-oxidosqualene (wild type reaction)
Y707P
-
the mutant produces 100% lanosterol from (S)-2,3-oxidosqualene (wild type reaction)
Y707Q
-
the mutant produces 82% (9R,10S)-polypoda-8(26),13E,17E,21-tetraen-3beta-ol, 4.3% lanosterol, and 13.7% parkeol from (S)-2,3-oxidosqualene
Y707S
-
the mutant produces 21.3% (9R,10S)-polypoda-8(26),13E,17E,21-tetraen-3beta-ol, 42.9% lanosterol, 20.5% 9beta-lanosta-7,24-dien-3beta-ol, and 15.3% parkeol from (S)-2,3-oxidosqualene
Y707T
-
the mutant produces 9.8% (9R,10S)-polypoda-8(26),13E,17E,21-tetraen-3beta-ol, 65.1% lanosterol, 13.5% 9beta-lanosta-7,24-dien-3beta-ol, and 11.6% parkeol from (S)-2,3-oxidosqualene
Y707V
-
the mutant produces 100% lanosterol from (S)-2,3-oxidosqualene (wild type reaction)
Y707W
-
the mutant produces 100% lanosterol from (S)-2,3-oxidosqualene (wild type reaction)
Y707X
-
the mutant produces (9R,10S)-polypoda-8(26),13E,17E,21-tetraen-3beta-ol, in addition to lanosterol, parkeol, and 9beta-lanosta-7,24-dien-3beta-ol from (S)-2,3-oxidosqualene
C457D
-
less active than the wild-type
-
C457D/E526A
-
decrease in enzyme functionality
-
C457D/E526C
-
very sensitive to the thiol-reacting agent dodecylmaleimide, specific activity and thermal stability are severely reduced
-
C457D/E526D
-
no effect on catalytic avtivity
-
C457D/E526Q
-
no effect on catalytic avtivity
-
F699I
-
the mutant produces lanosterol (100%) as the wild type enzyme
-
F699L
-
the mutant produces lanosterol (100%) as the wild type enzyme
-
F699N
-
nonviable mutant, does not produce lanosterol, but protosta-13(17)-dien-3beta-ol (55%), malabarica-14E,17E,21-trien-3beta-ol (5%), 17alpha-protosta-20,24-dien-3beta-ol (24%), and (17Z)-protosta-17(20),24-dien-3beta-ol (16%)
-
F699T
-
the mutant produces lanosterol (less than 0.2%) and protosta-13(17)-dien-3beta-ol (above 99.8%)
-
F699X
-
TKW14[pERG7F699X] site-saturated mutants allow for ergosterol-independent growth, with the exception of Leu, Ile, His, Met, Pro, and Thr substitutions
-
F699C
-
inactive
F699C
-
the mutant is inactive
F699M
-
the mutant produces lanosterol (13%), (17Z)-protosta-17(20),24-dien-3beta-ol (46%), protosta-13(17)-dien-3beta-ol (70%), malabarica-14E,17E,21-trien-3beta-ol (7%), 17alpha-protosta-20,24-dien-3beta-ol (1%), (17Z)-protosta-17(20),24-dien-3beta-ol (10%), (13alphaH)-isomalabarica-14E,17E,21-dien-3 beta-ol (17%), and (13alphaH)-isomalabarica-14Z,17E,21-dien-3beta-ol (18%)
F699M
-
the mutant produces 1% lanosterol, 17% (13alphaH)-isomalabarica-14E,17E,21-dien-3beta-ol, 18% (13alphaH)-isomalabarica-14Z,17E,21-dien-3beta-ol, 46% protosta-13(17),24-dien-3beta-ol, 1% 17alpha-protosta-20,24-dien-3beta-ol, 10% protosta-17(20),24-dien-3beta-ol and 7% malabarica-14E,17E,21-trien-3beta-ol from (3S)-2,3-oxidosqualene
F699T
-
the mutant produces novel protosta-13(17),24-dien-3beta-ol as the sole truncated rearrangement product from (S)-2,3-oxidosqualene
F699T
-
the mutant produces protosta-13(17),24-dien-3beta-ol from (S)-2,3-oxidosqualene
F699T
-
the mutant produces lanosterol (less than 0.2%) and protosta-13(17)-dien-3beta-ol (above 99.8%)
F699T
-
the mutant produces less than 0.2% lanosterol and more than 99.8% protosta-13(17),24-dien-3beta-ol from (3S)-2,3-oxidosqualene
H234W/Y510W
-
mutant produces achilleol A from (S)-2,3-oxidosqualene
H234W/Y510W
-
site-directed mutagenesis, the double mutation alters the ERG7 function to achilleol A synthase activity and generates achilleol A as the sole product
Y510A
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produces lanosterol, parkeol and achilleol A in a 39:34:27 ratio
Y510A
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site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme, overview
Y510H
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incomplete cyclization, produces achilleol (45%), lanosterol (42%), parkeol (9%), and isomalabaricatrienol (4%)
Y510H
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site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme, overview
Y510K
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inactive
Y510K
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fails to maintain cell viability in the absence of ergosterol, in the presence of ergosterol the mutant produces achilleol A and camelliol C in a ratio of 86:14
Y510K
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site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme, overview
Y510W
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inactive
Y510W
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fails to maintain cell viability in the absence of ergosterol, in the presence of ergosterol the mutant produces achilleol A and camelliol C in a ratio of 96:4
Y510W
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site-directed mutagenesis, the mutant shows an altered product profile compared to the wild-type enzyme, overview
additional information
no visible morphological phenotypes are observed in enzyme disruption mutant and no differences in sterol profiles between the wild type and mutant are detected. Enzyme gene complements enzyme-deficient yeast strain
additional information
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an oxidosqualene:protostadienol cyclase mutant in which the C-terminal residues 702APPGGMR708 are replaced with 702NKSCAIS708 efficiently produces a 1:1 mixture of lanosterol and parkeol
additional information
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only polar side-chain group substitutions of F445 genetically complement yeast viability and produce spatially related product diversity
additional information
overexpression of Arabidopsis thaliana AtLAS in leaves via transformation by Agrobacterium tumefaciens strain EHA 105, where the endogenic StLAS-like is not expressed, results in increased steroidal glycoalkaloid level and reduced phytosterol level in the leaves, while the steroidal glycoalkaloid level in the tuber flesh is reduced
additional information
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overexpression of Arabidopsis thaliana AtLAS in leaves via transformation by Agrobacterium tumefaciens strain EHA 105, where the endogenic StLAS-like is not expressed, results in increased steroidal glycoalkaloid level and reduced phytosterol level in the leaves, while the steroidal glycoalkaloid level in the tuber flesh is reduced