BRENDA - Enzyme Database

Alteration of product specificity of Rhodobacter sphaeroides phytoene desaturase by directed evolution

Wang, C.W.; Liao, J.C.; J. Biol. Chem. 276, 41161-41164 (2001)

Data extracted from this reference:

Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.3.5.6
E508G
random mutagenesis, the mutation has little effect on the enzyme activity
Rhodobacter sphaeroides
1.3.5.6
F166I
random mutagenesis, a beneficial mutation that helps the conversion of product from neurosporene to lycopene
Rhodobacter sphaeroides
1.3.5.6
F220S
random mutagenesis, the mutation has little effect on the enzyme activity
Rhodobacter sphaeroides
1.3.5.6
H12Q
random mutagenesis, the mutation has little effect on the enzyme activity
Rhodobacter sphaeroides
1.3.5.6
L148H
random mutagenesis, the beneficial mutation is highly increasing the production of lycopene to over 80%
Rhodobacter sphaeroides
1.3.5.6
M402T
random mutagenesis, the mutation reduces the lycopene production rate
Rhodobacter sphaeroides
1.3.5.6
additional information
alteration of product specificity of Rhodobacter sphaeroides phytoene desaturase by directed evolution combining random mutagenesis and site-directed mutagenesis, DNA sequence determinations. Beneficial mutations increase the percent of lycopene production to 90%; introduction of stop mutations, M1, M2 and M3
Rhodobacter sphaeroides
1.3.5.6
V68D
random mutagenesis, a beneficial mutation that helps the conversion of product from neurosporene to lycopene
Rhodobacter sphaeroides
1.3.99.28
F166I
mutation changes the product of phytoene desaturation from neurosporene to lycopene
Rhodobacter sphaeroides
1.3.99.28
H12Q
mutation has little effect on the product formation
Rhodobacter sphaeroides
1.3.99.28
M402T
mutation has a negative effect on percent lycopene production
Rhodobacter sphaeroides
1.3.99.28
additional information
directed evolution is used to change the product of Rhodobacter sphaeroides phytoene desaturase (crtI gene product), a neurosporene-producing enzyme, to lycopene. Two generations of random mutagenesis are performed, from which three positive mutants are isolated and sequenced. Site-directed mutagenesis is used to determine the effect of each amino acid change
Rhodobacter sphaeroides
1.3.99.28
V68D
mutation changes the product of phytoene desaturation from neurosporene to lycopene
Rhodobacter sphaeroides
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.3.5.6
phytoene + quinone
Rhodobacter sphaeroides
via phytofluene and zeta-carotene
7,9,9'-tricis-neurosporene + quinol
-
-
?
1.3.99.28
15-cis-phytoene + 3 acceptor
Rhodobacter sphaeroides
the enzyme is involved in carotenoid biosynthesis
all-trans-neurosporene + 3 reduced acceptor
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.3.5.6
Rhodobacter sphaeroides
-
gene crtI
-
1.3.99.28
Rhodobacter sphaeroides
-
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.3.5.6
7,9,9'-tricis-neurosporene + a quinone
mutant activity
712351
Rhodobacter sphaeroides
7,9,7',9'-tetracis-lycopene + a quinol
-
-
-
?
1.3.5.6
phytoene + quinone
via phytofluene and zeta-carotene
712351
Rhodobacter sphaeroides
7,9,9'-tricis-neurosporene + quinol
-
-
-
?
1.3.99.28
15-cis-phytoene + 3 acceptor
the enzyme is involved in carotenoid biosynthesis
712351
Rhodobacter sphaeroides
all-trans-neurosporene + 3 reduced acceptor
-
-
-
?
1.3.99.28
phytoene + acceptor
-
712351
Rhodobacter sphaeroides
neurosporene + reduced acceptor
-
-
-
?
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.3.5.6
E508G
random mutagenesis, the mutation has little effect on the enzyme activity
Rhodobacter sphaeroides
1.3.5.6
F166I
random mutagenesis, a beneficial mutation that helps the conversion of product from neurosporene to lycopene
Rhodobacter sphaeroides
1.3.5.6
F220S
random mutagenesis, the mutation has little effect on the enzyme activity
Rhodobacter sphaeroides
1.3.5.6
H12Q
random mutagenesis, the mutation has little effect on the enzyme activity
Rhodobacter sphaeroides
1.3.5.6
L148H
random mutagenesis, the beneficial mutation is highly increasing the production of lycopene to over 80%
Rhodobacter sphaeroides
1.3.5.6
M402T
random mutagenesis, the mutation reduces the lycopene production rate
Rhodobacter sphaeroides
1.3.5.6
additional information
alteration of product specificity of Rhodobacter sphaeroides phytoene desaturase by directed evolution combining random mutagenesis and site-directed mutagenesis, DNA sequence determinations. Beneficial mutations increase the percent of lycopene production to 90%; introduction of stop mutations, M1, M2 and M3
Rhodobacter sphaeroides
1.3.5.6
V68D
random mutagenesis, a beneficial mutation that helps the conversion of product from neurosporene to lycopene
Rhodobacter sphaeroides
1.3.99.28
F166I
mutation changes the product of phytoene desaturation from neurosporene to lycopene
Rhodobacter sphaeroides
1.3.99.28
H12Q
mutation has little effect on the product formation
Rhodobacter sphaeroides
1.3.99.28
M402T
mutation has a negative effect on percent lycopene production
Rhodobacter sphaeroides
1.3.99.28
additional information
directed evolution is used to change the product of Rhodobacter sphaeroides phytoene desaturase (crtI gene product), a neurosporene-producing enzyme, to lycopene. Two generations of random mutagenesis are performed, from which three positive mutants are isolated and sequenced. Site-directed mutagenesis is used to determine the effect of each amino acid change
Rhodobacter sphaeroides
1.3.99.28
V68D
mutation changes the product of phytoene desaturation from neurosporene to lycopene
Rhodobacter sphaeroides
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.3.5.6
phytoene + quinone
Rhodobacter sphaeroides
via phytofluene and zeta-carotene
7,9,9'-tricis-neurosporene + quinol
-
-
?
1.3.99.28
15-cis-phytoene + 3 acceptor
Rhodobacter sphaeroides
the enzyme is involved in carotenoid biosynthesis
all-trans-neurosporene + 3 reduced acceptor
-
-
?
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.3.5.6
7,9,9'-tricis-neurosporene + a quinone
mutant activity
712351
Rhodobacter sphaeroides
7,9,7',9'-tetracis-lycopene + a quinol
-
-
-
?
1.3.5.6
phytoene + quinone
via phytofluene and zeta-carotene
712351
Rhodobacter sphaeroides
7,9,9'-tricis-neurosporene + quinol
-
-
-
?
1.3.99.28
15-cis-phytoene + 3 acceptor
the enzyme is involved in carotenoid biosynthesis
712351
Rhodobacter sphaeroides
all-trans-neurosporene + 3 reduced acceptor
-
-
-
?
1.3.99.28
phytoene + acceptor
-
712351
Rhodobacter sphaeroides
neurosporene + reduced acceptor
-
-
-
?
General Information
EC Number
General Information
Commentary
Organism
1.3.5.6
evolution
approximately 10% of known phytoene desaturases, as in Rhodobacter, produce neurosporene, whereas the rest produce lycopene. These two types of enzymes, although similar in function, have relatively low similarity, below 60%, in terms of nucleotide or amino acid sequence. Phytoene desaturase encoded by gene crtI is a neurosporene-producing enzyme
Rhodobacter sphaeroides
1.3.5.6
physiological function
phytoene desaturase encoded by crtI catalyzes the desaturation of phytoene in the carotenoid biosynthesis pathway, approximately 10% of known phytoene desaturases, as in Rhodobacter, produce neurosporene, whereas the rest produce lycopene, mechanism controlling the product specificity, phytoene desaturase encoded by gene crtI is a neurosporene-producing enzyme, overview
Rhodobacter sphaeroides
1.3.99.28
physiological function
involved in carotenoid biosynthesis
Rhodobacter sphaeroides
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.3.5.6
evolution
approximately 10% of known phytoene desaturases, as in Rhodobacter, produce neurosporene, whereas the rest produce lycopene. These two types of enzymes, although similar in function, have relatively low similarity, below 60%, in terms of nucleotide or amino acid sequence. Phytoene desaturase encoded by gene crtI is a neurosporene-producing enzyme
Rhodobacter sphaeroides
1.3.5.6
physiological function
phytoene desaturase encoded by crtI catalyzes the desaturation of phytoene in the carotenoid biosynthesis pathway, approximately 10% of known phytoene desaturases, as in Rhodobacter, produce neurosporene, whereas the rest produce lycopene, mechanism controlling the product specificity, phytoene desaturase encoded by gene crtI is a neurosporene-producing enzyme, overview
Rhodobacter sphaeroides
1.3.99.28
physiological function
involved in carotenoid biosynthesis
Rhodobacter sphaeroides