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Literature summary for 1.14.13.215 extracted from

  • Rui, Z.; Sandy, M.; Jung, B.; Zhang, W.
    Tandem enzymatic oxygenations in biosynthesis of epoxyquinone pharmacophore of manumycin-type metabolites (2013), Chem. Biol., 20, 879-887.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
protein AsuE2 despite being a single-component monooxygenase, AsuE1 activity is enhanced by AsuE2, a pathway-specific flavin reductase, UniProt-ID D7P5W0, AsuE2 is encoded in the asu gene cluster. Enzyme AsuE2 might assist the catalysis of AsuE1 by providing a reduced form of flavin in Streptomyces nodosus Streptomyces nodosus

Cloned(Commentary)

Cloned (Comment) Organism
gene asuE1, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Streptomyces nodosus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0123
-
NADH in presence of 0.01 mM FAD, recombinant enzyme, pH 7.0, 30°C Streptomyces nodosus
0.0213
-
protoasukamycin in presence of 0.01 mM FAD, recombinant enzyme, pH 7.0, 30°C Streptomyces nodosus
1.84
-
NADPH in presence of 0.01 mM FAD, recombinant enzyme, pH 7.0, 30°C Streptomyces nodosus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
protoasukamycin + NADH + H+ + O2 Streptomyces nodosus enzyme AsuE1 is a flavin-dependent protoasukamycin hydroxylase, that belongs to the family of single-component monooxygenases: a single enzyme catalyzes both the reduction of flavin by NADPH, and the hydroxylation using molecular oxygen and reduced flavin cofactor 4-hydroxyprotoasukamycin + NAD+ + H2O
-
?
protoasukamycin + NADH + H+ + O2 Streptomyces nodosus ATCC 29757 enzyme AsuE1 is a flavin-dependent protoasukamycin hydroxylase, that belongs to the family of single-component monooxygenases: a single enzyme catalyzes both the reduction of flavin by NADPH, and the hydroxylation using molecular oxygen and reduced flavin cofactor 4-hydroxyprotoasukamycin + NAD+ + H2O
-
?

Organism

Organism UniProt Comment Textmining
Streptomyces nodosus D7P5V0 subsp. asukaensis
-
Streptomyces nodosus ATCC 29757 D7P5V0 subsp. asukaensis
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chomatography Streptomyces nodosus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
protoasukamycin + NADH + H+ + O2 enzyme AsuE1 is a flavin-dependent protoasukamycin hydroxylase, that belongs to the family of single-component monooxygenases: a single enzyme catalyzes both the reduction of flavin by NADPH, and the hydroxylation using molecular oxygen and reduced flavin cofactor Streptomyces nodosus 4-hydroxyprotoasukamycin + NAD+ + H2O
-
?
protoasukamycin + NADH + H+ + O2 NADH is the preferred cofactor, it reduces the flavin that gets oxidized in the hydroxylation of protoasukamycin, reaction chain overview Streptomyces nodosus 4-hydroxyprotoasukamycin + NAD+ + H2O
-
?
protoasukamycin + NADH + H+ + O2 enzyme AsuE1 is a flavin-dependent protoasukamycin hydroxylase, that belongs to the family of single-component monooxygenases: a single enzyme catalyzes both the reduction of flavin by NADPH, and the hydroxylation using molecular oxygen and reduced flavin cofactor Streptomyces nodosus ATCC 29757 4-hydroxyprotoasukamycin + NAD+ + H2O
-
?
protoasukamycin + NADH + H+ + O2 NADH is the preferred cofactor, it reduces the flavin that gets oxidized in the hydroxylation of protoasukamycin, reaction chain overview Streptomyces nodosus ATCC 29757 4-hydroxyprotoasukamycin + NAD+ + H2O
-
?
protoasukamycin + NADPH + H+ + O2
-
Streptomyces nodosus 4-hydroxyprotoasukamycin + NADP+ + H2O
-
?
protoasukamycin + NADPH + H+ + O2
-
Streptomyces nodosus ATCC 29757 4-hydroxyprotoasukamycin + NADP+ + H2O
-
?

Synonyms

Synonyms Comment Organism
asuE1
-
Streptomyces nodosus
flavin-dependent protoasukamycin hydroxylase
-
Streptomyces nodosus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Streptomyces nodosus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.113
-
NADH in presence of 0.01 mM FAD, recombinant enzyme, pH 7.0, 30°C Streptomyces nodosus
0.324
-
NADPH in presence of 0.01 mM FAD, recombinant enzyme, pH 7.0, 30°C Streptomyces nodosus
16.67
-
protoasukamycin in presence of 0.01 mM FAD, recombinant enzyme, pH 7.0, 30°C Streptomyces nodosus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Streptomyces nodosus

Cofactor

Cofactor Comment Organism Structure
FAD
-
Streptomyces nodosus
FMN enzyme AsuE2 contains a trace amount of FMN at a molar ratio of 1:2000 (FMN:AsuE2) upon purification from Escherichia coli Streptomyces nodosus
additional information enzyme AsuE1 is a flavin-dependent protoasukamycin hydroxylase, that displays no significant preference toward flavin cofactors, because FAD, FMN, and riboflavin are utilized efficiently in the catalysis Streptomyces nodosus
NADH preferential utilization of NADH by AsuE2 Streptomyces nodosus
NADPH
-
Streptomyces nodosus
riboflavin
-
Streptomyces nodosus

General Information

General Information Comment Organism
evolution enzyme AsuE1 belongs to the family of single-component monooxygenases: a single enzyme catalyzes both the reduction of flavin by NADPH, and the hydroxylation using molecular oxygen and reduced flavin cofactor Streptomyces nodosus
malfunction disruption of asuE2 significantly decreases the yield of asukamycin and results in the accumulation of protoasukamycin Streptomyces nodosus
metabolism AsuE1 and AsuE3, both single-component flavin-dependent monooxygenases, catalyze the exquisite regio- and enantiospecific postpolyketide synthase (PKS) assembly oxygenations. AsuE1 installs a hydroxyl group on the 3,4-AHB ring to form a 4-hydroxyquinone moiety, which is epoxidized by AsuE3 to yield the epoxyquinone functionality. Despite being a single-component monooxygenase, AsuE1 activity is elicited by AsuE2, a pathway-specific flavin reductase Streptomyces nodosus