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 (Comment) | Organism |
---|---|
gene asuE1, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Streptomyces nodosus |
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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces nodosus | D7P5V0 | subsp. asukaensis | - |
Streptomyces nodosus ATCC 29757 | D7P5V0 | subsp. asukaensis | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chomatography | Streptomyces nodosus |
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 | Comment | Organism |
---|---|---|
asuE1 | - |
Streptomyces nodosus |
flavin-dependent protoasukamycin hydroxylase | - |
Streptomyces nodosus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Streptomyces nodosus |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Streptomyces nodosus |
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 | 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 |