Literature summary for 1.14.14.1 extracted from

Jungmann, V.; Molnar, I.; Hammer, P.E.; Hill, D.S.; Zirkle, R.; Buckel, T.G.; Buckel, D.; Ligon, J.M.; Pachlatko, J.P.
Biocatalytic conversion of avermectin to 4"-oxo-avermectin: characterization of biocatalytically active bacterial strains and of cytochrome p450 monooxygenase enzymes and their genes (2005), Appl. Environ. Microbiol., 71, 6968-6976.

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Cloned(Commentary)

Cloned (Comment)	Organism
gene family Ema, DNA and amino acid sequence determination and analysis	Streptomyces sp.
gene family ema, DNA and amino acid sequence determination and analysis, overexpression of His-tagged ema1 and ema16 in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli and Streptomyces lividans	Streptomyces tubercidicus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	kinetic analysis of recombinant enzymes	Streptomyces tubercidicus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
avermectin + [reduced NADPH-hemoprotein reductase] + O2	Streptomyces sp.	4''-oxo-avermectin is a key intermediate in the manufacture of the agriculturally important insecticide emamectin benzoate from the natural product avermectin, overview	4''-oxo-avermectin + [oxidized NADPH-hemoprotein reductase] + H2O	-	?
avermectin + [reduced NADPH-hemoprotein reductase] + O2	Streptomyces tubercidicus	4''-oxo-avermectin is a key intermediate in the manufacture of the agriculturally important insecticide emamectin benzoate from the natural product avermectin, overview	4''-oxo-avermectin + [oxidized NADPH-hemoprotein reductase] + H2O	-	?

Organism

Organism	UniProt	Comment	Textmining
Streptomyces sp.	-	several strains, Ema genes	-
Streptomyces tubercidicus	-	strains R-922 and I-1529, ema genes, and genes cyp229 and cyp230	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged Ema1 and Ema16 from Escherichia coli to near-homogeneity by a two-step purification procedure	Streptomyces tubercidicus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.001	0.06	purified recombinant Ema1 and Ema16 proteins	Streptomyces tubercidicus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
avermectin + [reduced NADPH-hemoprotein reductase] + O2	4''-oxo-avermectin is a key intermediate in the manufacture of the agriculturally important insecticide emamectin benzoate from the natural product avermectin, overview	Streptomyces sp.	4''-oxo-avermectin + [oxidized NADPH-hemoprotein reductase] + H2O	-	?
avermectin + [reduced NADPH-hemoprotein reductase] + O2	4''-oxo-avermectin is a key intermediate in the manufacture of the agriculturally important insecticide emamectin benzoate from the natural product avermectin, overview	Streptomyces tubercidicus	4''-oxo-avermectin + [oxidized NADPH-hemoprotein reductase] + H2O	-	?
avermectin + [reduced NADPH-hemoprotein reductase] + O2	avermectin oxidation activity	Streptomyces sp.	4''-oxo-avermectin + [oxidized NADPH-hemoprotein reductase] + H2O	-	?
avermectin + [reduced NADPH-hemoprotein reductase] + O2	avermectin oxidation activity	Streptomyces tubercidicus	4''-oxo-avermectin + [oxidized NADPH-hemoprotein reductase] + H2O	-	?

Synonyms

Synonyms	Comment	Organism
Cyp	-	Streptomyces sp.
Cyp	-	Streptomyces tubercidicus
cytochrome P450 monooxygenase	-	Streptomyces sp.
cytochrome P450 monooxygenase	-	Streptomyces tubercidicus
Ema	-	Streptomyces tubercidicus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assya at	Streptomyces tubercidicus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.048	-	avermectin	pH 7.0, 30°C, recombinant Ema1	Streptomyces tubercidicus

pH Optimum

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

Cofactor

Cofactor	Comment	Organism	Structure
Ferredoxin	-	Streptomyces sp.
Ferredoxin	-	Streptomyces tubercidicus

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Release 2023.1 (January 2023)