Literature summary for 1.13.11.88 extracted from

Ryu, J.Y.; Seo, J.; Park, S.; Ahn, J.H.; Chong, Y.; Sadowsky, M.J.; Hur, H.G.
Characterization of an isoeugenol monooxygenase (Iem) from Pseudomonas nitroreducens Jin1 that transforms isoeugenol to vanillin (2013), Biosci. Biotechnol. Biochem., 77, 289-294 .

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Activating Compound

Activating Compound	Comment	Organism	Structure
additional information	not inhibitory or activating at 1 mM: Mg(II), Mn(II), Mo(II), NADH, FAD, or ascorbic acid, EDTA, 1,10-phenanthroline, and Tiron	Pseudomonas nitroreducens

Cloned(Commentary)

Cloned (Comment)	Organism
expression in ERscherichia coli	Pseudomonas nitroreducens

Protein Variants

Protein Variants	Comment	Organism
H167A	mutation of conserved His residue, involved in binding of Fe2+. Loss of catalytic activity	Pseudomonas nitroreducens
H205A	mutation of conserved His residue, 6.8% residual catalytic activity	Pseudomonas nitroreducens
H218A	mutation of conserved His residue, involved in binding of Fe2+. Loss of catalytic activity	Pseudomonas nitroreducens
H282A	mutation of conserved His residue, involved in binding of Fe2+. Loss of catalytic activity	Pseudomonas nitroreducens

Inhibitors

Inhibitors	Comment	Organism	Structure
Co2+	1 mM, 40% of initial activity	Pseudomonas nitroreducens
additional information	not inhibitory or activating at 1 mM: Mg(II), Mn(II), Mo(II), NADH, FAD, or ascorbic acid, EDTA, 1,10-phenanthroline, and Tiron	Pseudomonas nitroreducens
Ni2+	1 mM, 49% of initial activity	Pseudomonas nitroreducens
Zn2+	1 mM, 35% of initial activity	Pseudomonas nitroreducens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.12	-	isoeugenol	pH 8.0, 30°C	Pseudomonas nitroreducens

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Fe2+	presence of 0.7 mol iron per mol of enzyme	Pseudomonas nitroreducens
Fe3+	1 mM, 114% of initial activity	Pseudomonas nitroreducens

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas nitroreducens	C3VA26	-	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
4.2	-	pH 8.0, 30°C	Pseudomonas nitroreducens

Storage Stability

Storage Stability	Organism
-70°C, 20mM Tris/HCl, 10% glycerol, and 1mM dithiothreitol, pH 8.0, stable for several months	Pseudomonas nitroreducens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
isoeugenol + O2	-	Pseudomonas nitroreducens	vanillin + acetaldehyde	-	?
additional information	oxidative cleavage of isoeugenol by Iem is catalyzed via a monooxygenation reaction, and incorporation of the oxygen atom from O2 into vanillin is preferred over incorporation from water. Iem exhibits no activity toward any other of the phenylpropanoid and styrene compounds tested	Pseudomonas nitroreducens	?	-	?

Subunits

Subunits	Comment	Organism
?	x * 54000, SDS-PAGE	Pseudomonas nitroreducens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	-	Pseudomonas nitroreducens

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
20	-	stable for 30 min	Pseudomonas nitroreducens
30	-	30 min, 7% loss of activity	Pseudomonas nitroreducens
50	-	30 min, complete loss of activity	Pseudomonas nitroreducens

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
3.8	-	isoeugenol	pH 8.0, 30°C	Pseudomonas nitroreducens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
9	-	-	Pseudomonas nitroreducens

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Release 2023.1 (January 2023)