Literature summary for 4.1.99.5 extracted from

Jia, C.; Li, M.; Li, J.; Zhang, J.; Zhang, H.; Cao, P.; Pan, X.; Lu, X.; Chang, W.
Structural insights into the catalytic mechanism of aldehyde-deformylating oxygenases (2015), Protein Cell, 6, 55-67 .

Cloned(Commentary)

Cloned (Comment)	Organism
gene Synpcc7942_1593, sequence comparisons, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)	Synechococcus elongatus PCC 7942 = FACHB-805

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme in both its iron-free and iron-bound forms or with bound substrate, and of mutants Y122F and F86Y/F87Y, sitting drop vapor diffusion method, from reservoir solution of 0.2 mol/l Mg2+, 0.1 mol/l Tris, pH 8.5, 30% w/v PEG 4000, for the iron-bound crystals, 4 mM ferrous ammonium sulfate is added to SeADO protein solutions right before crystallization, and for the substrate-bound enzyme, 0.2mol/l L-proline, 0.1mol/l HEPES, pH 7.1, 25% w/v PEG 1500 is used, 18°C, X-ray diffraction structure determination and analysis at 1.71-2.9 A resolution	Synechococcus elongatus PCC 7942 = FACHB-805

Crystallization (Comment)

Organism

purified recombinant enzyme in both its iron-free and iron-bound forms or with bound substrate, and of mutants Y122F and F86Y/F87Y, sitting drop vapor diffusion method, from reservoir solution of 0.2 mol/l Mg2+, 0.1 mol/l Tris, pH 8.5, 30% w/v PEG 4000, for the iron-bound crystals, 4 mM ferrous ammonium sulfate is added to SeADO protein solutions right before crystallization, and for the substrate-bound enzyme, 0.2mol/l L-proline, 0.1mol/l HEPES, pH 7.1, 25% w/v PEG 1500 is used, 18°C, X-ray diffraction structure determination and analysis at 1.71-2.9 A resolution

Synechococcus elongatus PCC 7942 = FACHB-805

Protein Variants

Protein Variants	Comment	Organism
F86Y/F87Y	site-directed mutagenesis, structure comparison with the wild-type enzyme	Synechococcus elongatus PCC 7942 = FACHB-805
additional information	enzyme structure analysis, comparisons of wild-type and mutant structures, overview	Synechococcus elongatus PCC 7942 = FACHB-805
Y122F	site-directed mutagenesis, structure comparison with the wild-type enzyme	Synechococcus elongatus PCC 7942 = FACHB-805

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Fe2+	di-iron center, Glu144 is one of the iron-coordinating residues and plays a vital role in the catalytic reaction of cADO. The helix, in which Glu144 resides, exhibits two distinct conformations that correlate with the different binding states of the di-iron center in cADO structures. The highly labile feature of cADO di-iron center seems to be responsible for the low enzymatic activity. Six conservative amino acids, loctaed in two EX28-29EX2H motifs, from four helices (Glu32 from helix H1, Glu115 from helix H4, Glu60 and His63 from helix H2, and Glu144 and His147 from helix H5) act as metal ligands. The WT0 structure is characterized by losing the di-iron cluster and by exhibiting a distorted conformation of helix H5. This structure is likely to represent the inactive state of SeADO, as it has lost its cofactor iron	Synechococcus elongatus PCC 7942 = FACHB-805

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
a long-chain aldehyde + O2 + 2 NADPH + 2 H+	Synechococcus elongatus PCC 7942 = FACHB-805	-	an alkane + formate + H2O + 2 NADP+	-	?
a long-chain aldehyde + O2 + 2 NADPH + 2 H+	Synechococcus elongatus PCC 7942 = FACHB-805 R2	-	an alkane + formate + H2O + 2 NADP+	-	?

Organism

Organism	UniProt	Comment	Textmining
Synechococcus elongatus PCC 7942 = FACHB-805	Q54764	i.e. Synechococcus elongates strain PCC 7942	-
Synechococcus elongatus PCC 7942 = FACHB-805 R2	Q54764	i.e. Synechococcus elongates strain PCC 7942	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration, followed by ultrafiltration	Synechococcus elongatus PCC 7942 = FACHB-805

Reaction

Reaction	Comment	Organism	Reaction ID
octadecanal + O2 + 2 NADPH + 2 H+ = heptadecane + formate + H2O + 2 NADP+	enzyme structures representing the different states during catalytic reaction	Synechococcus elongatus PCC 7942 = FACHB-805	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
a long-chain aldehyde + O2 + 2 NADPH + 2 H+	-	Synechococcus elongatus PCC 7942 = FACHB-805	an alkane + formate + H2O + 2 NADP+	-	?
a long-chain aldehyde + O2 + 2 NADPH + 2 H+	-	Synechococcus elongatus PCC 7942 = FACHB-805 R2	an alkane + formate + H2O + 2 NADP+	-	?
additional information	the in vitro reaction catalyzed by cADO requires both the dioxygen as co-substrate and the presence of a reducing system, which provides four electrons per turnover and can either be biological (ferredoxin, ferredoxin reductase, and NADPH) or chemical (phenazine methosulfate and NADH)	Synechococcus elongatus PCC 7942 = FACHB-805	?	-	?
additional information	the in vitro reaction catalyzed by cADO requires both the dioxygen as co-substrate and the presence of a reducing system, which provides four electrons per turnover and can either be biological (ferredoxin, ferredoxin reductase, and NADPH) or chemical (phenazine methosulfate and NADH)	Synechococcus elongatus PCC 7942 = FACHB-805 R2	?	-	?
n-heptanal + O2 + 2 NADPH + 2 H+	-	Synechococcus elongatus PCC 7942 = FACHB-805	n-hexane + formate + H2O + 2 NADP+	-	?
n-heptanal + O2 + 2 NADPH + 2 H+	-	Synechococcus elongatus PCC 7942 = FACHB-805 R2	n-hexane + formate + H2O + 2 NADP+	-	?

Subunits

Subunits	Comment	Organism
More	enzyme structure analysis, comparisons of wild-type and mutant structures, overview	Synechococcus elongatus PCC 7942 = FACHB-805

Synonyms

Synonyms	Comment	Organism
aldehyde-deformylating oxygenase	-	Synechococcus elongatus PCC 7942 = FACHB-805
cADO	-	Synechococcus elongatus PCC 7942 = FACHB-805
erial aldehyde-deformylating oxygenase	-	Synechococcus elongatus PCC 7942 = FACHB-805
SeADO	-	Synechococcus elongatus PCC 7942 = FACHB-805
Synpcc7942_1593	-	Synechococcus elongatus PCC 7942 = FACHB-805

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Synechococcus elongatus PCC 7942 = FACHB-805

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.2	-	assay at	Synechococcus elongatus PCC 7942 = FACHB-805

Cofactor

Cofactor	Comment	Organism	Structure
NADPH	-	Synechococcus elongatus PCC 7942 = FACHB-805

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to the superfamily of ferritin-like di-iron proteins with conserved sequence of two EX28-29EX2H motifs	Synechococcus elongatus PCC 7942 = FACHB-805
metabolism	cyanobacterial aldehyde-deformylating oxygenase (cADO), which catalyzes the conversion of Cn fatty aldehyde to its corresponding Cn-1 alk(a/e)ne, is a key enzyme in fatty alk(a/e)ne biosynthesis pathway	Synechococcus elongatus PCC 7942 = FACHB-805
additional information	Glu144, one of the iron-coordinating residues, plays a vital role in the catalytic reaction of cADO. The helix, in which Glu144 resides, exhibits two distinct conformations that correlate with the different binding states of the di-iron center in cADO structures. Enzyme structure analysis, comparisons of wild-type and mutant structures, overview. A continuous tube-shaped non-protein electron density, resembling a lipid molecule, is observed close to the di-iron center in all structures but the Y122F structure, a hydrophobic substrate channel in SeADO is described	Synechococcus elongatus PCC 7942 = FACHB-805
physiological function	cyanobacterial aldehyde-deformylating oxygenase (cADO) catalyzes the conversion of Cn fatty aldehyde to its corresponding Cn-1 alk(a/e)ne with low activity due to a highly labile feature of cADO di-iron center	Synechococcus elongatus PCC 7942 = FACHB-805