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 (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 | 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 | 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 | 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 | 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 (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 | 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 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | 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+ | - |
? | |
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 | Comment | Organism |
---|---|---|
More | enzyme structure analysis, comparisons of wild-type and mutant structures, overview | Synechococcus elongatus PCC 7942 = FACHB-805 |
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 Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Synechococcus elongatus PCC 7942 = FACHB-805 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.2 | - |
assay at | Synechococcus elongatus PCC 7942 = FACHB-805 |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | - |
Synechococcus elongatus PCC 7942 = FACHB-805 |
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 |