Literature summary for 1.13.11.75 extracted from

Sui, X.; Kiser, P.D.; Che, T.; Carey, P.R.; Golczak, M.; Shi, W.; von Lintig, J.; Palczewski, K.
Analysis of carotenoid isomerase activity in a prototypical carotenoid cleavage enzyme, apocarotenoid oxygenase (ACO) (2014), J. Biol. Chem., 289, 12286-12299 .

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Cloned(Commentary)

Cloned (Comment)	Organism
gene sll1541, recombinant expression in Escherichia coli strain BL21	Synechocystis sp. PCC 6803

Crystallization (Commentary)

Crystallization (Comment)	Organism
native enzyme in the absence of apocarotenoid substrate, hanging drop vapor diffusion method, mixing of 0.001 ml of 10 mg/ml protein in 25 mM HEPES-NaOH, pH 7.0, 1 mM dithiothreitol, and 0.8% w/v hexaethylene glycol monooctyl ether or 0.02% w/v Triton X-100, with 0.001 ml of reservoir solution containing 0.1 M BTP-HCl, pH 6.0, 2223% w/v PEG 3350, 0.2 M NH4Cl, and 1 mM MnCl2, 8°C, 3-4 days, X-ray diffraction structure determination and analysis. ACO crystallization strongly inhibits the apocarotenoid oxygenase activity of the enzyme	Synechocystis sp. PCC 6803

Crystallization (Comment)

Organism

native enzyme in the absence of apocarotenoid substrate, hanging drop vapor diffusion method, mixing of 0.001 ml of 10 mg/ml protein in 25 mM HEPES-NaOH, pH 7.0, 1 mM dithiothreitol, and 0.8% w/v hexaethylene glycol monooctyl ether or 0.02% w/v Triton X-100, with 0.001 ml of reservoir solution containing 0.1 M BTP-HCl, pH 6.0, 2223% w/v PEG 3350, 0.2 M NH4Cl, and 1 mM MnCl2, 8°C, 3-4 days, X-ray diffraction structure determination and analysis. ACO crystallization strongly inhibits the apocarotenoid oxygenase activity of the enzyme

Synechocystis sp. PCC 6803

Inhibitors

Inhibitors	Comment	Organism
CHAPS	slight inhibition	Synechocystis sp. PCC 6803
CYMAL-4	slight inhibition	Synechocystis sp. PCC 6803
hexaethylene glycol monooctyl ether	strongly inhibits ACO activity even at concentrations below its critical micelle concentration	Synechocystis sp. PCC 6803
additional information	the enzyme ACO is inhibited by linear polyoxyethylene detergents, mechanism, overview. ACO crystallization strongly inhibits the apocarotenoid oxygenase activity of the enzyme. PEG 3350 has little influence on enzyme activity up to a concentration of10% w/v	Synechocystis sp. PCC 6803
tetraethylene glycol monooctyl ether	strongly inhibits ACO activity even at concentrations below its critical micelle concentration	Synechocystis sp. PCC 6803

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
additional information	-	additional information	steady-state kinetics, Michaelis-Menten kinetics	Synechocystis sp. PCC 6803
0.121	-	all-trans-8'-apo-beta-carotenal	recombinant enzyme, pH 7.0, 28°C	Synechocystis sp. PCC 6803
0.127	-	all-trans-8'-apo-beta-carotenal	recombinant enzyme, pH 7.0, 28°C, in presence of 0.1 CMC C8E6	Synechocystis sp. PCC 6803

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
all-trans-8'-apocarotenol + O2	Synechocystis sp. PCC 6803	-	all-trans-retinal + (2E,4E,6E)-8-hydroxy-2,6-dimethylocta-2,4,6-trienal	-	?

Organism

Organism	UniProt	Comment	Textmining
Synechocystis sp. PCC 6803	P74334	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant enzyme from Escherichia coli strain BL21 by ammonium sulfate fractionation and gel filtration	Synechocystis sp. PCC 6803

Reaction

Reaction	Comment	Organism	Reaction ID
all-trans-8'-apo-beta-carotenal + O2 = all-trans-retinal + (2E,4E,6E)-2,6-dimethylocta-2,4,6-trienedial	the enzyme utilizes a non-heme iron center to oxidatively cleave a carbon-carbon double bond of a carotenoid substrate	Synechocystis sp. PCC 6803	a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.47	-	purified recombinant enzyme, pH 7.0, 28°C	Synechocystis sp. PCC 6803

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
all-trans-8'-apo-beta-carotenol + O2	-	Synechocystis sp. PCC 6803	all-trans-retinal + (2E,4E,6E)-8-hydroxy-2,6-dimethylocta-2,4,6-trienal	-	?
all-trans-8'-apocarotenol + O2	-	Synechocystis sp. PCC 6803	all-trans-retinal + (2E,4E,6E)-8-hydroxy-2,6-dimethylocta-2,4,6-trienal	-	?
additional information	enzyme ACO exclusively produces all-trans-retinal from all-trans-apo-8'-carotenol. ACO cleaves but does not isomerize all-trans-8'-apocarotenol. Raman spectroscopic study, overview. During the entire reaction, spectroscopic signals for 13-cis-retinal or the 13,14'-di-cis-8-apocarotenol intermediate are not detected	Synechocystis sp. PCC 6803	?	-	?

Synonyms

Synonyms	Comment	Organism
ACO	-	Synechocystis sp. PCC 6803
apocarotenoid oxygenase	-	Synechocystis sp. PCC 6803
sll1541	-	Synechocystis sp. PCC 6803

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
28	-	assay at	Synechocystis sp. PCC 6803

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	assay at	Synechocystis sp. PCC 6803

General Information

General Information	Comment	Organism
evolution	ACO is a classical non-isomerizing member of the carotenoid cleavage enzyme (CCE) family	Synechocystis sp. PCC 6803
additional information	active site structure, overview	Synechocystis sp. PCC 6803