Literature summary for 4.1.1.98 extracted from

Do, H.; Lee, C.W.; Han, S.J.; Lee, S.G.; Kim, H.J.; Park, H.; Lee, J.H.
Purification, crystallization and preliminary X-ray crystallographic studies of FMN-bound and FMN-free forms of aromatic acid decarboxylase (CpsUbiX) from the psychrophilic bacterium Colwellia psychrerythraea 34H (2014), Acta Crystallogr. Sect. F, 70, 215-220 .

Cloned(Commentary)

Cloned (Comment)	Organism
gene ubiX, recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)	Colwellia psychrerythraea

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His6-tagged wild-type (FMN-bound) and V47S mutant (FMN-free) enzymes, hanging drop vapour diffusion method, mixing of 800 nl of 21 mg/ml protein solution with 800 nl of reservoir solution containing 200 mM sodium chloride, 100 mM potassium phosphate monobasic/sodium phosphate dibasic, pH 5.8, and 11% w/v PEG 8000, for the wild-type enzyme and 0.1 M trisodium citrate, pH 5.4, 0.5 M ammonium sulfate, and 1.2 M lithium sulfate for the mutant enzyme, 20°C, 3 days, method optimization, X-ray diffraction structrue determination and analysis at 2.0 A and 1.76 A resolution, respectively	Colwellia psychrerythraea

Crystallization (Comment)

Organism

purified recombinant His6-tagged wild-type (FMN-bound) and V47S mutant (FMN-free) enzymes, hanging drop vapour diffusion method, mixing of 800 nl of 21 mg/ml protein solution with 800 nl of reservoir solution containing 200 mM sodium chloride, 100 mM potassium phosphate monobasic/sodium phosphate dibasic, pH 5.8, and 11% w/v PEG 8000, for the wild-type enzyme and 0.1 M trisodium citrate, pH 5.4, 0.5 M ammonium sulfate, and 1.2 M lithium sulfate for the mutant enzyme, 20°C, 3 days, method optimization, X-ray diffraction structrue determination and analysis at 2.0 A and 1.76 A resolution, respectively

Colwellia psychrerythraea

Protein Variants

Protein Variants	Comment	Organism
V47S	site-directed mutagenesis, the single mutation results in a loss of FMN binding, resulting in the production of FMN-free CpsUbiX protein. The mutation is likely to destabilize FMN-protein interactions without affecting the overall structural folding. The mutant enzyme has an FMN:protein molar ratio of 0.086:1. Wild-type CpsUbiX protein has a yellowish colour, while the V47S mutant does not	Colwellia psychrerythraea

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
256300	-	recombinant wild-type enzyme	Colwellia psychrerythraea

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
a 4-hydroxy-3-polyprenylbenzoate	Colwellia psychrerythraea	-	a 2-polyprenylphenol + CO2	-	?

Organism

Organism	UniProt	Comment	Textmining
Colwellia psychrerythraea	Q489U8	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, gel filtration, and ultrafiltration	Colwellia psychrerythraea

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
a 4-hydroxy-3-polyprenylbenzoate	-	Colwellia psychrerythraea	a 2-polyprenylphenol + CO2	-	?
additional information	analysis of the conformational changes upon FMN binding and the enzymatic mechanism at the molecular level, overview	Colwellia psychrerythraea	?	-	?

Subunits

Subunits	Comment	Organism
dodecamer	12 * 22552, sequence calculation	Colwellia psychrerythraea

Synonyms

Synonyms	Comment	Organism
CpsUbiX	-	Colwellia psychrerythraea
flavin mononucleotide-dependent aromatic acid decarboxylase	-	Colwellia psychrerythraea
FMN-dependent aromatic acid decarboxylase	-	Colwellia psychrerythraea
ubiX	-	Colwellia psychrerythraea

Cofactor

Cofactor	Comment	Organism	Structure
FMN	wild-type CpsUbiX has an FMN:protein molar ratio of 1.028:1. The protein with a V47S single mutation cannot bind FMN and has an FMN:protein molar ratio of 0.086:1	Colwellia psychrerythraea

pI Value

Organism	Comment	pI Value Maximum	pI Value
Colwellia psychrerythraea	sequence calculation	-	5.78

General Information

General Information	Comment	Organism
evolution	UbiX belongs to the FMN-dependent decarboxylase family	Colwellia psychrerythraea
metabolism	a set of enzymes, UbiA, UbiB, UbiC, UbiD, UbiE, UbiF, UbiG, UbiH and UbiX, catalyze the synthesis of ubiquinone from the compound chorismate. Enzyme UbiX catalyzes and regulates ubiquinone biosynthesis	Colwellia psychrerythraea