Literature summary for 4.3.1.24 extracted from

Wang, L.; Gamez, A.; Archer, H.; Abola, E.E.; Sarkissian, C.N.; Fitzpatrick, P.; Wendt, D.; Zhang, Y.; Vellard, M.; Bliesath, J.; Bell, S.M.; Lemontt, J.F.; Scriver, C.R.; Stevens, R.C.
Structural and biochemical characterization of the therapeutic Anabaena variabilis phenylalanine ammonia lyase (2008), J. Mol. Biol., 380, 623-635.

Application

Application	Comment	Organism
pharmacology	the ability of PAL to catalyze the conversion of L-Phe into nontoxic compounds in the absence of additional cofactors leads to its use as a therapeutic agent for the treatment of phenylketonuria	Trichormus variabilis

Crystallization (Commentary)

Crystallization (Comment)	Organism
C503S/C565S mutant PAL, sitting drop vapor diffusion method, 25°C, mixing of equal volumes of 300 nl of protein and reservoir solutions, the latter containing 12-20% PEG 1500 and 100 mM succinic acid, sodium dihydrogen phosphate, and glycine, pH 7.0, versus 0.07 ml of reservoir solution, cryoprotection by 25% glucose, X-ray diffraction structure determination and analysis at 1.87 A resolution, modeling	Trichormus variabilis

Crystallization (Comment)

Organism

C503S/C565S mutant PAL, sitting drop vapor diffusion method, 25°C, mixing of equal volumes of 300 nl of protein and reservoir solutions, the latter containing 12-20% PEG 1500 and 100 mM succinic acid, sodium dihydrogen phosphate, and glycine, pH 7.0, versus 0.07 ml of reservoir solution, cryoprotection by 25% glucose, X-ray diffraction structure determination and analysis at 1.87 A resolution, modeling

Trichormus variabilis

Protein Variants

Protein Variants	Comment	Organism
C503S	site-directed mutagenesis	Trichormus variabilis
C503S/C565S	site-directed mutagenesis, structure analysis and comparison to the wild-type enzyme, the mutant shows increased activity and resistance to proteases compared to the wild-type enzyme	Trichormus variabilis
C565S	site-directed mutagenesis	Trichormus variabilis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.05	-	phenylalanine	C503S/C565S mutant PAL	Trichormus variabilis
0.06	-	L-phenylalanine	wild-type PAL	Trichormus variabilis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
L-phenylalanine	Trichormus variabilis	-	(E)-cinnamate + NH3	-	?

Organism

Organism	UniProt	Comment	Textmining
Trichormus variabilis	-	-	-

Reaction

Reaction	Comment	Organism	Reaction ID
L-phenylalanine = trans-cinnamate + NH3	analysis of active-site structure analysis and reaction mechanism in which the amino group of the phenylalanine substrate is attacked directly by the 4-methylidene-imidazole-5-one prosthetic group, the reaction proceeds through a carbanion intermediate, Tyr78 acts as a general base, helix-to-loop conformational switch in the helices flanking the inner active-site loop that regulates accessibility of the active site, modeling, overview	Trichormus variabilis	a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
1.7	-	wild-type PAL	Trichormus variabilis
2.2	-	C503S/C565S mutant PAL	Trichormus variabilis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
L-phenylalanine	-	Trichormus variabilis	(E)-cinnamate + NH3	-	?
additional information	structure-function relationship, with helix-loop conformational switch, overview	Trichormus variabilis	?	-	?

Synonyms

Synonyms	Comment	Organism
PAL	-	Trichormus variabilis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Trichormus variabilis

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
4	-	phenylalanine	C503S/C565S mutant PAL	Trichormus variabilis
4.6	-	L-phenylalanine	wild-type PAL	Trichormus variabilis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	8.5	C503S/C565S mutant PAL	Trichormus variabilis

pH Range

pH Minimum	pH Maximum	Comment	Organism
4	12	pH-dependent activity profile	Trichormus variabilis

Cofactor

Cofactor	Comment	Organism	Structure
4-methylidene-imidazole-5-one	deamination of L-Phe is dependent upon a 4-methylidene-imidazole-5-one prosthetic group that is produced by the posttranslational condensation of a sequential Ala-Ser-Gly triad in the enzyme	Trichormus variabilis