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Literature summary for 4.3.1.24 extracted from

  • Hsieh, L.S.; Yeh, C.S.; Pan, H.C.; Cheng, C.Y.; Yang, C.C.; Lee, P.D.
    Cloning and expression of a phenylalanine ammonia-lyase gene (BoPAL2) from Bambusa oldhamii in Escherichia coli and Pichia pastoris (2010), Protein Expr. Purif., 71, 224-230.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Pichia pastoris strain X-33 and Escherichia coli Top10 cells Bambusa oldhamii
expression in Escherichia coli an Pichia pastoris Bambusa oldhamii
expression in Escherichia coli and Pichia pastoris Bambusa oldhamii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.25
-
L-phenylalanine recombinant enzyme expressed in Escherichia coli Bambusa oldhamii
0.25
-
L-phenylalanine recombinant enzyme expressed in Escherichia coli, at 37°C and pH 8.5 Bambusa oldhamii
0.25
-
L-Phe recombinant protein from Escherichia coli, pH 8.5, 37°C Bambusa oldhamii
0.331
-
L-phenylalanine recombinant enzyme expressed in Pichi pastoris Bambusa oldhamii
0.331
-
L-phenylalanine recombinant enzyme expressed in Pichia pastoris, at 37°C and pH 8.5 Bambusa oldhamii
0.331
-
L-Phe recombinant protein from Pichia pastoris, pH 8.5, 37°C Bambusa oldhamii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
76970
-
subunit, calculated from amino acid sequence Bambusa oldhamii
76970
-
4 * 76970, calculated, 4 * 80000, SDS-PAGE of recombinant protein with His-tag Bambusa oldhamii
80000
-
4 * 80000, SDS-PAGE Bambusa oldhamii
80000
-
4 * 76970, calculated, 4 * 80000, SDS-PAGE of recombinant protein with His-tag Bambusa oldhamii
330000
-
gel filtration Bambusa oldhamii
330000
-
SDS-PAGE Bambusa oldhamii
330000
-
PAGE and gel filtration Bambusa oldhamii

Organism

Organism UniProt Comment Textmining
Bambusa oldhamii
-
-
-
Bambusa oldhamii C0LL35
-
-
Bambusa oldhamii C0LL35 isoform Pal2
-

Purification (Commentary)

Purification (Comment) Organism
HiTrap affinity chromatography Bambusa oldhamii
Ni-IDA column column chromatography Bambusa oldhamii
recombinant protein from Escherichia coli Bambusa oldhamii

Source Tissue

Source Tissue Comment Organism Textmining
shoot
-
Bambusa oldhamii
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.19
-
crude recombinant extract from Escherichia coli, at 37°C and pH 8.5 Bambusa oldhamii
0.41
-
crude recombinant extract from Pichia pastoris, at 37°C and pH 8.5 Bambusa oldhamii
3.76
-
recombinant enzyme from Pichia pastoris, after 9.1fold purification, at 37°C and pH 8.5 Bambusa oldhamii
3.76
-
recombinant protein from Pichia pastoris, pH 8.5, 37°C Bambusa oldhamii
3.77
-
recombinant enzyme expressed in Escherichia coli Bambusa oldhamii
3.77
-
recombinant protein from Escherichia coli, pH 8.5, 37°C Bambusa oldhamii
3.78
-
recombinant enzyme from Escherichia coli, after 19.4fold purification, at 37°C and pH 8.5 Bambusa oldhamii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-Phe
-
Bambusa oldhamii (E)-cinnamate + NH3
-
?
L-phenylalanine
-
Bambusa oldhamii (E)-cinnamate + NH3
-
?
L-phenylalanine assay at pH 8.5, 37°C, reaction terminated by addition of HCl Bambusa oldhamii (E)-cinnamate + NH3
-
?
additional information L-Tyr is a poor substrate Bambusa oldhamii ?
-
?

Subunits

Subunits Comment Organism
homotetramer 4 * 80000, SDS-PAGE Bambusa oldhamii
tetramer 4 * 80000, SDS-PAGE Bambusa oldhamii
tetramer 4 * 76970, calculated, 4 * 80000, SDS-PAGE of recombinant protein with His-tag Bambusa oldhamii

Synonyms

Synonyms Comment Organism
EC 4.3.1.5 formerly Bambusa oldhamii
PAL
-
Bambusa oldhamii
PAL2
-
Bambusa oldhamii
phenylalanine ammonia-lyase
-
Bambusa oldhamii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
recombinant enzyme expressed in Escherichia coli Bambusa oldhamii
50 60
-
Bambusa oldhamii
60
-
recombinant enzyme expressed in Pichi pastoris Bambusa oldhamii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
10.12
-
L-phenylalanine recombinant enzyme expressed in Escherichia coli Bambusa oldhamii
10.12
-
L-phenylalanine recombinant enzyme expressed in Escherichia coli, at 37°C and pH 8.5 Bambusa oldhamii
10.12
-
L-Phe recombinant protein from Escherichia coli, pH 8.5, 37°C Bambusa oldhamii
16.04
-
L-phenylalanine recombinant enzyme expressed in Pichi pastoris Bambusa oldhamii
16.04
-
L-phenylalanine recombinant enzyme expressed in Pichia pastoris, at 37°C and pH 8.5 Bambusa oldhamii
16.04
-
L-Phe recombinant protein from Pichia pastoris, pH 8.5, 37°C Bambusa oldhamii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
recombinant enzyme expressed in Escherichia coli Bambusa oldhamii
8.5 9
-
Bambusa oldhamii
9
-
recombinant enzyme expressed in Pichi pastoris Bambusa oldhamii

Cofactor

Cofactor Comment Organism Structure
additional information PAL2 sequence containes a conserved active site motif, the Ala-Ser-Gly triad, which can be converted into a 3,5-dihydro-5-methylidine-4H-imidazol-4-one prosthetic group Bambusa oldhamii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.05
-
L-phenylalanine recombinant enzyme expressed in Escherichia coli Bambusa oldhamii
4.85
-
L-phenylalanine recombinant enzyme expressed in Pichi pastoris Bambusa oldhamii
40.5
-
L-Phe recombinant protein from Escherichia coli, pH 8.5, 37°C Bambusa oldhamii
48.5
-
L-Phe recombinant protein from Pichia pastoris, pH 8.5, 37°C Bambusa oldhamii