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Literature summary for 5.4.3.11 extracted from

  • Ratnayake, N.; Liu, N.; Kuhn, L.; Walker, K.
    Ring-substituted beta-arylalanines for probing substituent effects on the isomerization reaction catalyzed by an aminomutase (2014), ACS Catal., 4, 3077-3090 .
No PubMed abstract available
Search for more literature for 5.4.3.11

Application

Application Comment Organism
synthesis utility of the methylidene imidazolone-dependent Pantoea agglomerans phenylalanine aminomutase (PaPAM) for making non-natural beta-amino acids, substrate specificity of PaPAM with several commercially available (S)-arylalanine substrates, overview Pantoea agglomerans

Cloned(Commentary)

Cloned (Comment) Organism
gene papam, recombinant expression of C-terminally His6-tagged PaPAM in Escherichia coli strain BL21(DE3) Pantoea agglomerans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten kinetics Pantoea agglomerans
0.027
-
 3-fluoro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.029
-
 4-fluoro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.073
-
 2-fluoro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.088
-
 2-methyl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.132
-
 3-thiophen-2-yl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.163
-
 4-methyl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.164
-
 2-methoxy-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.168
-
 L-phenylalanine pH 8.0, 31°C Pantoea agglomerans
0.204
-
 3-methyl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.337
-
 3-thiophen-3-yl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.339
-
 3-bromo-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.415
-
 3-furan-2-yl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.43
-
 3-nitro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.432
-
 3-chloro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.491
-
 4-chloro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.525
-
 4-bromo-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.752
-
 4-nitro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.99
-
 3-methoxy-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
1.187
-
 4-methoxy-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-phenylalanine Pantoea agglomerans
-
 D-beta-phenylalanine
-
 ?

Organism

Organism UniProt Comment Textmining
Pantoea agglomerans Q84FL5 i.e. Enterobacter agglomerans or Erwinia herbicola
-

Purification (Commentary)

Purification (Comment) Organism
recombinant C-terminally His6-tagged PaPAM from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, and ultrafiltration, to over 95% purity Pantoea agglomerans

Reaction

Reaction Comment Organism Reaction ID
L-phenylalanine = D-beta-phenylalanine reaction mechanism, detailed overview. Proposed elimination mechanisms for the displacement of the NH2-MIO adduct. The final reaction sequence of the MIO-dependent aminomutases involves an alpha,beta-addition reaction, where the NH2-MIO and a proton add across the double bond of the acrylate intermediate. Alternatively, PaPAM can use a stepwise addition sequence where the nucleophile (NH2-MIO) couples to form a 1,4-Michael adduct. This conjugate addition route benefits from an electropositive (delta+) Cbeta by delocalizing the Pi-electrons toward the carboxylate of the substrate Pantoea agglomerans
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-fluoro-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(2-fluorophenyl)propanoic acid
-
 ?
2-methoxy-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(2-methoxyphenyl)propanoic acid
-
 ?
2-methyl-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(o-tolyl)propanoic acid
-
 ?
3-bromo-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(3-bromophenyl)propanoic acid
-
 ?
3-chloro-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(3-chlorophenyl)propanoic acid
-
 ?
3-fluoro-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(3-fluorophenyl)propanoic acid
-
 ?
3-furan-2-yl-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(furan-2-yl)propanoic acid
-
 ?
3-methoxy-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(3-methoxyphenyl)propanoic acid
-
 ?
3-methyl-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(3-methylphenyl)propanoic acid
-
 ?
3-nitro-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(3-nitrophenyl)propanoic acid
-
 ?
3-thiophen-2-yl-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(thiophen-2-yl)propanoic acid
-
 ?
3-thiophen-3-yl-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(thiophen-3-yl)propanoic acid
-
 ?
4-bromo-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(4-bromophenyl)propanoic acid
-
 ?
4-chloro-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(4-chlorophenyl)propanoic acid
-
 ?
4-fluoro-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(4-fluorophenyl)propanoic acid
-
 ?
4-methoxy-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(4-methoxyphenyl)propanoic acid
-
 ?
4-methyl-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(4-methylphenyl)propanoic acid
-
 ?
4-nitro-L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-(4-nitrophenyl)propanoic acid
-
 ?
L-phenylalanine
-
 Pantoea agglomerans D-beta-phenylalanine
-
 ?
L-phenylalanine
-
 Pantoea agglomerans (S)-3-amino-3-phenylpropanoic acid
-
 ?
additional information enzyme substrate specificity, detailed overview. Interactions the most significant are between the substrates and residues Phe428, Val108, Leu421, Leu104, Gln456, and Tyr. Electronic effects of ortho-, para-, and meta-substituents. The enzyme produces a different amount of cinnamate as side-product dependent on the substrate, e.g. the product pool catalyzed by PaPAM from 13 contains more cinnamate analogue (21.7%) compared to that made from other m-substituted substrates 3-bromo-L-phenylalanine, 3-chloro-L-phenylalanine, 3-nitro-L-phenylalanine, and 3-methoxy-L-phenylalanine that contain between 1.0% and 7.4%. Relationship between PaPAM-substrate interaction energies, flexibility, and Km. Compounds 2-bromo-L-phenylalanine, 2-chloro-L-phenylalanine, and 2-nitro-L-phenylalanine are no substrates Pantoea agglomerans ?
-
 ?

Subunits

Subunits Comment Organism
? x * 59000, about, recombinant His-tagged enzyme, SDS-PAGE Pantoea agglomerans

Synonyms

Synonyms Comment Organism
admH
-
 Pantoea agglomerans
methylidene imidazolone-dependent phenylalanine aminomutase
-
 Pantoea agglomerans
MIO-dependent phenylalanine ammonia lyase
-
 Pantoea agglomerans
PaPAM
-
 Pantoea agglomerans
phenylalanine aminomutase
-
 Pantoea agglomerans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
31
-
 assay at Pantoea agglomerans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.003
-
 2-methoxy-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.013
-
 4-methyl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.022
-
 4-methoxy-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.022
-
 2-fluoro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.023
-
 4-fluoro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.026
-
 3-thiophen-2-yl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.031
-
 3-fluoro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.038
-
 4-nitro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.045
-
 4-bromo-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.053
-
 4-chloro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.058
-
 3-methyl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.064
-
 2-methyl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.143
-
 3-thiophen-3-yl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.146
-
 3-nitro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.209
-
 3-methoxy-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.236
-
 3-furan-2-yl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.301
-
 L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.42
-
 3-bromo-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.484
-
 3-chloro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Pantoea agglomerans

Cofactor

Cofactor Comment Organism Structure
4-methylidene-1H-imidazol-5(4H)-one MIO, prosthetic group, dependent on, proposed elimination mechanisms for the displacement of the NH2-MIO adduct. Formation of an NH2-MIO adduct, where the amino group of the substrate is covalently attached to the enzyme during alpha/beta-isomerization Pantoea agglomerans

General Information

General Information Comment Organism
evolution the enzyme belongs to a class I lyase-like superfamily of catalysts, along with other MIO-dependent aminomutases Pantoea agglomerans
additional information enzyme structure-activity analysis and modeling of substrate-PaPAM structural interactions, overview Pantoea agglomerans
physiological function PaPAM makes the (3S)-beta-phenylalanine antipode on the biosynthetic pathway to the antibiotic andrimid starting from (S)-beta-phenylalanine via (S)-alpha-phenylalanine Pantoea agglomerans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.019
-
 4-methoxy-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.02
-
 2-methoxy-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.05
-
 4-nitro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.051
-
 L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.082
-
 4-methyl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.09
-
 4-bromo-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.11
-
 4-chloro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.19
-
 3-methyl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.19
-
 3-thiophen-2-yl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.21
-
 3-methoxy-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.31
-
 2-fluoro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.34
-
 3-nitro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.428
-
 3-thiophen-3-yl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.588
-
 3-furan-2-yl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.73
-
 2-methyl-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
0.79
-
 4-fluoro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
1.12
-
 3-chloro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
1.2
-
 3-fluoro-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans
1.24
-
 3-bromo-L-phenylalanine pH 8.0, 31°C, recombinant enzyme Pantoea agglomerans