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Literature summary for 5.1.1.5 extracted from

  • Kuan, Y.; Kao, C.; Chen, C.; Chen, C.; Hu, H.; Hsu, W.
    Biochemical characterization of a novel lysine racemase from Proteus mirabilis BCRC10725 (2011), Process Biochem., 46, 1914-1920.
No PubMed abstract available

Cloned(Commentary)

Cloned (Comment) Organism
gene lyr, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Proteus mirabilis

Protein Variants

Protein Variants Comment Organism
S394C site-directed mutagenesis, the mutant shows 1.5fold increased activity with L-arginine compared to the wild-type enzyme Proteus mirabilis
S394N site-directed mutagenesis, the mutant shows about 2.2fold increased activity with L-arginine compared to the wild-type enzyme Proteus mirabilis
S394T site-directed mutagenesis, the mutant shows 1.8fold increased activity with L-arginine compared to the wild-type enzyme Proteus mirabilis
S394Y site-directed mutagenesis, the mutant shows 2.1fold increased activity with L-arginine compared to the wild-type enzyme Proteus mirabilis

Inhibitors

Inhibitors Comment Organism Structure
hydroxylamine completely inhibition of the enzyme's L-Lys racemization at 5 mM, partially restored by the addition of pyridoxal 5'-phosphate Proteus mirabilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1 7 L-arginine pH 8.0, 50┬░C, recombinant mutant S394T Proteus mirabilis
1 2.5 L-arginine pH 8.0, 50┬░C, recombinant mutant S394C Proteus mirabilis
2 6 L-lysine pH 8.0, 50┬░C, recombinant mutant S394C Proteus mirabilis
2 6 L-lysine pH 8.0, 50┬░C, recombinant mutant S394N Proteus mirabilis
14.9
-
L-arginine pH 8.0, 50┬░C, recombinant wild-type enzyme Proteus mirabilis
17.1
-
L-arginine pH 8.0, 50┬░C, recombinant mutant S394N Proteus mirabilis
17.8
-
L-arginine pH 8.0, 50┬░C, recombinant mutant S394Y Proteus mirabilis
21.8
-
L-lysine pH 8.0, 50┬░C, recombinant wild-type enzyme Proteus mirabilis
25.2
-
L-lysine pH 8.0, 50┬░C, recombinant mutant S394T Proteus mirabilis
26.5
-
L-lysine pH 8.0, 50┬░C, recombinant mutant S394Y Proteus mirabilis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-lysine Proteus mirabilis
-
D-lysine
-
r
L-lysine Proteus mirabilis BCRC10725
-
D-lysine
-
r

Organism

Organism UniProt Comment Textmining
Proteus mirabilis M4GGR9 gene lyr
-
Proteus mirabilis BCRC10725 M4GGR9 gene lyr
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) Proteus mirabilis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [┬Ámol/min/mg] Specific Activity Maximum [┬Ámol/min/mg] Comment Organism
568
-
purified recombinant His-tagged enzyme, substrate L-arginine, pH 8.0, 50┬░C Proteus mirabilis
887
-
purified recombinant His-tagged mutant S394Y, substrate L-lysine, pH 8.0, 50┬░C Proteus mirabilis
980
-
purified recombinant His-tagged mutant S394C, substrate L-arginine, pH 8.0, 50┬░C Proteus mirabilis
1043
-
purified recombinant His-tagged mutant S394T, substrate L-arginine, pH 8.0, 50┬░C Proteus mirabilis
1185
-
purified recombinant His-tagged mutant S394Y, substrate L-arginine, pH 8.0, 50┬░C Proteus mirabilis
1200
-
purified recombinant His-tagged mutant S394N, substrate L-arginine, pH 8.0, 50┬░C Proteus mirabilis
1633
-
purified recombinant His-tagged mutant S394N, substrate L-lysine, pH 8.0, 50┬░C Proteus mirabilis
2356
-
purified recombinant His-tagged mutant S394C, substrate L-lysine, pH 8.0, 50┬░C Proteus mirabilis
2828
-
purified recombinant His-tagged enzyme, substrate L-lysine, pH 8.0, 50┬░C Proteus mirabilis
3127
-
purified recombinant His-tagged mutant S394T, substrate L-lysine, pH 8.0, 50┬░C Proteus mirabilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-arginine
-
Proteus mirabilis D-arginine
-
r
L-arginine
-
Proteus mirabilis BCRC10725 D-arginine
-
r
L-lysine
-
Proteus mirabilis D-lysine
-
r
L-lysine
-
Proteus mirabilis BCRC10725 D-lysine
-
r
additional information enzyme residue S394 residue plays a crucial role in determining the preference of Lyr to lysine and arginine. No activity by wild-type and mutant enzyme with alanine, methionine, histidine, tryptophan, leucine, glutamine, asparagine and serine Proteus mirabilis ?
-
?
additional information enzyme residue S394 residue plays a crucial role in determining the preference of Lyr to lysine and arginine. No activity by wild-type and mutant enzyme with alanine, methionine, histidine, tryptophan, leucine, glutamine, asparagine and serine Proteus mirabilis BCRC10725 ?
-
?

Synonyms

Synonyms Comment Organism
lyr
-
Proteus mirabilis

Temperature Optimum [┬░C]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
50
-
recombinant His-tagged enzyme, substrate L-lysine Proteus mirabilis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
10.83
-
L-arginine pH 8.0, 50┬░C, recombinant wild-type enzyme Proteus mirabilis
14.93
-
L-lysine pH 8.0, 50┬░C, recombinant mutant S394Y Proteus mirabilis
16.2
-
L-arginine pH 8.0, 50┬░C, recombinant mutant S394C Proteus mirabilis
17.45
-
L-arginine pH 8.0, 50┬░C, recombinant mutant S394T Proteus mirabilis
18.75
-
L-arginine pH 8.0, 50┬░C, recombinant mutant S394N Proteus mirabilis
19.67
-
L-arginine pH 8.0, 50┬░C, recombinant mutant S394Y Proteus mirabilis
31.25
-
L-lysine pH 8.0, 50┬░C, recombinant mutant S394N Proteus mirabilis
50.68
-
L-lysine pH 8.0, 50┬░C, recombinant mutant S394C Proteus mirabilis
55.43
-
L-lysine pH 8.0, 50┬░C, recombinant wild-type enzyme Proteus mirabilis
62.5
-
L-lysine pH 8.0, 50┬░C, recombinant mutant S394T Proteus mirabilis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8 9 recombinant His-tagged enzyme, substrate L-lysine Proteus mirabilis

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate dependent on Proteus mirabilis