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Literature summary for 1.5.1.3 extracted from

  • Takahashi, H.; Arai, M.; Takenawa, T.; Sota, H.; Xie, Q.H.; Iwakura, M.
    Stabilization of hyperactive dihydrofolate reductase by cyanocysteine-mediated backbone cyclization (2007), J. Biol. Chem., 282, 9420-9429.
    View publication on PubMed
Search for more literature for 1.5.1.3

Protein Variants

Protein Variants Comment Organism
M1A/M16N/M20L/M42Y/C85A/M92F/C152S mutant ANLYF carries seven amino acid substitutions that result in a methionine- and cysteine-free mutant enzyme with decreased stability. Mutant ANLYF show very high activity and may be stabilized by backbone cyclization via a cyanocysteine-mediated intramolecular ligation reaction, without loss of its high activity Escherichia coli
additional information construction of mutants based on mutant M1A/M16N/M20L/M42Y/C85A/M92F/C152S, i.e. ANLYF. Linear ANLYF-Gn is ANLYF with the N- and C-terminal extensions of Ala-(Gly)n-3 and -(Gly)2-Cys-Ala residues, respectively, with n=6-8, and circular ANLYF-Gn is ANLYF with the C and N termini cross-linked by the linker of the -Gly-Gly-Ala-(Gly)n-3 sequence, n=6-8 Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0012
-
 7,8-dihydrofolate wild-type, 15°C, pH 7.0 Escherichia coli
0.0062
-
 7,8-dihydrofolate mutant linear ANLYF-G6, 15°C, pH 7.0 Escherichia coli
0.0083
-
 7,8-dihydrofolate mutant M1A/M16N/M20L/M42Y/C85A/M92F/C152S, 15°C, pH 7.0 Escherichia coli
0.0086
-
 7,8-dihydrofolate mutant linear ANLYF-G8, 15°C, pH 7.0 Escherichia coli
0.0115
-
 7,8-dihydrofolate mutant circular ANLYF-G6, 15°C, pH 7.0 Escherichia coli
0.013
-
 7,8-dihydrofolate mutant circular ANLYF-G7, 15°C, pH 7.0 Escherichia coli
0.014
-
 7,8-dihydrofolate mutant circular ANLYF-G8, 15°C, pH 7.0 Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli P0ABQ4
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
7,8-dihydrofolate + NADPH + H+
-
 Escherichia coli 5,6,7,8-tetrahydrofolate + NADP+
-
 ?

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40.9
-
 melting temperature, mutant linear ANLYF-G7 Escherichia coli
42
-
 melting temperature, mutant M1A/M16N/M20L/M42Y/C85A/M92F/C152S Escherichia coli
44.3
-
 melting temperature, mutant linear ANLYF-G8 Escherichia coli
44.5
-
 melting temperature, mutant linear ANLYF-G6 Escherichia coli
49.3
-
 melting temperature, mutant circular ANLYF-G8 Escherichia coli
49.5
-
 melting temperature, mutant circular ANLYF-G6 Escherichia coli
50.2
-
 melting temperature, wild-type Escherichia coli
51.3
-
 melting temperature, mutant circular ANLYF-G7 Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.3
-
 7,8-dihydrofolate wild-type, 15°C, pH 7.0 Escherichia coli
34.8
-
 7,8-dihydrofolate mutant circular ANLYF-G8, 15°C, pH 7.0 Escherichia coli
36.7
-
 7,8-dihydrofolate mutant linear ANLYF-G6, 15°C, pH 7.0 Escherichia coli
38.7
-
 7,8-dihydrofolate mutant circular ANLYF-G7, 15°C, pH 7.0 Escherichia coli
40.5
-
 7,8-dihydrofolate mutant M1A/M16N/M20L/M42Y/C85A/M92F/C152S, 15°C, pH 7.0 Escherichia coli
40.8
-
 7,8-dihydrofolate mutant linear ANLYF-G8, 15°C, pH 7.0 Escherichia coli
41.7
-
 7,8-dihydrofolate mutant linear ANLYF-G7, 15°C, pH 7.0 Escherichia coli
43.7
-
 7,8-dihydrofolate mutant circular ANLYF-G6, 15°C, pH 7.0 Escherichia coli