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Literature summary for 1.13.11.88 extracted from

  • Lu, X.; Wu, X.; Ma, B.; Xu, Y.
    Enhanced thermostability of Pseudomonas nitroreducens isoeugenol monooxygenase by the combinatorial strategy of surface residue replacement and consensus mutagenesis (2021), Catalysts, 11, 1199 .
No PubMed abstract available
Search for more literature for 1.13.11.88

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Pseudomonas nitroreducens

Protein Variants

Protein Variants Comment Organism
G398A the mutation results in a 1.3fold increase in specific activity Pseudomonas nitroreducens
I352R the mutant with 99.7% activity shows better thermostability than the wild type enzyme (70.4% residual activity after 15 min at 35°C) Pseudomonas nitroreducens
K83R the mutant with 103.9% activity shows better thermostability than the wild type enzyme (84.2% residual activity after 15 min at 35°C) Pseudomonas nitroreducens
K83R/I352R the mutant with 99.2% activity shows better thermostability than the wild type enzyme (87.8% residual activity after 15 min at 35°C) Pseudomonas nitroreducens
K83R/K95R the mutant with 104.8% activity shows better thermostability than the wild type enzyme (92.1% residual activity after 15 min at 35°C) Pseudomonas nitroreducens
K83R/K95R/G398A the mutant with 130.1% activity shows better thermostability than the wild type enzyme (94.1% residual activity after 15 min at 35°C) Pseudomonas nitroreducens
K83R/K95R/I352R the mutant with 114.3% activity shows better thermostability than the wild type enzyme (90.1% residual activity after 15 min at 35°C) Pseudomonas nitroreducens
K83R/K95R/L273F compared with the wild type enzyme, the thermal inactivation half-lives (t1/2) of the mutant at 25°C, 30°C, and 35°C increase 2.9fold, 11.9fold, and 24.7fold, respectively. Simultaneously, it also exhibits a 4.8fold increase in kcat, leading to a 1.2fold increase in catalytic efficiency. The tolerance of the mutant to metal ions is also improved Pseudomonas nitroreducens
K95R the mutant with 106.6% activity shows better thermostability than the wild type enzyme (68% residual activity after 15 min at 35°C) Pseudomonas nitroreducens
L273F the mutant exhibits 1.17fold increase in specific activity compared to the wild type enzyme Pseudomonas nitroreducens

Inhibitors

Inhibitors Comment Organism Structure
Ca2+ 64.6% residual activity at 10 mM Pseudomonas nitroreducens
Co2+ about 80% residual activity at 10 mM Pseudomonas nitroreducens
Fe3+ 21.6% residual activity at 10 mM Pseudomonas nitroreducens
K+ about 90 % residual activity at 10 mM Pseudomonas nitroreducens
Mg2+ about 90 % residual activity at 10 mM Pseudomonas nitroreducens
Mn2+ 69.4% residual activity at 10 mM Pseudomonas nitroreducens
Ni2+ about 45% residual activity at 10 mM Pseudomonas nitroreducens
Vanillin there is an incremental decrease in enzyme activity with the increasing vanillin concentration Pseudomonas nitroreducens
Zn2+ about 50% residual activity at 10 mM Pseudomonas nitroreducens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.1
-
 isoeugenol wild type enzyme, at pH 8.0 and 30°C Pseudomonas nitroreducens
0.3
-
 isoeugenol mutant enzyme L273F, at pH 8.0 and 30°C Pseudomonas nitroreducens
0.4
-
 isoeugenol mutant enzyme K83R/K95R/L273F, at pH 8.0 and 30°C Pseudomonas nitroreducens
0.4
-
 isoeugenol mutant enzyme K95R, at pH 8.0 and 30°C Pseudomonas nitroreducens
0.5
-
 isoeugenol mutant enzyme K83R/K95R, at pH 8.0 and 30°C Pseudomonas nitroreducens
0.6
-
 isoeugenol mutant enzyme K83R, at pH 8.0 and 30°C Pseudomonas nitroreducens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
isoeugenol + O2 Pseudomonas nitroreducens
-
 vanillin + acetaldehyde
-
 ?
isoeugenol + O2 Pseudomonas nitroreducens Jin1
-
 vanillin + acetaldehyde
-
 ?

Organism

Organism UniProt Comment Textmining
Pseudomonas nitroreducens
-
-
-
Pseudomonas nitroreducens Jin1
-
-
-

Purification (Commentary)

Purification (Comment) Organism
BeaverBeads column chromatography Pseudomonas nitroreducens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
isoeugenol + O2
-
 Pseudomonas nitroreducens vanillin + acetaldehyde
-
 ?
isoeugenol + O2
-
 Pseudomonas nitroreducens Jin1 vanillin + acetaldehyde
-
 ?

Subunits

Subunits Comment Organism
? x * 50000, SDS-PAGE Pseudomonas nitroreducens

Synonyms

Synonyms Comment Organism
iem
-
 Pseudomonas nitroreducens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
-
 Pseudomonas nitroreducens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
25 35 the wild type enzyme shows 53.2% residual activity after 15 min at 35°C. The enzyme has half-lives of 967.2, 49.8 and 7.8 min at 25, 30, and 35°C, respectively Pseudomonas nitroreducens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.1
-
 isoeugenol wild type enzyme, at pH 8.0 and 30°C Pseudomonas nitroreducens
5.5
-
 isoeugenol mutant enzyme K83R, at pH 8.0 and 30°C Pseudomonas nitroreducens
6.1
-
 isoeugenol mutant enzyme K95R, at pH 8.0 and 30°C Pseudomonas nitroreducens
6.9
-
 isoeugenol mutant enzyme L273F, at pH 8.0 and 30°C Pseudomonas nitroreducens
10.1
-
 isoeugenol mutant enzyme K83R/K95R/L273F, at pH 8.0 and 30°C Pseudomonas nitroreducens
10.5
-
 isoeugenol mutant enzyme K83R/K95R, at pH 8.0 and 30°C Pseudomonas nitroreducens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
-
 Pseudomonas nitroreducens

pH Range

pH Minimum pH Maximum Comment Organism
8 9.5 the wild type enzyme remains 70% relative activity in a pH range from 8.0 to 9.5 Pseudomonas nitroreducens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.3
-
 isoeugenol mutant enzyme K95R, at pH 8.0 and 30°C Pseudomonas nitroreducens
5.5
-
 isoeugenol mutant enzyme L273F, at pH 8.0 and 30°C Pseudomonas nitroreducens
6
-
 isoeugenol mutant enzyme K83R/K95R, at pH 8.0 and 30°C Pseudomonas nitroreducens
6.1
-
 isoeugenol mutant enzyme K83R, at pH 8.0 and 30°C Pseudomonas nitroreducens
7
-
 isoeugenol mutant enzyme K83R/K95R/L273F, at pH 8.0 and 30°C Pseudomonas nitroreducens
21
-
 isoeugenol wild type enzyme, at pH 8.0 and 30°C Pseudomonas nitroreducens