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Literature summary 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

Cloned(Commentary)

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

Protein Variants

EC Number Protein Variants Comment Organism
1.13.11.88 G398A the mutation results in a 1.3fold increase in specific activity Pseudomonas nitroreducens
1.13.11.88 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
1.13.11.88 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
1.13.11.88 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
1.13.11.88 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
1.13.11.88 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
1.13.11.88 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
1.13.11.88 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
1.13.11.88 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
1.13.11.88 L273F the mutant exhibits 1.17fold increase in specific activity compared to the wild type enzyme Pseudomonas nitroreducens

Inhibitors

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.13.11.88 0.1
-
 isoeugenol wild type enzyme, at pH 8.0 and 30°C Pseudomonas nitroreducens
1.13.11.88 0.3
-
 isoeugenol mutant enzyme L273F, at pH 8.0 and 30°C Pseudomonas nitroreducens
1.13.11.88 0.4
-
 isoeugenol mutant enzyme K83R/K95R/L273F, at pH 8.0 and 30°C Pseudomonas nitroreducens
1.13.11.88 0.4
-
 isoeugenol mutant enzyme K95R, at pH 8.0 and 30°C Pseudomonas nitroreducens
1.13.11.88 0.5
-
 isoeugenol mutant enzyme K83R/K95R, at pH 8.0 and 30°C Pseudomonas nitroreducens
1.13.11.88 0.6
-
 isoeugenol mutant enzyme K83R, at pH 8.0 and 30°C Pseudomonas nitroreducens

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
1.13.11.88 Pseudomonas nitroreducens
-
-
-
1.13.11.88 Pseudomonas nitroreducens Jin1
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.13.11.88 BeaverBeads column chromatography Pseudomonas nitroreducens

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

EC Number Synonyms Comment Organism
1.13.11.88 iem
-
 Pseudomonas nitroreducens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.13.11.88 30
-
-
 Pseudomonas nitroreducens

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.13.11.88 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]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.13.11.88 2.1
-
 isoeugenol wild type enzyme, at pH 8.0 and 30°C Pseudomonas nitroreducens
1.13.11.88 5.5
-
 isoeugenol mutant enzyme K83R, at pH 8.0 and 30°C Pseudomonas nitroreducens
1.13.11.88 6.1
-
 isoeugenol mutant enzyme K95R, at pH 8.0 and 30°C Pseudomonas nitroreducens
1.13.11.88 6.9
-
 isoeugenol mutant enzyme L273F, at pH 8.0 and 30°C Pseudomonas nitroreducens
1.13.11.88 10.1
-
 isoeugenol mutant enzyme K83R/K95R/L273F, at pH 8.0 and 30°C Pseudomonas nitroreducens
1.13.11.88 10.5
-
 isoeugenol mutant enzyme K83R/K95R, at pH 8.0 and 30°C Pseudomonas nitroreducens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.13.11.88 9
-
-
 Pseudomonas nitroreducens

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.13.11.88 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.13.11.88 5.3
-
 isoeugenol mutant enzyme K95R, at pH 8.0 and 30°C Pseudomonas nitroreducens
1.13.11.88 5.5
-
 isoeugenol mutant enzyme L273F, at pH 8.0 and 30°C Pseudomonas nitroreducens
1.13.11.88 6
-
 isoeugenol mutant enzyme K83R/K95R, at pH 8.0 and 30°C Pseudomonas nitroreducens
1.13.11.88 6.1
-
 isoeugenol mutant enzyme K83R, at pH 8.0 and 30°C Pseudomonas nitroreducens
1.13.11.88 7
-
 isoeugenol mutant enzyme K83R/K95R/L273F, at pH 8.0 and 30°C Pseudomonas nitroreducens
1.13.11.88 21
-
 isoeugenol wild type enzyme, at pH 8.0 and 30°C Pseudomonas nitroreducens