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Literature summary extracted from

  • Chen, B.; Fan, L.; Xu, J.; Zhao, J.; Zhang, X.; Ouyang, L.
    Biocatalytic properties of a recombinant Fusarium proliferatum lactonase with significantly enhanced production by optimal expression in Escherichia coli (2010), Appl. Biochem. Biotechnol., 162, 744-756.
    View publication on PubMed

Application

EC Number Application Comment Organism
3.1.1.25 synthesis the recombinant enzyme is a highly efficient biocatalyst for asymmetric synthesis of chiral compounds Fusarium proliferatum

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.1.25 expressed in Escherichia coli JM109(DE3) cells Fusarium proliferatum

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.1.1.25 Ag+ 33.2% residual activity at 1 mM Fusarium proliferatum
3.1.1.25 Al3+ 22.4% residual activity at 1 mM Fusarium proliferatum
3.1.1.25 Co2+ 10.7% residual activity at 1 mM Fusarium proliferatum
3.1.1.25 Cu+ 13.6% residual activity at 1 mM Fusarium proliferatum
3.1.1.25 Cu2+ 11.2% residual activity at 1 mM Fusarium proliferatum
3.1.1.25 Fe2+ 15.6% residual activity at 1 mM Fusarium proliferatum
3.1.1.25 Fe3+ 3.3% residual activity at 1 mM Fusarium proliferatum
3.1.1.25 Hg2+ 2.1% residual activity at 1 mM Fusarium proliferatum
3.1.1.25 Li+ 24.7% residual activity at 1 mM Fusarium proliferatum
3.1.1.25 Mg2+ 32.4% residual activity at 1 mM Fusarium proliferatum
3.1.1.25 Mn2+ 22.7% residual activity at 1 mM Fusarium proliferatum
3.1.1.25 additional information EDTA does not exhibit significant effect on enzyme activity Fusarium proliferatum
3.1.1.25 Ni2+ 55.4% residual activity at 1 mM Fusarium proliferatum
3.1.1.25 Pb2+ 13.9% residual activity at 1 mM Fusarium proliferatum
3.1.1.25 Zn2+ 14.3% residual activity at 1 mM Fusarium proliferatum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.1.25 121
-
 DL-pantoyllactone recombinant enzyme, at pH 6.7-6.9, 30°C Fusarium proliferatum
3.1.1.25 290.2
-
 2-hydroxybutyrolactone recombinant enzyme, at pH 6.7-6.9, 30°C Fusarium proliferatum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.1.1.25 Ca2+ 7.4% activation at 1 mM. No significant effect of Ca2+ on the stability of recombinant enzyme is observed when incubated together for 60 h at 30°C or 4°C Fusarium proliferatum
3.1.1.25 additional information the enzyme is not affected by Na+ and K+ Fusarium proliferatum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.1.1.25 50000
-
 x * 50000, recombinant protein, SDS-PAGE Fusarium proliferatum

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
3.1.1.25 Acetone 49% residual activity at 10% (v/v), when the concentration of methanol is increased up to 20% (v/v), up to 30% of the initial activity is retained Fusarium proliferatum
3.1.1.25 acetonitrile 55% residual activity at 10% (v/v), when the concentration of methanol is increased up to 20% (v/v), up to 30% of the initial activity is retained Fusarium proliferatum
3.1.1.25 dimethylformamide 51% residual activity at 10% (v/v) Fusarium proliferatum
3.1.1.25 Ethanol 55% residual activity at 10% (v/v) Fusarium proliferatum
3.1.1.25 Methanol 81% residual activity at 10% (v/v), when the concentration of methanol is increased up to 20% (v/v), 60.5% of the initial activity is retained Fusarium proliferatum

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.25 Fusarium proliferatum B2LYJ5
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.1.25 Ni-NTA column chromatography Fusarium proliferatum

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.1.1.25 341.2
-
 recombinant enzyme after 14.4fold purification, using DL-pantoyl lactone as substrate, at pH 6.7-6.9, 30°C Fusarium proliferatum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.25 2-bromobutyrolactone + H2O less than 1% activity compared to DL-pantoyl lactone Fusarium proliferatum ?
-
 ?
3.1.1.25 2-hydroxybutyrolactone + H2O 2450% activity compared to DL-pantoyl lactone Fusarium proliferatum ?
-
 ?
3.1.1.25 4-butyrolactone + H2O less than 1% activity compared to DL-pantoyl lactone Fusarium proliferatum ?
-
 ?
3.1.1.25 4-phenyl-2-hydroxyl-4-butyrolactone + H2O 22% activity compared to DL-pantoyl lactone Fusarium proliferatum ?
-
 ?
3.1.1.25 DL-pantoyllactone + H2O 100% activity Fusarium proliferatum pantoic acid
-
 ?
3.1.1.25 ethyl acetate + H2O less than 1% activity compared to DL-pantoyl lactone Fusarium proliferatum ?
-
 ?
3.1.1.25 methyl 2-chloroacetate + H2O 11.6% activity compared to DL-pantoyl lactone Fusarium proliferatum ?
-
 ?
3.1.1.25 methyl 2-chloropropanoate + H2O 2.3% activity compared to DL-pantoyl lactone Fusarium proliferatum ?
-
 ?
3.1.1.25 methyl lactate + H2O less than 1% activity compared to DL-pantoyl lactone Fusarium proliferatum ?
-
 ?
3.1.1.25 methyl mandelate + H2O less than 1% activity compared to DL-pantoyl lactone Fusarium proliferatum ?
-
 ?
3.1.1.25 additional information no activity with 4-nitrophenyl acetate and 4-nitrophenyl butyrate Fusarium proliferatum ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.1.1.25 ? x * 50000, recombinant protein, SDS-PAGE Fusarium proliferatum

Synonyms

EC Number Synonyms Comment Organism
3.1.1.25 levo-lactonase
-
 Fusarium proliferatum

Expression

EC Number Organism Comment Expression
3.1.1.25 Fusarium proliferatum by increasing the biomass concentration and lowering the inducer (isopropyl-beta-D-thiogalactoside) concentration and induction temperature, the recombinant lactonase production in Escherichia coli is significantly enhanced (20times higher than that of wild type strain Fusarium proliferatum ECU2002) up