Literature summary for 3.1.1.25 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

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

Cloned(Commentary)

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

Inhibitors

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

KM Value [mM]

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

Metals/Ions

Metals/Ions Comment Organism Structure
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
additional information the enzyme is not affected by Na+ and K+ Fusarium proliferatum

Molecular Weight [Da]

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

Organic Solvent Stability

Organic Solvent Comment Organism
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
Ethanol 55% residual activity at 10% (v/v) Fusarium proliferatum
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
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
dimethylformamide 51% residual activity at 10% (v/v) Fusarium proliferatum

Organism

Organism UniProt Comment Textmining
Fusarium proliferatum B2LYJ5
-
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
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)

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

Subunits

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

Synonyms

Synonyms Comment Organism
levo-lactonase
-
Fusarium proliferatum

Expression

Organism Comment Expression
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