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Literature summary extracted from

  • Toogood, H.S.; Hollingsworth, E.J.; Brown, R.C.; Taylor, I.N.; Taylor, S.J.; McCague, R.; Littlechild, J.A.
    A thermostable L-aminoacylase from Thermococcus litoralis: cloning, overexpression, characterization, and applications in biotransformations (2002), Extremophiles, 6, 111-122.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.5.1.14 mono-benzyl malonate 25 mM, activation to 109% of control Thermococcus litoralis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.14 overexpressed in Escherichia coli Thermococcus litoralis
3.5.1.14 overexpressed in Escherichia coli, modification of N-terminal residues of the recombinant enzyme to enhance expression in Escherichia coli using the vector pUCcer11. The recombinant enzyme has an insertion of four amino acids (Glu-Pro-Leu-Phe) between the N-terminal methionine and the second residue lysine of the wild type Thermococcus litoralis

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.1.14 1,10-phenanthroline 1 mM, 54% inhibition Thermococcus litoralis
3.5.1.14 4-hydroxymercuribenzoate 1 mM, 67% inhibition Thermococcus litoralis
3.5.1.14 Ba2+ 0.01 mM, 17% inhibition Thermococcus litoralis
3.5.1.14 benzamidine 0.1 mM, 7% inhibition Thermococcus litoralis
3.5.1.14 Ca2+ 0.01 mM, 45% inhibition Thermococcus litoralis
3.5.1.14 Cs+ 0.01 mM, 47% inhibition Thermococcus litoralis
3.5.1.14 Cu2+ 0.01 mM, 95% inhibition Thermococcus litoralis
3.5.1.14 dithiothreitol 1 mM, 22% inhibition; 1 mM, 42% inhibition Thermococcus litoralis
3.5.1.14 EDTA 1 mM, 45% inhibition Thermococcus litoralis
3.5.1.14 Fe2+ 0.01 mM, 33% inhibition Thermococcus litoralis
3.5.1.14 Fe3+ 0.01 mM, 99% inhibition Thermococcus litoralis
3.5.1.14 iodoacetate 1 mM, 1.1fold activation Thermococcus litoralis
3.5.1.14 Isopropanol 10% Thermococcus litoralis
3.5.1.14 K+ 0.01 mM, 24% inhibition Thermococcus litoralis
3.5.1.14 La3+ 0.01 mM, 17% inhibition Thermococcus litoralis
3.5.1.14 Li+ 0.01 mM, 51% inhibition Thermococcus litoralis
3.5.1.14 Mg2+ 0.01 mM, 8% inhibition Thermococcus litoralis
3.5.1.14 Mn2+ 0.01 mM, 54% inhibition Thermococcus litoralis
3.5.1.14 mono-tert-butyl malonate 25 mM, 14% inhibition Thermococcus litoralis
3.5.1.14 N-(4-toluenesulfonyl)-DL-methionine 25 mM, 14% inhibition Thermococcus litoralis
3.5.1.14 N-(p-toluenesulfonyl)-DL-Met 25 mM, 14% inhibition Thermococcus litoralis
3.5.1.14 Na+ 0.01 mM, 19% inhibition Thermococcus litoralis
3.5.1.14 Ni2+ 0.01 mM, 27% inhibition Thermococcus litoralis
3.5.1.14 phenylmethylsulfonyl fluoride 1 mM, 22% inhibition Thermococcus litoralis
3.5.1.14 Sn4+ 0.01 mM, 99% inhibition Thermococcus litoralis
3.5.1.14 tosyl-L-lysine chloromethyl ketone 25 mM, 7% inhibition Thermococcus litoralis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.1.14 Co2+ 0.01 mM, 1.1fold activation Thermococcus litoralis
3.5.1.14 Co2+ 0.01 mM, 1.1fold activation. 0.1 mM, 1.3fold activation Thermococcus litoralis
3.5.1.14 Zn2+ 0.01 mM or 0.1 mM, 1.3fold activation Thermococcus litoralis
3.5.1.14 Zn2+ 0.01 mM, 1.3fold activation Thermococcus litoralis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.1.14 43000
-
 4 * 43000, SDS-PAGE Thermococcus litoralis
3.5.1.14 43000
-
 4 * 43000, SDS-PAGE, sedimentation studies Thermococcus litoralis
3.5.1.14 43814
-
 4 * 43814, calculated from sequence Thermococcus litoralis
3.5.1.14 43815
-
 4 * 43815, calculated from sequence Thermococcus litoralis
3.5.1.14 170000
-
 gel filtration Thermococcus litoralis

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.1 Thermococcus litoralis
-
-
-
3.5.1.14 Thermococcus litoralis
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.1 4-nitrophenyl laurate + H2O
-
 Thermococcus litoralis 4-nitrophenol + laurate
-
 ?
3.1.1.1 4-nitrophenyl palmitate + H2O
-
 Thermococcus litoralis 4-nitrophenol + palmitate
-
 ?
3.1.1.1 N-acetyl-L-phenylalanine methyl ester + H2O
-
 Thermococcus litoralis N-acetyl-L-phenylalanine + methanol
-
 ?
3.5.1.14 additional information the enzyme is most specific for substrates containing N-benzoyl- or N-chloroacetyl-amino acids, preferring substrates containing hydrophobic, uncharged, or weakly charged amino acids such as phenylalanine, methionine, and cysteine. It contains esterase activity with 4-nitrophenyl-palmitate and 4-nitrophenyllaurate, but not Phe-ethyl ester at 85°C Thermococcus litoralis ?
-
 ?
3.5.1.14 additional information the enzyme is most specific for substrates containing N-benzoyl- or N-chloroacetyl-amino acids, preferring substrates containing hydrophobic, uncharged, or weakly charged amino acids such as phenylalanine, methionine, and cysteine. No activity with N-benzoyl-D-phenylalanine, N-benzyloxycarbonyl-D-phenylalanine, tert-butyloxycarbonyl-D-phenylalanine, N-acetyl-D-tryptophan Thermococcus litoralis ?
-
 ?
3.5.1.14 N-acetyl-L-cysteine + H2O
-
 Thermococcus litoralis acetate + L-cysteine
-
 ?
3.5.1.14 N-acetyl-L-methionine + H2O
-
 Thermococcus litoralis acetate + L-methionine
-
 ?
3.5.1.14 N-acetyl-L-methionine + H2O 40.5% of the activity with N-acetyl-L-phenylalanine Thermococcus litoralis acetate + L-methionine
-
 ?
3.5.1.14 N-acetyl-L-Phe methyl ester + H2O the enzyme also shows esterase activity with N-acetyl-L-Phe methyl ester Thermococcus litoralis acetate + L-Phe methyl ester
-
 ?
3.5.1.14 N-acetyl-L-phenylalanine + H2O no activity with N-acetyl-D-phenylalanine Thermococcus litoralis acetate + L-phenylalanine
-
 ?
3.5.1.14 N-acetyl-L-phenylalanine methyl ester + H2O
-
 Thermococcus litoralis acetate + L-phenylalanine methyl ester
-
 ?
3.5.1.14 N-acetyl-L-tryptophan + H2O
-
 Thermococcus litoralis acetate + L-tryptophan
-
 ?
3.5.1.14 N-benzoyl-L-phenylalanine + H2O
-
 Thermococcus litoralis benzoate + L-phenylalanine
-
 ?
3.5.1.14 N-benzoyl-L-phenylalanine + H2O 735.7% of the activity with N-acetyl-L-phenylalanine Thermococcus litoralis benzoate + L-phenylalanine
-
 ?
3.5.1.14 N-benzyloxycarbonyl-L-Phe-Gly + H2O
-
 Thermococcus litoralis benzyl hydrogencarbonate + L-Phe-Gly
-
 ?
3.5.1.14 N-benzyloxycarbonyl-L-phenylalanine + H2O
-
 Thermococcus litoralis benzyl hydrogencarbonate + L-phenylalanine
-
 ?
3.5.1.14 N-benzyloxycarbonyl-L-phenylalanine + H2O 33.9% of the activity with N-acetyl-L-phenylalanine. no activity with N-benzyloxycarbonyl-D-phenylalanine Thermococcus litoralis benzyloxycarbonate + L-phenylalanine
-
 ?
3.5.1.14 N-benzyloxycarbonyl-L-valine + H2O
-
 Thermococcus litoralis benzyl hydrogencarbonate + L-valine
-
 ?
3.5.1.14 N-chloroacetyl-L-phenylalanine + H2O
-
 Thermococcus litoralis chloroacetate + L-phenylalanine
-
 ?
3.5.1.14 N-tert-butyloxycarbonyl-L-phenylalanine + H2O
-
 Thermococcus litoralis tert-butyloxycarboxylate + L-phenylalanine
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.1.14 homotetramer 4 * 43000, SDS-PAGE Thermococcus litoralis
3.5.1.14 homotetramer 4 * 43000, SDS-PAGE, sedimentation studies Thermococcus litoralis
3.5.1.14 homotetramer 4 * 43814, calculated from sequence Thermococcus litoralis
3.5.1.14 homotetramer 4 * 43815, calculated from sequence Thermococcus litoralis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.1.14 80
-
 assay at Thermococcus litoralis
3.5.1.14 85
-
 at pH 8.0 Thermococcus litoralis
3.5.1.14 85
-
 1 h, Tris-HCl at pH 8.0 Thermococcus litoralis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.5.1.14 70
-
 half-life: 25 h Thermococcus litoralis
3.5.1.14 85
-
 half-life: 1.7 h Thermococcus litoralis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.1.14 7
-
 assay at Thermococcus litoralis
3.5.1.14 7.5
-
 with Mes buffer Thermococcus litoralis
3.5.1.14 8
-
 assay at Thermococcus litoralis
3.5.1.14 8
-
 with Tris-HCl or phosphate buffers Thermococcus litoralis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.5.1.14 6.5 9.5 at least 70% of the activity is present between pH 6.5 and 9.5 Thermococcus litoralis