Literature summary for 3.4.24.27 extracted from

Ceruso, M.; Howe, N.; Malthouse, J.P.
Mechanism of the binding of Z-L-tryptophan and Z-L-phenylalanine to thermolysin and stromelysin-1 in aqueous solutions (2012), Biochim. Biophys. Acta, 1824, 303-310.

Search for more literature for 3.4.24.27

Inhibitors

Inhibitors	Comment	Organism
2-phenylpropionyl-L-phenylalanine	-	Bacillus sp. (in: Bacteria)
2-phenylpropionyl-Leu-Trp	-	Bacillus sp. (in: Bacteria)
beta-phenylpropionyl-L-phenylalanine	-	Bacillus thermoproteolyticus
additional information	the active site zinc atom is tetrahedrally coordinated when the inhibitors N-benzyloxycarbonyl-tryptophan or N-benzyloxycarbonyl-phenylalanine are bound to thermolysin. pH dependencies of inhibitors N-benzyloxycarbonyl-tryptophan or N-benzyloxycarbonyl-phenylalanine and binding to thermolysin, detailed overview	Bacillus sp. (in: Bacteria)
N-Benzyloxycarbonyl-L-phenylalanine	binding mechanism in aqueous solution, NMR and spectrophotometric analysis, overview. Substitution of Zn2+ by Co2+ decreases the binding affinity of the inhibitor, overview	Bacillus sp. (in: Bacteria)
N-benzyloxycarbonyl-L-tryptophan	binding mechanism in aqueous solution, NMR and spectrophotometric analysis, overview. Substitution of Zn2+ by Co2+ decreases the binding affinity of the inhibitor, overview. With thermolysin, a 1 M concentration of NaSCN produces an 2fold increase in its Ki value from 0.087 mM to 0.19 mM for the inhibitor N-benzyloxycarbonyl-tryptophan	Bacillus sp. (in: Bacteria)
P-Leu-Trp	-	Bacillus thermoproteolyticus
phosphoramidon	-	Bacillus sp. (in: Bacteria)
phosphoramidon	-	Bacillus thermoproteolyticus
Z-L-phenylalanine	enzyme binding structure and kinetics, chemical shift of the carboxylate carbon upon enzyme binding, overview. The carbobenzyloxyl protecting group and not the phenylalanyl phenyl group that is bound in the S1' specificity pocket and the alpha carboxylate group is directly coordinated to the active site zinc atom	Bacillus thermoproteolyticus
Z-L-tryptophan	inhibits full length stromelysin_1-477 and truncated stromelysin_100-264, enzyme binding structure and kinetics, chemical shift of the carboxylate carbon upon enzym ebinding, overview. The tryptophan side chain can bind in the S1 specificity site of stromelysin with the tryptophan alpha carboxylate group coordinated to the active site zinc atom. L-tryptophan binds equally strongly to zinc or cobalt substituted thermolysin	Bacillus thermoproteolyticus

Metals/Ions

Metals/Ions	Comment	Organism
Ca2+	required	Bacillus sp. (in: Bacteria)
Co2+	the active site zinc atom of thermolysin is replaced by cobalt	Bacillus thermoproteolyticus
NaSCN	3fold increase in catalytic activity of thermolysin when the NaSCN concentration is increased to 1 M, but decrease in catalytic activity at higher concentrations of NaSCN	Bacillus sp. (in: Bacteria)
Zn2+	metalloprotease	Bacillus thermoproteolyticus
Zn2+	dependent on, thermolysin is a bacterial zinc metalloproteinase. The active site zinc atom is tetrahedrally coordinated when the inhibitors N-benzyloxycarbonyl-tryptophan or N-benzyloxycarbonyl-phenylalanine are bound to thermolysin	Bacillus sp. (in: Bacteria)

Organism

Organism	UniProt	Comment	Textmining
Bacillus sp. (in: Bacteria)	-	-	-
Bacillus thermoproteolyticus	-	-	-

Source Tissue

Source Tissue	Comment	Organism	Textmining
commercial preparation	-	Bacillus sp. (in: Bacteria)	-
commercial preparation	enzyme crystals	Bacillus thermoproteolyticus	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
9340	-	pH and temperature not specified in the publication	Bacillus thermoproteolyticus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide + H2O	-	Bacillus sp. (in: Bacteria)	N-[3-(2-furyl)acryloyl]-glycine + L-leucine amide	-	?

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	assay at	Bacillus thermoproteolyticus
37	-	assay at	Bacillus sp. (in: Bacteria)

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	7.5	assay at	Bacillus sp. (in: Bacteria)
7	-	-	Bacillus thermoproteolyticus

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism
0.000015	-	2-phenylpropionyl-Leu-Trp	pH 7.2, temperature not specified in the publication	Bacillus sp. (in: Bacteria)
0.000015	-	P-Leu-Trp	pH 7.2, 25°C, enzyme contains Zn2+	Bacillus thermoproteolyticus
0.00003	-	phosphoramidon	pH 7.2, temperature not specified in the publication	Bacillus sp. (in: Bacteria)
0.00003	-	phosphoramidon	pH 7.2, 25°C, enzyme contains Zn2+	Bacillus thermoproteolyticus
0.007	-	N-benzyloxycarbonyl-L-tryptophan	pH 5.0, 37°C, Co2+-bound enzyme	Bacillus sp. (in: Bacteria)
0.007	-	Z-L-tryptophan	pH 5.0, 25°C, enzyme contains Co2+	Bacillus thermoproteolyticus
0.008	-	N-Benzyloxycarbonyl-L-phenylalanine	pH 5.0, 37°C, Co2+-bound enzyme	Bacillus sp. (in: Bacteria)
0.008	-	N-benzyloxycarbonyl-L-tryptophan	pH 5.0, 37°C, Zn2+-bound enzyme	Bacillus sp. (in: Bacteria)
0.008	-	Z-L-phenylalanine	pH 5.0, 25°C, enzyme contains Co2+	Bacillus thermoproteolyticus
0.008	-	Z-L-tryptophan	pH 5.0, 25°C, enzyme contains Zn2+	Bacillus thermoproteolyticus
0.012	-	N-Benzyloxycarbonyl-L-phenylalanine	pH 5.1, 37°C, Zn2+-bound enzyme	Bacillus sp. (in: Bacteria)
0.012	-	Z-L-tryptophan	pH 5.1, 25°C, enzyme contains Zn2+	Bacillus thermoproteolyticus
0.019	-	Z-L-tryptophan	pH 5.0, 25°C, enzyme contains Zn2+, presence of 1 M NaSCN	Bacillus thermoproteolyticus
0.021	-	Z-L-tryptophan	pH 5.0, 25°C, enzyme contains Co2+, presence of 1 M NaSCN	Bacillus thermoproteolyticus
0.087	-	N-benzyloxycarbonyl-L-tryptophan	pH 7.0, 37°C, Zn2+-bound enzyme	Bacillus sp. (in: Bacteria)
0.087	-	Z-L-tryptophan	pH 7.0, 25°C, enzyme contains Zn2+	Bacillus thermoproteolyticus
0.16	-	N-Benzyloxycarbonyl-L-phenylalanine	pH 7.1, 37°C, Co2+-bound enzyme	Bacillus sp. (in: Bacteria)
0.16	-	Z-L-phenylalanine	pH 7.1, 25°C, enzyme contains Co2+	Bacillus thermoproteolyticus
0.19	-	N-benzyloxycarbonyl-L-tryptophan	pH 7.0, 37°C, Zn2+-bound enzyme, in presence of 1 M NaSCN	Bacillus sp. (in: Bacteria)
0.21	-	N-benzyloxycarbonyl-L-tryptophan	pH 7.0, 37°C, Co2+-bound enzyme, in presence of 1 M NaSCN	Bacillus sp. (in: Bacteria)
0.37	-	N-Benzyloxycarbonyl-L-phenylalanine	pH 7.0, 37°C, Zn2+-bound enzyme	Bacillus sp. (in: Bacteria)
0.37	-	Z-L-phenylalanine	pH 7.0, 25°C, enzyme contains Zn2+	Bacillus thermoproteolyticus
1.6	-	2-phenylpropionyl-L-phenylalanine	pH 8.6, temperature not specified in the publication	Bacillus sp. (in: Bacteria)
1.6	-	beta-phenylpropionyl-L-phenylalanine	pH 6.8, 25°C, enzyme contains Zn2+	Bacillus thermoproteolyticus