The recommended assay buffer includes calcium: 10 mM sodium acetate buffer with 5 mM calcium acetate, pH 7.5 at 37°C. Prepare 100 ml in deionized water using sodium acetate trihydrate and calcium acetate. Adjust the pH to 7.5 at 37°C with 0.1 M acetic acid or 0.1 M NaOH. Additional information can be found at: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/390/551/p1512enz.pdf
P1512
Thermolysin from Geobacillus stearothermophilus
Type X, lyophilized powder, 30-350 units/mg protein (E1%/280)
Synonym(s):
Protease from Geobacillus stearothermophilus, Thermophilic-bacterial protease
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25 MG
MXP 2,526.00
100 MG
MXP 5,969.00
250 MG
MXP 11,604.00
1 G
MXP 29,637.00
MXP 2,526.00
Estimated to ship onApril 12, 2025
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25 MG
MXP 2,526.00
100 MG
MXP 5,969.00
250 MG
MXP 11,604.00
1 G
MXP 29,637.00
About This Item
MXP 2,526.00
Estimated to ship onApril 12, 2025
Recommended Products
biological source
Geobacillus stearothermophilus
Quality Level
type
Type X
form
lyophilized powder
specific activity
30-350 units/mg protein (E1%/280)
mol wt
34.6 kDa by amino acid sequence
purified by
crystallization
shipped in
wet ice
storage temp.
−20°C
General description
Thermolysin is a protease that has specificity different from other proteases available for sequence investigations. [1]
Application
A thermostable (thermophilic) extracellular metalloendopeptidase containing four calcium ions. Cofactors are zinc and calcium. Hydrolyzes protein bonds on the N-terminal side of hydrophobic amino acid residues. The pH optimum is 8.0 and the optimal temperature for activity is 70 °C. Considerably stable from pH 5 to 9.5. Thermolysin has a low cleavage specificity, therefore, it produces a number of short fragments that are suitable for sequencing. Preferential cleavage: X-cleavage-Y-Z where X=any amino acid; Y=Leu, Phe, Ile, Val, Met, Ala and Z is any amino acid other than Pro. Cleavage N-terminal to Leu is preferred over cleavage of N-terminal to Phe which is preferred over the others. Often used to do limited proteolysis for peptide mapping and studies of protein structure and conformational changes.
Thermolysin has been shown to have a prosequeence that acts as an intramolecular chaperone in vivo. [2] It has also been used in a study to investigate the effects of sodium chloride on thermal stability and catalytic activity. [3]
Thermolysin is also commonly used for the commercial synthesis of N-(benzyloxycarbonyl)-L-aspartyl-L-phenylalanine methyl ester, the precursor for the artificial sweetener aspartame. [4]
Quality
Contains many extraneous enzymes.
Unit Definition
One unit will hydrolyze casein to produce color equivalent to 1.0 μmole (181 μg) of tyrosine per min at pH 7.5 at 37 °C (color by Folin-Ciocalteu reagent).
Physical form
lyophilized powder containing calcium and sodium acetate buffer salts
Preparation Note
The pH optimum is 8.0 and the optimal temperature for activity is 70 °C. Considerably stable from pH 5 to 9.5. Thermolysin has a low cleavage specificity, therefore, it produces a number of short fragments that are suitable for sequencing. Preferential cleavage: X-cleavage-Y-Z where X=any amino acid; Y=Leu, Phe, Ile, Val, Met, Ala and Z is any amino acid other than Pro. Cleavage N-terminal to Leu is preferred over cleavage of N-terminal to Phe which is preferred over the others.
Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Resp. Sens. 1
Storage Class Code
11 - Combustible Solids
WGK
WGK 3
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Personal Protective Equipment
dust mask type N95 (US), Eyeshields, Gloves
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Customers Also Viewed
Chiwook Park et al.
Nature methods, 2(3), 207-212 (2005-03-23)
Thermodynamic stability is fundamental to the biology of proteins. Information on protein stability is essential for studying protein structure and folding and can also be used indirectly to monitor protein-ligand or protein-protein interactions. While clearly valuable, the experimental determination of
C Marie-Claire et al.
Journal of molecular biology, 285(5), 1911-1915 (1999-02-02)
The zinc metalloendopeptidase, thermolysin (EC 3.4.24.27) produced by Bacillus thermoproteolyticus serves as a model of important physiological enzymes such as neprilysin, angiotensin converting enzyme and endothelin converting enzyme. Thermolysin is synthesised as a pre-proenzyme, with an N-terminal prosequence of 204
Maria Fernanda Lay Mendoza et al.
Viruses, 12(12) (2020-12-23)
Viral entry is the first stage in the virus replication cycle and, for enveloped viruses, is mediated by virally encoded glycoproteins. Viral glycoproteins have different receptor affinities and triggering mechanisms. We employed vesicular stomatitis virus (VSV), a BSL-2 enveloped virus
The use of thermolysin in amino acid sequence determination.
R P Ambler et al.
The Biochemical journal, 108(5), 893-895 (1968-08-01)
K Inouye et al.
Biochimica et biophysica acta, 1388(1), 209-214 (1998-10-17)
Thermolysin, a thermophilic metalloproteinase, is markedly activated in the presence of high concentrations (1-5 M) of neutral salts. The activity increases in an exponential fashion with increasing salt concentration, and is enhanced 13-15 times with 4 M NaCl at pH
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Does the enzyme need to add additional Ca2+ and Zn2+ to be activated?
1 answer-
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Can this product be dissolved in PBS?
Can this product be dissolved in PBS?
1 answer-
There is no specific information available on the dissolution of thermolysin at a 50 mM concentration in PBS. PBS typically contains between 138 to 150 mM sodium chloride, which may or may not lower the overall solubility of the enzyme in PBS. If PBS is used, the user will need to determine if the enzyme fully dissolves in the solution or not.
For the complete enzymatic procedure, you can find the link here: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/390/551/p1512enz.pdf
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