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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About This Item
CAS Number:
UNSPSC Code:
12352204
eCl@ss:
32160410
EC Number:
232-973-4
NACRES:
NA.54
MDL number:
Specific activity:
30-350 units/mg protein (E1%/280)
Biological source:
Geobacillus stearothermophilus
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Product Name
Thermolysin from Geobacillus stearothermophilus, Type X, lyophilized powder, 30-350 units/mg protein (E1%/280)
biological source
Geobacillus stearothermophilus
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
Quality Level
Related Categories
1 of 4
This Item | P8038 | T7902 | P3910 |
|---|---|---|---|
| specific activity 30-350 units/mg protein (E1%/280) | specific activity ≤0.05 Kunitz units/mg solid RNase, ≤5.0 Kunitz units/mg solid DNase, 7.0-14.0 units/mg solid | specific activity 30-350 units/mg protein (E1%/280) | specific activity - |
| biological source Geobacillus stearothermophilus | biological source - | biological source Geobacillus stearothermophilus | biological source - |
| form lyophilized powder | form lyophilized powder | form powder | form lyophilized powder |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. 2-8°C |
| mol wt 34.6 kDa by amino acid sequence | mol wt 27 kDa | mol wt 34.6 kDa | mol wt 27 kDa |
| shipped in wet ice | shipped in wet ice | shipped in - | shipped in wet ice |
Analysis Note
Contains many extraneous enzymes.
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. [1] It has also been used in a study to investigate the effects of sodium chloride on thermal stability and catalytic activity. [2]
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. [3]
General description
Thermolysin is a protease that has specificity different from other proteases available for sequence investigations. [4]
Other Notes
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.
signalword
Danger
hcodes
Hazard Classifications
Resp. Sens. 1
Storage Class
11 - Combustible Solids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
dust mask type N95 (US), Eyeshields, Faceshields, Gloves
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Yongjin Park et al.
Scientific reports, 12(1), 10935-10935 (2022-06-30)
Long wavelengths that can deeply penetrate into human skin are required to maximize therapeutic effects. Hence, various studies on near-infrared organic light-emitting diodes (NIR OLEDs) have been conducted, and they have been applied in numerous fields. This paper presents a
M Miyanaga et al.
Biotechnology and bioengineering, 46(6), 631-635 (1995-06-20)
N-(benzyloxycarbonyl)-L-aspartyl-L-phenylalanine methyl ester, a precursor of the synthetic sweetener, aspartame, was synthesized from N-(benzyloxycarbonyl)-L-aspartic acid and L-phenylalanine methyl ester with an immobilized thermolysin (EC 3.4.24.4) in the mixed organic solvent system of tert-amyl alcohol and ethyl acetate. A mixed solvent
Friedrich Hans Kleiner et al.
Journal of cell science, 134(9) (2021-09-23)
Cytochrome c6 is a redox carrier in the thylakoid lumen of cyanobacteria and some eukaryotic algae. Although the isofunctional plastocyanin is present in land plants and the green alga Chlamydomonas reinhardtii, these organisms also possess a cytochrome c6-like protein designated
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
The use of thermolysin in amino acid sequence determination.
R P Ambler et al.
The Biochemical journal, 108(5), 893-895 (1968-08-01)
Related Content
QC Methods
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Does the enzyme need to add additional Ca2+ and Zn2+ to be activated?
1 answer-
Helpful?
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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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