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)
Synonyme(s) :
Protease from Geobacillus stearothermophilus, Thermophilic-bacterial protease
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About This Item
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Produits recommandés
Source biologique
Geobacillus stearothermophilus
Niveau de qualité
Type
Type X
Forme
lyophilized powder
Activité spécifique
30-350 units/mg protein (E1%/280)
Poids mol.
34.6 kDa by amino acid sequence
Produit purifié par
crystallization
Conditions d'expédition
wet ice
Température de stockage
−20°C
Description générale
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]
Qualité
Contains many extraneous enzymes.
Définition de l'unité
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).
Forme physique
lyophilized powder containing calcium and sodium acetate buffer salts
Notes préparatoires
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.
Mention d'avertissement
Danger
Mentions de danger
Conseils de prudence
Classification des risques
Resp. Sens. 1
Code de la classe de stockage
11 - Combustible Solids
Classe de danger pour l'eau (WGK)
WGK 3
Point d'éclair (°F)
Not applicable
Point d'éclair (°C)
Not applicable
Équipement de protection individuelle
dust mask type N95 (US), Eyeshields, Faceshields, Gloves
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Les clients ont également consulté
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
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
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
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
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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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