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)
Sinonimo/i:
Protease from Geobacillus stearothermophilus, Thermophilic-bacterial protease
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About This Item
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Origine biologica
Geobacillus stearothermophilus
Tipo
Type X
Stato
lyophilized powder
Attività specifica
30-350 units/mg protein (E1%/280)
PM
34.6 kDa by amino acid sequence
Purificato mediante
crystallization
Condizioni di spedizione
wet ice
Temperatura di conservazione
−20°C
Categorie correlate
Descrizione generale
Thermolysin is a protease that has specificity different from other proteases available for sequence investigations. [1]
Applicazioni
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]
Qualità
Contains many extraneous enzymes.
Definizione di unità
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).
Stato fisico
lyophilized powder containing calcium and sodium acetate buffer salts
Nota sulla preparazione
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.
Avvertenze
Danger
Indicazioni di pericolo
Consigli di prudenza
Classi di pericolo
Resp. Sens. 1
Codice della classe di stoccaggio
11 - Combustible Solids
Classe di pericolosità dell'acqua (WGK)
WGK 3
Punto d’infiammabilità (°F)
Not applicable
Punto d’infiammabilità (°C)
Not applicable
Dispositivi di protezione individuale
dust mask type N95 (US), Eyeshields, Faceshields, Gloves
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I clienti hanno visto anche
The use of thermolysin in amino acid sequence determination.
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The Biochemical journal, 108(5), 893-895 (1968-08-01)
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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
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
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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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