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P1512

Thermolysin from Geobacillus stearothermophilus

Type X, lyophilized powder, 30-350 units/mg protein (E1%/280)

Synonim(y):

Protease from Geobacillus stearothermophilus, Thermophilic-bacterial protease

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Informacje o tej pozycji

Numer CAS:
UNSPSC Code:
12352204
eCl@ss:
32160410
EC Number:
232-973-4
NACRES:
NA.54
MDL number:
Numer WE:
Specific activity:
30-350 units/mg protein (E1%/280)
Assay:
20—70% (UV)
Biological source:
Geobacillus stearothermophilus
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biological source

Geobacillus stearothermophilus

Quality Segment

type

Type X

assay

20—70% (UV)

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.

Application

Thermolysin has been shown to have a prosequeence that acts as an intramolecular chaperone in vivo. It has also been used in a study to investigate the effects of sodium chloride on thermal stability and catalytic activity.
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.
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.

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.

Analysis Note

Contains many extraneous enzymes.

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).
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Ta pozycja
T7902P2783
description

Type X, lyophilized powder, 30-350 units/mg protein (E1%/280)

description

powder, BioReagent, 30-350 units/mg protein (E1%/280), suitable for cell culture

description

lyophilized powder, ≥3,000 units/mg protein (biuret)

specific activity

30-350 units/mg protein (E1%/280)

specific activity

30-350 units/mg protein (E1%/280)

specific activity

≥3,000 units/mg protein (biuret)

biological source

Geobacillus stearothermophilus

biological source

Geobacillus stearothermophilus

biological source

-

form

lyophilized powder

form

powder

form

lyophilized powder

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

mol wt

34.6 kDa by amino acid sequence

mol wt

34.6 kDa

mol wt

-

shipped in

wet ice

shipped in

-

shipped in

wet ice


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Słowo ostrzegawcze

Danger

Klasa składowania

11 - Combustible Solids

Temperatura zapłonu (°F)

Not applicable

Temperatura zapłonu (°C)

Not applicable

PPE (środki ochrony indywidualnej)

dust mask type N95 (US), Eyeshields, Faceshields, Gloves

Narządy docelowe

Respiratory system

Piktogramy

Health hazardExclamation mark

Kody zagrożeń

Hazard Classifications

Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3

WGK

WGK 1



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Questions

1–2 of 2 Questions  
  1. Does the enzyme need to add additional Ca2+ and Zn2+ to be activated?

    1 answer
    1. 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

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  2. Can this product be dissolved in PBS?

    Can this product be dissolved in PBS?

    1 answer
    1. 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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