Thermo Scientific Lysozyme is an enzyme characterized by the ability to break down the bacterial cell wall to improve protein or nucleic acid extraction efficiency.
Lysozyme is an enzyme used to break down bacterial cell walls to improve protein or nucleic acid extraction efficiency. Lysozymes (muramidases) are a family of enzymes with antimicrobial activity characterized by the ability to damage the cell wall of bacteria. The enzyme acts by catalyzing the hydrolysis of 1,4-beta-linkages between N-acetylmuramic acid and N-acetyl-D-glucosamine residues in peptidoglycans and between the N-acetyl-D-glucosamine residues in chitodextrins. Although hen egg white lysozyme is most effective for the lysis of gram-positive bacteria, it also facilitates the lysis of gram-negative bacteria such as Salmonella and Shigella. The lysis of E. coli is especially improved by the addition of both lysozyme and a nucleases such as DNase I.
- Bacterial cell wall lytic enzyme that improves inclusion body protein purification
- Compatible with Thermo Scientific Pierce Cell Lysis Reagents
- Available in two convenient formats: lyophilized powder for large volume applications (Part No. 89833) and convenient glycerol freezer stock (Part No. 90082)
|Thermo Scientific Lysozyme improves protein extraction efficiency. To demonstrate the benefit of using lysozyme and DNase I to process bacterial extracts, the extractions of two different-sized, over-expressed proteins were compared. Cell pellets from 50mL culture of E. coli BL-21 over-expressing green fluorescent protein (GFP) or GST-Ral Binding Protein (GST-RalBP) were lysed using B-PER Reagent with and with out lysozyme and DNase I. The soluble fractions were separated from the pellets, and the two fractions were analyzed by SDS-PAGE. Although GFP (32 kDa) was extracted equally well in the absence or presence of enzyme, GST-RalBP (75 kDa) required lysozyme and DNase I to be efficiently solubilized.
Description and Properties of Lysozyme:
Lysozyme occurs naturally in plant and animal tissues and in secretions such as tears, saliva and mucus; it is especially abundant in egg whites, which is the primary source for commercial supplies. Hen egg white lysozyme (chick-type or c) was the first enzyme to have its 2-angstrom crystal structure resolved and has been extensively studied. Lysozyme is widely used in bacterial protein extraction. For inclusion body purification, lysozyme is added to digest cell debris and release the inclusion bodies. The enzyme is easily eliminated with other proteins during fusion tag-specific affinity purification of the target recombinant protein in forming the inclusion body.
- Enzymatic function: Catalyzes the hydrolysis of 1,4-β-linkages between N-acetylmuramic acid and N-acetyl-D-glucosamine residues in peptidoglycans and between N-acetyl-D-glucosamine residues in chitodextrins
- Molecular weight: 14.388 kDa
- Extinction coefficient: ~26.4 at 280nm
- Isoelectric point: pH 11.0
- Unit definition: 1 unit is that amount of enzyme needed to catalyze a decrease in absorbance at 450nm of 0.001/min at 25 degrees at pH 6.24 in a 1cm cuvette due to lysis using a ~0.25mg/mL suspension of Micrococcus luteus.
Specifications (Part No. 89833):
- Visual: White, lyophilized powder
- Activity: Approx. 20,000 units/mg dry weight
- Solubility: Must be readily soluble at 5mg/mL in 0.1M KH2PO4 to give a clear, colorless solution.
Specifications (Part No. 90082):
- Visual: Clear and colorless, free of insoluble material
- pH: 4.7 ±0.2
- Activity: ≥50 units/mL at 5µg/mL
- Alderton, G., Ward, W., and Febold, H. (1945). Isolation of lysozyme from egg white. J. Biol. Chem. 157:43-58.
- Aune, K., and Tanford, C. (1969). Thermodynamics of the denaturation of lysozyme by guanidine hydrochloride. I. Dependence on pH at 25 degrees. Biochem. 8(11): 4579-85.
- Blake, C.C., et al. (1965). Structure of hen egg-white lysozyme: a three-dimensional Fourier synthesis at 2 angstrom resolution. Nature. 206(986): 757-61.
- Hara, S., and Matsushima, Y. (1967). Studies on the substrate specificity of egg white lysozyme. I. The N-acyl substituent in the substrate mucopolysaccharides. J. Biochem. 62: 118-25.
- Hara, S., and Matsushima, Y. (1972). Studies on the substrate specificity of egg white lysozyme. IV. A comparative study of the substrate specificities of lysozymes from different sources. J. Biochem. 72: 993-1000.
- Shinitzky, M., et al. (1966). Inhibition of lysozyme by imidazole and indole derivatives. Arch. Biochem. Biophys. 116: 332-43.
- Smékal, F. (1973). Lysis of lyophilized Eschericial coli cell with egg-white lysozyme without ethylenediaminetetraacetic acid. Folia Microbiologica. 18: 146-8.
- Smith, G., and Stoker, C. (1949). Inhibition of crystalline lysozyme. Arch. Biochem. Biophys. 21: 383-94.
B-PER Bacterial Protein Extraction Reagents
I-PER Insect Cell Protein Extraction Reagent
Y-PER Yeast Protein Extraction Reagent
Inclusion Body Solubilization Reagent
Halt Protease Inhibitor Cocktail
Halt Phosphatase Inhibitor Cocktail