SAE0152

Lysozyme from chicken egg white

free of DNA contaminants, suitable for Microbiome research, lyophilized powder, protein ≥90%, ≥40,000 units/mg protein

• Synonyme(s) : Mucopeptide N-acétylmuramoylhydrolase, Muramidase
• Numéro CAS: 12650-88-3
• Numéro de classification (Commission des enzymes): 3.2.1.17 (BRENDA, IUBMB)
• Numéro MDL: MFCD00131557
• Code UNSPSC : 12352204
• Nomenclature NACRES : NA.77

Propriétés

• Source biologique: chicken egg white
• Niveau de qualité: 100
• Forme: powder
• Activité spécifique: ≥40000 units/mg protein
• Poids mol.: single-chain 14.3 kDa
• Caractéristiques: DNA free
• Technique(s): cell based assay: suitable
• Adéquation: suitable for cell lysis
• Application(s): cell analysis
• Conditions d'expédition: wet ice
• Température de stockage: −20°C

Description

Description générale

Purified Lysozyme DNA free SAE0152 undergoes strict quality control testing to ensure the absence of detectable levels of contaminating DNA using 35 cycles PCR amplification of 16S and 18S rDNA using universal primer sets.

Lysozyme is a single chain polypeptide of 129 amino acids cross-linked with four disulfide bridges.1 It hydrolyzes b(1à 4) linkages between N-acetylmuraminic acid and N-acetyl-D-glucosamine residues in peptidoglycan and between N-acetyl-D-glucosamine residues in chitodextrin.2,3,4 The enzyme is often used for lysing bacterial cells by hydrolyzing the peptidoglycan present in the cell walls. Gram-positive cells are quite susceptible to this hydrolysis as their cell walls have a high proportion of peptidoglycan. Gram-negative bacteria are less susceptible due to the presence of an outer membrane and a lower proportion of peptidoglycan.
The enzyme is active over a broad pH range (6.0-9.0).

Lysozyme activity: ≥40,000 units/mg protein.

For isolation of nucleic acids, lysozyme has been used in the lysis peptidoglycan layer of bacterial cell walls.
The study of microbial communities has been revolutionized in recent years by the widespread adoption of culture independent analytical techniques such as 16S rRNA gene sequencing and metagenomics. Since DNA contamination during sample preparation is a major problem of these sequence-based approaches, DNA extraction reagents free of DNA contaminates are essential. This purified lysozyme preparation (SAE0152) is purified from chicken egg white, crystallized three times, dialyzed, and supplied as a lyophilized powder. Protein content by UV absorbance is ≥ 90% with the remainder (∼10%) being buffer salts such as sodium acetate and sodium chloride, and undergoes strict quality control testing to ensure the absence of detectable levels of contaminating DNA using 35 cycles PCR amplification of 16S and 18S rDNA using universal primer sets.

Application

• Lysozyme is used for the extraction of genomic DNA from bacterial cells.
• Lysozyme is used as an external standard for MALDI-TOF (matrix assisted laser desorption ionization-time of flight) mass analysis.
• Lysonzyme is used to prepare spheroplasts.

L′enzyme dégrade la paroi cellulaire des bactéries ; elle est utilisée pour préparer des sphéroplastes.

Actions biochimiques/physiologiques

Le lysozyme hydrolyse les liaisons β(1→4) entre les résidus d′acide N-acétylmuramique et de N-acétyl-D-glucosamine dans le peptidoglycane et entre les résidus de N-acétyl-D-glucosamine dans la chitodextrine. Les cellules à Gram positif sont très susceptibles à cette hydrolyse, car leur paroi cellulaire contient une proportion élevée de peptidoglycanes. Les cellules à Gram négatif sont moins susceptibles, en raison de la présence d′une membrane externe et d′une proportion plus faible de peptidoglycanes. Cependant, ces cellules peuvent être hydrolysées en présence d′EDTA, qui chélate les ions métalliques de la membrane externe des bactéries.

L′enzyme est active dans une vaste gamme de pH (de 6,0 à 9,0). À pH 6,2, une activité maximale est observée sur une plus vaste plage de forces ioniques (de 0,02 à 0,100 M) qu′à pH 9,2 (0,01 à 0,006 M).

Caractéristiques et avantages

Purified Lysozyme free SAE0152 undergoes strict quality control testing to ensure it will be Free of DNA contaminants, suitable for Microbiome research.

The study of microbial communities has been revolutionized in recent years by the widespread adoption of culture independent analytical techniques such as 16S rRNA gene sequencing and metagenomics. Since DNA contamination during sample preparation is a major problem of these sequence-based approaches, DNA extraction reagents free of DNA contaminates are essential.

Définition de l'unité

One unit will lyse 0.6 μg of Micrococcus lysodeikticus per minute by turbidimetric detection at 600 nm when suspended in buffer at pH 6.2 at 25 °C.

Informations sur la sécurité

• Pictogrammes: GHS08
• Mention d'avertissement: Danger
• Mentions de danger: H334
• Conseils de prudence: P261 - P284 - P501
• 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