SAE0152

Lysozym aus Hühnereiweiss

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

• Synonym(e): Mucopeptid-N-Acetylmuramoylhydrolase, Muramidase
• CAS-Nummer: 12650-88-3
• EC-Nummer: 3.2.1.17 (BRENDA, IUBMB)
• MDL-Nummer: MFCD00131557
• UNSPSC-Code: 12352204
• NACRES: NA.77

Eigenschaften

• Biologische Quelle: chicken egg white
• Qualitätsniveau: 100
• Form: powder
• Spezifische Aktivität: ≥40000 units/mg protein
• Mol-Gew.: single-chain 14.3 kDa
• Leistungsmerkmale: DNA free
• Methode(n): cell based assay: suitable
• Eignung: suitable for cell lysis
• Anwendung(en): cell analysis
• Versandbedingung: wet ice
• Lagertemp.: −20°C

Beschreibung

Allgemeine Beschreibung

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.

Anwendung

Enzym bricht die Zellwände von Bakterien, wird verwendet um Sphäroplasten vorzubereiten.

• 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.

Biochem./physiol. Wirkung

Lysozym hydrolysiert β(1→4)-Bindungen zwischen N-Acetylmuraminsäure und N-Acetyl-D-Glucosamin-Resten in Peptidoglycan und zwischen N-Acetyl-D-Glucosamin-Resten in Chitodextrin. Gram-positive Zellen sind sehr anfällig für diese Hydrolyse, da ihre Zellwände einen hohen Peptidoglycan-Anteil haben.

Lysozym hydrolysiert β(1→4)-Bindungen zwischen N-Acetylmuraminsäure und N-Acetyl-D-Glucosamin-Resten in Peptidoglycan und zwischen N-Acetyl-D-Glucosamin-Resten in Chitodextrin. Gram-positive Zellen sind sehr anfällig für diese Hydrolyse, da ihre Zellwände einen hohen Peptidoglycan-Anteil haben. Gram-negative Bakterien sind auf Grund einer äußeren Membran und eines geringeren Peptidoglycan-Anteils weniger anfällig. Allerdings können diese Zellen in Gegenwart von EDTA, welches Chelate mit Metallionen in der äußeren bakteriellen Membran bildet, hydrolysiert werden.

Das Enzym ist über einen breiten pH-Bereich aktiv (6.0 bis 9.0). Bei pH 6.2 wird eine maximale Aktivität über einen größeren Ionenstärken-Bereich (0.02 bis 0.100 M), verglichen mit pH 9,2 (0.01 bis 0.06 M), beobachtet.

Leistungsmerkmale und Vorteile

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.

Einheitendefinition

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.

Sicherheitshinweise

• Piktogramme: GHS08
• Signalwort: Danger
• H-Sätze: H334
• P-Sätze: P261 - P284 - P501
• Gefahreneinstufungen: Resp. Sens. 1
• Lagerklassenschlüssel: 11 - Combustible Solids
• WGK: WGK 3
• Flammpunkt (°F): Not applicable
• Flammpunkt (°C): Not applicable