Lysozyme Human
Oryza sativa (rice).
EC 3.2.1.17, LYZ, Lysozyme.
Sterile Filtered lyophilized (freeze-dried) off white powder.
Purity as determined by SDS-PAGE is 85%.
THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals
Description
Recombinant Human Lysozyme produced in Plant is a non-glycosylated, polypeptide chain containing 130 amino acids and having a molecular mass of 14kDa. The Recombinant Human Lysozyme is purified by proprietary chromatographic techniques.
Product Specs
Lysozyme, an antimicrobial enzyme found in animals, forms a crucial part of the innate immune system. As a glycoside hydrolase, it breaks down the 1,4-beta-linkages between N-acetylmuramic acid and N-acetyl-D-glucosamine residues within peptidoglycan. This peptidoglycan is the primary component of gram-positive bacteria's cell walls. The main function of lysozymes is bacteriolytic, particularly those present in bodily tissues and fluids. They work in conjunction with the monocyte-macrophage system to enhance the effectiveness of immunoagents.
Produced from plants, Recombinant Human Lysozyme is a non-glycosylated polypeptide chain. It comprises 130 amino acids, resulting in a molecular mass of 14kDa. The purification of this recombinant enzyme is carried out using proprietary chromatographic techniques.
Sterile Filtered, lyophilized (freeze-dried) powder with a white appearance.
The protein (at a concentration of 1mg/ml) undergoes lyophilization without any additional additives.
To create stock solutions, gently dissolve the lyophilized powder in PBS (phosphate-buffered saline). Allow for several minutes of gentle mixing to ensure complete dissolution. The recommended stock concentration is 1mg/ml in PBS.
Lyophilized Lysozyme demonstrates stability at room temperature for up to 3 weeks. However, for extended storage, it's recommended to store it in a desiccated state below -18°C. After reconstitution, Recombinant Human Lysozyme should be stored at 4°C for a period of 2-7 days. For longer-term storage, keeping it below -18°C is advised. To further enhance its stability during long-term storage, consider adding a carrier protein such as HSA or BSA at a concentration of 0.1%. It's crucial to avoid repeated freeze-thaw cycles to maintain the enzyme's integrity.
The purity level, as determined by SDS-PAGE analysis, is 85%.
The biological activity is greater than 100,000 Units per mg of protein. One unit of activity is defined as the amount that produces a change in absorbance at 450nm (A450) of 0.001 per minute. This measurement is conducted at a pH of 6.24 and a temperature of 25°C, utilizing a suspension of Micrococcus lysodeikticus as the substrate in a reaction volume of 2.6ml.
Recombinant Human Lysozyme, at concentrations ranging from 0.05-0.5mg/ml (equivalent to 7,000-70,000 U/ml), is added to the cell extraction buffer for lysing bacterial cells. A common extraction buffer for E. coli comprises 0.2mg/ml of recombinant Lysozyme in a solution of 100mM Tris-HCl (pH 8), 2mM EDTA, and 0.05% Triton X-100. After suspending the cell paste in this extraction buffer, it undergoes incubation for a minimum of 15 minutes at room temperature. As an alternative, Triton X-100 at 1% and PMSF at 1mM can be introduced separately, followed by an incubation of the lysis solution for at least 15 minutes at 37°C. In certain instances, freeze-thaw cycles, sonication, or mechanical cell disruption methods are employed in conjunction with the lysozyme lysis protocol.
EC 3.2.1.17, LYZ, Lysozyme.
Oryza sativa (rice).
Product Science Overview
Lysozyme, also known as muramidase, is an enzyme that plays a crucial role in the innate immune system by breaking down the cell walls of bacteria. This enzyme was first discovered by Alexander Fleming in 1922 . Human lysozyme (hLYZ) is composed of 130 amino acids and contains four pairs of disulfide bonds, which contribute to its stability and antibacterial activity .
Lysozyme is a 1,4-β-N-acetylmuramidase that degrades the glycosidic bonds in the peptidoglycan of bacterial cell walls . This action is particularly effective against Gram-positive bacteria, making lysozyme a potent antibacterial agent. Human lysozyme is found in various bodily fluids, including tears, saliva, blood serum, and human milk .
Recombinant human lysozyme (rhLZM) is produced using genetic engineering techniques to express the human lysozyme gene in various host systems, such as bacteria, yeast, or mammalian cells. This recombinant form retains the same structure and function as the naturally occurring enzyme but can be produced in larger quantities and with greater purity .
Recombinant human lysozyme has shown significant potential in various therapeutic applications, particularly in wound management. Its ability to dissolve bacterial cell walls makes it an effective agent for treating wound infections and promoting wound healing . Current research focuses on developing carrier-based delivery systems, such as hydrogels, nanofilms, and electrospun fibrous membranes, to enhance the stability and controlled release of lysozyme at wound sites .
Despite its potential, the widespread clinical application of recombinant human lysozyme faces several challenges. These include ensuring the stability of the enzyme during storage and delivery, as well as overcoming any potential immunogenicity issues. Ongoing research aims to address these challenges by developing advanced delivery systems and exploring new therapeutic applications .
