canine milk lysozyme exhibits charge heterogeneity after sample purification. Four Asn residues deamidate rapidly under mild conditions of pH 8.0 and 30°C. One Asn residue, which is stable in the native state, is labile to deamidation in the unfolded state
hydrophobic affinity ligand L-tryptophan immobilized magnetic poly(glycidyl methacrylate) [m-poly(GMA)] beads in monosize form (0.0016 mM in diameter) are used for the affinity purification of lysozyme from chicken egg white. The chemisorption processes can be the rate-limiting step in the adsorption process. After 10 adsorption-elution cycles, m-poly(GMA)-L-tryptophan beads can be used without significant loss in lysozyme adsorption capacity
native isozymes to homogeneity by ammonium sulfate fractionation, two steps of cation exchange chromatography, and isozyme separation by reversed phase gel filtration
purification of recombinant mutants in unfolded non-native state, solubilization of the recombinant mutants from Escherichia coli strain BL21(DE3) inclusion bodies in 20 mM Tris-HCl, 50 mM NaCl, 5 mM EDTA, and 8 M urea, pH 7.5, centrifugation, and cation exchange chromatography with elution by 50 mM Tris-HCl, 50 mM NaCl, 1 mM EDTA, and 4 M urea, pH 7.5
recombinant enzyme from sugarcane stalks, Saccharum spp. hybrid, extraction condition evaluation and optimization at 22°C, and different pH levels and salt concentrations, overview. pH values greater than 7.5 and salt concentrations higher than 150 mM NaCl do not increase the amount of extracted enzyme. Purification by cross-flow filtration and hydrophobic interaction chromatography.
recombinant enzyme from transgenic cow milk, from which fat and casein has been removed for purification of the enzyme, by cation exchange chromatography and gel filtration to over 95% purity
recombinant enzyme fron Escherichia coli, molecular interactions between lysozyme and a His-tagged inhibitor Ivy allows for one-step purification by immobilized nickel affinity chromatography
recombinant GST-tagged enzyme from Escherichia coli inclusion bodies by solubilization and refolding, GST-tag cleavage through thrombin, and anion exchange chromatography and RPLC gel filtration
recombinant wild-type and mutant MdL2s from Pichia pastoris by ammonium sulfate fractionation, and cation exchange, for the sextuple mutant, or anion exchange, for the other mutants, chromatography
the cell pellet is resuspended in 50 ml lysis buffer (20 mM Tris-HCl pH 8.0 and 300 mM sodium chloride) and lysed by sonication on ice. All subsequent purification steps are performed at 277 K. The lysate is centrifuged at 15 000g for 30 min. The supernatant is applied onto a 10 ml Ni2+-NTA affinity column (Qiagen) equilibrated with lysis buffer. Nonspecifically bound proteins are washed from the column using 200 ml lysis buffer containing 15 mM imidazole. The recombinant protein is then eluted from the column with 20 ml elution buffer containing 20 mM Tris-HCl pH 8.0, 300 mM sodium chloride and 400 mM imidazole. The protein is concentrated and buffer-exchanged to the final buffer (5 mM Tris-HCl pH 8.0 and 10 mM sodium chloride). The final purified protein concentration was about 50 mg/ml and the purity is determined by SDS-PAGE to be about 95%
the insoluble protein is solubilized in urea and purified by passing through a His-bind column, and the lytic activity is then restored by a refolding process
two serine-rich heptapeptides, Ser-Ser-Ser-Lys-Ser-Ser-Ser (S6K) and Ser-Ser-Ser-Ser-Ser-Ser-Ser (S7) are fused to the C-terminus of chicken lysozyme by genetic modification. The cDNAs of S6K-lysozyme and S7-lysozyme are inserted into the expression vector of Pichia pastoris and secreted in the yeast cultivation medium
3.2.1.17 lysozyme
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