Protein characterization. The lytic activities of lysozymes were analyzed by light scattering assays as previously decribed .
Commercially sourced human lysozyme .. and hen egg white ly….. Were use as controls.
Kinetic assays used ….. luteus.. enzymes …….. plates were assessed .. decrease in absorbance at 450 ..30.. Spectra Max 190 plate reader at….. specific rates were calculated from the initial linear protion of the curve . Pseudo Michaelis-Menten kinetics were assessed at four differrnt substrate concentrations
With 14 nM enzymes … parametars … nonlinear regression using version (GraphPad Software inc ).. ..
The presence of Ivyc,
, where .. initial rate in the presence of inhibitor, .. absence of inhibitor, .. total inhibitor
Melting t… were measured using differrntial scanning fluorimetry. Lysomes and SYPRO Orange (Invitrogen) prepared in …..
Samples were run on … and analyzed using Prism and Excel following the protocols provided by Niesen ey=t al
Quantitative Culture. Single colonies of E.coli … inoculated in 5 mL of LB and grow …. Overnight. Overnights were subcutured 1:100 into …mid log . Subsequently, cells were dilted either control .. mediluted of working solution
.. respectively. Cell suspensions were incubated ,.., serially diluted, and colonies were enumerated the next morning.
The bactericidal activities of lysozymes were reprwsented by percentage of killing,