Transfer of egg white proteins with reference to lysozyme during the development of Meleagris gallopavo (Galliformes: Phasianidae) embryos

The molecular weight is 13.6 KDa and the electrophoretic band that appears to move from egg whites to other egg chambers is considered to be a lysozyme band. To confirm the appearance and metastasis of lysozyme, lysozyme activity was measured using Micrococcus lysodeikticus as a substrate. In egg white, lysozyme activity was found to be highest in all egg chambers (Fig. 2A-E). It begins on day 0 and lasts until day 26 (FIG. 2A; supplementary table I). Maximum enzyme activity was seen at day 24 in all developmental stages and then declined.

In the extraembryonic fluid, lysozyme activity was detected on days 15-18 and increased as the development progressed (FIG. 2B; supplementary Table II). For amniotic fluid, enzyme activity was measured on day 15 and continued on day 24, with maximum enzyme activity, until the last day of liquid emergence (FIG. 2C; supplementary table III). The putative electrophoretic lysozyme band appeared at day 15 (FIG. 1D), but the activity of lysozyme in intestinal fluid was first detected on day 16 (FIG. 2D). The appearance of lysozyme activity and the appearance of lysozyme band can be due to very low concentrations of enzyme that can not be detected by lysozyme activity assay. The activity of enzymes in intestinal juice fluctuated during development; however, it was generally high at the end of the incubation period and reached a maximum on the last hatching day (Day 28; Figure 2D; Supplementary Table IV). After rupture and rupture of Cloaka, the lysozyme activity in the residual fluid remaining in the intestine decreases

In thin egg yolks, the putative lysozyme band was hardly visible during development (FIG. 1E) and lysozyme activity was not detected throughout the liquid appearance. In thick egg yolks, a weakly estimated lysozyme band appeared early in development (FIG. 1F), lysozyme activity was measured on day 17 and continued thereafter (FIG. 2E; supplementary table V). In general, the activity of lysozyme increases as development progresses, reaching its maximum value after hatching. Changes in enzyme activity in all egg chambers indicated significant differences in enzyme activity between different sample groups at different developmental stages (FIGS. 2A-E).

Lysozyme used for quantification of egg white (HEW), a valuable protein in the food industry and pharmaceutical industry, has also been developed by indirect enzyme immunoassay (ELISA) which is also used in poultry research. Various experimental conditions (coating, antibody dilution, sample dilution, preparation, blocking agent and incubation time) were determined to optimize the assay to quantify HEW in egg white samples. HEW samples were diluted 1: 3000 to avoid possible matrix effects that could be caused by the interaction of lysozyme with other egg white proteins. The linear range of lysozyme was 0.38 to 4.8 μg / mL, the internal and experimental variability was 6.8% and 7.6%, respectively, and the lower limit of detection was 0.264 μg / mL. The concentration of lysozyme in the egg production protein is in the range of 2.2 to 4.5 mg / mL, emphasizing mutual variability.

In this experiment, lysozyme was purified from egg white using CM Sepharose based ion exchange chromatography. The protein consists of a complex mixture of proteins such as ovalbumin, lysozyme, albumin, ovotransferrin, flavoprotein, G 2 globulin and G 3 globulin. Among them, lysozyme is a low molecular weight protein (molecular weight 14.3 kDa), which is a commercially important enzyme used as an effective antimicrobial agent, food additive and antiulcer agent and precursor of infectious drug is there. (Yilmaz, Bayramoglu, and Arca 2005). This experiment was designed to demonstrate the practical application of CM Sepahalose resin in the isolation of lysozyme from whole albumen. Here, lysozyme is chosen as the basic protein and the pH is maintained at 2 with Tris buffer and sodium chloride.

This laboratory study examined purification and analysis of egg white lysozyme. Lysozyme is both a protein and an enzyme that catalyze 1,4-β-linked hydrolysis between N-acetylmuramic acid and N-acetyl-D-glucosamine residues in peptidoglycan (Lodish et al. , 2000). It abundantly exists in protein (egg white), which protects egg embryos from invasion of bacteria. Several measurements were made in this laboratory experiment and each assay showed significant properties of lysozyme.