Isolation, characterization and evaluation of protein changes in the milk glands of honey bee at different ages using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)

Alpha-glucose oxidase, amylase and glucose oxidase, as part of digestive enzymes are the most important proteins in the milk glands of forager bees and expressed with three other proteins as the main protein for the production of royal jelly in worker bees. According to expression of proteins in different ages, effect on production of honey and royal jelly, the aim of this study was to investigate these changes. The proteins extracted from milk glands with four methods of tris, phosphate, tri-chloro acetic acid and phenol and stained with three methods included Cumasi Brilliant Blue R-250, Brilliant Blue G-250 and Silver nitrate. After determining the best method of extraction and staining, protein concentrations were be equal by Bradford method and their expressions were evaluated at five age period of birth, 5, 10, 15 and 20 days. The results showed that the highest protein expression involved in royal jelly of milk glands, corresponding to 10 days of age and the lowest protein expression was in 20 days. The highest expression level of proteins involved in honey production was in glucose oxidase, at 20 days and the lowest expression in amylase protein and glucose oxidase at the ages of birth, 5 and 10 days. Most of the major royal jelly protein expression was related to the ages of 5 and 10 days. The results show that there were large differences in the expression of these proteins in different ages that can affect the quality and quantity of honey and royal jelly.