| Popis: |
Insects are protein-rich and some edible species are used worldwide as food and feed. Therefore, insect proteins are of great interest for cellular agriculture (CA), i.e. tissue engineering for food production, particularly for cultivated meat production. Because of their significant role in amino acid metabolism, storage proteins stand out. However, they have not been sufficiently investigated, hence it is crucial to characterize them. The first step is the successful isolation of individual proteins of interest. In our previous work, the optimization of the method isolating individual hemolymph proteins from native polyacrylamide gel (nPAG) was shown using diapausing 5th instar larvae of the economically important Ostrinia nubilalis as a model system. The main objective in this study was the validation of the described method by isolating specific proteins of interest from Tenebrio molitor and Zophobas morio larvae hemolymph, as well as from the whole-body samples of Plodia interpunctella larvae. Similarly to O. nubilalis, P. interpunctella is an economically important pest species, while T. molitor and Z. morio larvae have been declared edible. Total protein concentrations were determined by Bradford protein assay and the results showed that the samples of Z. morio are the richest in total proteins, followed by T. molitor and P. interpunctella, respectively. Proteins were separated by discontinuous native polyacrylamide gel electrophoresis (nPAGE) and protein fractions of interest were cut from the nPAG and eluted overnight from the gels. To confirm that the proteins were well isolated, the individual fractions were run on discontinuous nPAGE. In total, six protein fractions were successfully isolated – two from every analysed insect species, respectively, and in sufficient amounts for downstream applications in CA. |
