Экспрессия и очистка бактериальной глюкозодегидрогеназы и гибридного белка GDH-ab в дрожжах Sacharomyces Cerevisi AE, выделение гомогенных белковых препаратов

Abstract

Eukaryotic expression system, bearing bacterial gene of glucose dehydrogenase alone or fused with amyloid peptide coding sequence, were constructed by the using gene engineering methods. Investigation of heterologous gene expression in yeast S. cerevisiae allowed to optimize cultivation of producents in order to achieve maximal amount of active protein. The maximal GDH activity in the membraneous fraction of secretion systems possesing transformants was detected. The level of the expressed in S. cerevisiae and purified on CM column GDH and GDH-ab protein reached 15-20mg from 1 g wet weight of yeast culture.