원문정보
초록
영어
The glucocerebrosidase (GCR) is a lysosomal enzyme which catalyzes glycolipid glucocerebroside to glucose and ceramide. This enzyme is a glycoprotein glycosylated by oligosaccharide chain including N-acetylglucosamine, galactose, fucose and mannose. It is well-known that the deficiency of GCR causes Gaucher disease which showed hepatomegaly, bone lesion, pulmonary disease and neurological disease. To treat Gaucher disease, ERT (Enzyme Replacement Therapy) has been used to supplement GCR. In this study, we attempt to express the GCR in Pichia pastoris, methylotrophic yeast. Since P. pastoris has the characteristics of both eukaryote and prokaryote, GCR retained biologically same activity can be produced with high titer. Also, we have carried out an experiment on the expression of GCR having different signal peptide and using Kozak consensus sequence to enhance expression level. Two products, intracellular type and secretion type of GCR obtained from flask culture using methanol induction were analyzed by SDS-PAGE and Western blot.