کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
6488114 1416357 2018 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Effects of N-glycosylation on the biochemical properties of recombinant bEKL expressed in Pichia pastoris
موضوعات مرتبط
مهندسی و علوم پایه مهندسی شیمی بیو مهندسی (مهندسی زیستی)
پیش نمایش صفحه اول مقاله
Effects of N-glycosylation on the biochemical properties of recombinant bEKL expressed in Pichia pastoris
چکیده انگلیسی
Enterokinase is an ideal tool protease for cleaving fusion proteins in genetic engineering. The bovine enterokinase light chain (bEKL) produced in Pichia pastoris was shown to be a glycoprotein. To study the effects of N-glycosylation on the biochemical properties of bEKL, the enzyme was deglycosylated via site-directed mutagenesis. The results showed that elimination of the N-glycosylation sites of bEKL (N64, N103 and N165) did not significantly affect the protein secretion level in P. pastoris, but it does greatly influence its enzymatic activity. The N64Q increased the specific activity of the enzyme for GD4K-β-naphthylamide and improved its catalytic efficiency. Moreover, the glycosylated bEKL is more thermostable than its deglycosylated counterparts. Structural analysis of glycosylated and deglycosylated bEKL revealed that the removal of N-glycosylation did not have pronounced changes on the secondary structure but there was a significant difference in the tertiary structure. In conclusion, this study demonstrated that the effects of glycosylation at different degrees and sites in bEKL were diverse. Moreover, this work will provide theoretical support for designing enzymes on the basis of N-glycosylation to meet the demands of the biochemical industry.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Enzyme and Microbial Technology - Volume 114, July 2018, Pages 40-47
نویسندگان
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