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Enhanced secretion of human α1-antitrypsin expressed with a novel glycosylation module in tobacco BY-2 cell culture.
- Source :
-
Bioengineered [Bioengineered] 2019 Dec; Vol. 10 (1), pp. 87-97. - Publication Year :
- 2019
-
Abstract
- Expression of recombinant proteins fused to a novel glycomodule tag, termed hydroxyproline (Hyp)-O-glycosylated peptides (HypGP), was earlier found to boost secreted protein yields up to 500-fold in plant cell culture. Here, this technology was applied to the expression of human protease inhibitor α1-antitrypsin (AAT) in tobacco BY-2 cell culture. A designer HypGP tag composed of a 'Ala-Pro' motif of 20 units, or (AP) <subscript>20</subscript> , was engineered either at the N- or C-terminal end of AAT. The (AP) <subscript>20</subscript> tag substantially increased the secreted yields of the recombinant AAT up to 34.7 mg/L. However, the (AP) <subscript>20</subscript> -tagged AAT products were frequently subjected to proteolytic processing. The intact AAT-(AP) <subscript>20</subscript> along with some of the truncated AAT domains exhibited desired biological activity in inhibiting elastase. The results from this research demonstrated that the designer (AP) <subscript>20</subscript> module engineered in BY-2 cells could function as a molecular carrier to substantially enhance the secreted yields of the recombinant AAT.
- Subjects :
- Base Sequence
Cell Culture Techniques
Dipeptides genetics
Dipeptides metabolism
Gene Expression
Glycosylation
Humans
Pancreatic Elastase metabolism
Peptides genetics
Peptides metabolism
Plant Cells metabolism
Plasmids chemistry
Plasmids metabolism
Proteinase Inhibitory Proteins, Secretory genetics
Proteinase Inhibitory Proteins, Secretory isolation & purification
Proteinase Inhibitory Proteins, Secretory pharmacology
Recombinant Fusion Proteins genetics
Recombinant Fusion Proteins isolation & purification
Recombinant Fusion Proteins pharmacology
Nicotiana cytology
Nicotiana metabolism
Transformation, Genetic
alpha 1-Antitrypsin genetics
alpha 1-Antitrypsin isolation & purification
alpha 1-Antitrypsin pharmacology
Pancreatic Elastase antagonists & inhibitors
Protein Processing, Post-Translational
Proteinase Inhibitory Proteins, Secretory biosynthesis
Recombinant Fusion Proteins biosynthesis
Nicotiana genetics
alpha 1-Antitrypsin biosynthesis
Subjects
Details
- Language :
- English
- ISSN :
- 2165-5987
- Volume :
- 10
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Bioengineered
- Publication Type :
- Academic Journal
- Accession number :
- 30957636
- Full Text :
- https://doi.org/10.1080/21655979.2019.1604037