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• W2901935707 startingPage "1130" @default.
• W2901935707 abstract "Summary The key technical bottleneck for exploiting plant hairy root cultures as a robust bioproduction platform for therapeutic proteins has been low protein productivity, particularly low secreted protein yields. To address this, we engineered novel hydroxyproline (Hyp)‐ O ‐glycosylated peptides (Hyp GP s) into tobacco hairy roots to boost the extracellular secretion of fused proteins and to elucidate Hyp‐ O ‐glycosylation process of plant cell wall Hyp‐rich glycoproteins. Hyp GP s representing two major types of cell wall glycoproteins were examined: an extensin module consisting of 18 tandem repeats of ‘Ser‐Hyp‐Hyp‐Hyp‐Hyp’ motif or ( SP 4) 18 and an arabinogalactan protein module consisting of 32 tandem repeats of ‘Ser‐Hyp’ motif or ( SP ) 32 . Each module was expressed in tobacco hairy roots as a fusion to the enhanced green fluorescence protein ( EGFP ). Hairy root cultures engineered with a Hyp GP module secreted up to 56‐fold greater levels of EGFP , compared with an EGFP control lacking any Hyp GP module, supporting the function of Hyp GP modules as a molecular carrier in promoting efficient transport of fused proteins into the culture media. The engineered ( SP 4) 18 and ( SP ) 32 modules underwent Hyp‐ O ‐glycosylation with arabino‐oligosaccharides and arabinogalactan polysaccharides, respectively, which were essential in facilitating secretion of the fused EGFP protein. Distinct non‐Hyp‐ O ‐glycosylated ( SP 4) 18 ‐ EGFP and ( SP ) 32 ‐ EGFP intermediates were consistently accumulated within the root tissues, indicating a rate‐limiting trafficking and/or glycosylation of the engineered Hyp GP modules. An updated model depicting the intracellular trafficking, Hyp‐ O ‐glycosylation and extracellular secretion of extensin‐styled ( SP 4) 18 module and AGP ‐styled ( SP ) 32 module is proposed." @default.
• W2901935707 created "2018-11-29" @default.
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• W2901935707 date "2018-12-14" @default.
• W2901935707 modified "2023-10-13" @default.
• W2901935707 title "Engineering ‘designer’ glycomodules for boosting recombinant protein secretion in tobacco hairy root culture and studying hydroxyproline‐ <i>O</i> ‐glycosylation process in plants" @default.
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• W2901935707 doi "https://doi.org/10.1111/pbi.13043" @default.
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