Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10235462 | Process Biochemistry | 2014 | 6 Pages |
Abstract
Plant transient expression provides a rapid production platform for recombinant proteins but is linked with low protein yields. To test if plant-specific hydroxyproline (Hyp)-O-glycosylated peptide tags attached to a target protein can improve overall yields of recombinant protein transiently expressed in Nicotiana benthamiana, enhanced green fluorescence protein (EGFP) was expressed as a fusion with 5 or 32 tandem repeats of a serine-proline motif, designated (SP)5 or (SP)32, which is known to direct extensive Hyp-O-glycosylation in plants. EGFP containing the (SP)n motif showed enhanced yields in the order as follows: EGFP < EGFP-(SP)5 âªÂ (SP)5-EGFP < (SP)32-EGFP. The EGFP equivalent yield of (SP)32-EGFP was up to 16-fold greater than that of the EGFP control. In addition, both fully glycosylated (SP)32-EGFP (â¼115 kDa) and partially glycosylated (SP)32-EGFP (â¼40 kDa) were detected in protein extracts of N. benthamiana. These two types of glycoforms were completely segregated between media and cells in tobacco BY-2 cell cultures.
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Bioengineering
Authors
Maureen C. Dolan, Di Wu, Carole L. Cramer, Jianfeng Xu,