Abstract
Plants are an attractive host system for pharmaceutical protein production. Many therapeutic proteins have been produced and scaled up in plants at a low cost compared to the conventional microbial and animal-based systems. The main technical challenge during this process is to produce sufficient levels of recombinant proteins in plants. Low yield is generally caused by proteolytic degradation during expression and downstream processing of recombinant proteins. The yield of human therapeutic interleukin (IL)-10 produced in transgenic tobacco leaves was found to be below the critical level, and may be due to degradation by tobacco proteases. Here, we identified a total of 60 putative cysteine protease genes (CysP) in tobacco. Based on their predicted expression in leaf tissue, 10 candidate CysPs (CysP1-CysP10) were selected for further characterization. The effect of CysP gene silencing on IL-10 accumulation was examined in tobacco. It was found that the recombinant protein yield in tobacco could be increased by silencing CysP6. Transient expression of CysP6 silencing construct also showed an increase in IL-10 accumulation in comparison to the control. Moreover, CysP6 localizes to the endoplasmic reticulum (ER), suggesting that ER may be the site of IL-10 degradation. Overall results suggest that CysP6 is important in determining the yield of recombinant IL-10 in tobacco leaves.
Highlights
Plants are attractive biofactories for recombinant protein production
A total of 55 putative cysteine protease genes (CysP) genes were identified in the database using the key word ‘cysteine protease’
Out of 55 putative CysP sequences, 32 were tentative contig sequences which were created by assembling expressed sequence tags (ESTs) into virtual transcripts
Summary
Plants are attractive biofactories for recombinant protein production. They offer several advantages such as reduction in cost of production and increased scalability over microbial and mammalian cell culture systems. This low cost of large scale protein production is due to significant reduction in capital investment and processing cost. Unlike cell culture based systems, plants do not require fermenters or highly skilled personnel to operate them [1]. Plants are capable of carrying out complex post translational modifications (PTM), a process necessary for the biological activity of many eukaryotic proteins.
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