Abstract

Agrobacterium-mediated transient expression systems enable plants to produce a wide range of recombinant proteins on a rapid timescale. To achieve economically feasible upstream production and downstream processing, two yield parameters should be considered: (1) recombinant protein content per unit biomass and (2) recombinant protein productivity per unit area–time at the end of the upstream production. Because environmental factors in the upstream production have impacts on these parameters, environment control is important to maximize the recombinant protein yield. In this review, we summarize the effects of pre- and postinoculation environmental factors in the upstream production on the yield parameters and discuss the basic concept of environment control for plant-based transient expression systems. Preinoculation environmental factors associated with planting density, light quality, and nutrient supply affect plant characteristics, such as biomass and morphology, which in turn affect recombinant protein content and productivity. Accordingly, environment control for such plant characteristics has significant implications to achieve a high yield. On the other hand, postinoculation environmental factors, such as temperature, light intensity, and humidity, have been shown to affect recombinant protein content. Considering that recombinant protein production in Agrobacterium-mediated transient expression systems is a result of a series of complex biological events starting from T-DNA transfer from Agrobacterium tumefaciens to protein biosynthesis and accumulation in leaf tissue, we propose that dynamic environment control during the postinoculation process, i.e., changing environmental conditions at an appropriate timing for each event, may be a promising approach to obtain a high yield. Detailed descriptions of plant growth conditions and careful examination of environmental effects will significantly contribute to our knowledge to stably obtain high recombinant protein content and productivity, thus enhancing the utility of plant-based transient expression systems as recombinant protein factories.

Highlights

  • Plants offer several benefits for recombinant protein production, including the potential for low-cost and large-scale biomass production, the low risk of contamination with human pathogens or toxins, and the ability to perform posttranslational protein modifications (Fischer et al, 2013)

  • We focus on the effects of environmental factors during the upstream production on recombinant protein content and productivity in Agrobacterium-mediated transient expression systems

  • Cazzonelli and Velten (2006) observed that the recombinant luciferase content in N. tabacum leaves treated in the dark was 20% of that treated at a photosynthetic photon flux density (PFD) (PPFD) of 80–100 μmol m−2 s−1

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Summary

INTRODUCTION

Plants offer several benefits for recombinant protein production, including the potential for low-cost and large-scale biomass production, the low risk of contamination with human pathogens or toxins, and the ability to perform posttranslational protein modifications (Fischer et al, 2013). Pharmaceutical-grade plant-based recombinant proteins can be produced at a commercial scale in closed facilities enabling good manufacturing practice-compliant processes (Fischer et al, 2012; Warzecha, 2012) Given these benefits, transient expression systems have opened a new avenue for the production of niche biopharmaceuticals and low-cost enzymes, such as individualized vaccines (McCormick et al, 2008), emerging disease vaccines (Shoji et al, 2009; D’Aoust et al, 2010), industrial enzymes (Hwang et al, 2012), and research reagents (Pogue et al, 2010). We discuss postinoculation environmental effects on recombinant protein content and productivity from biological aspects involved in protein biosynthesis and metabolism

EFFECTS OF PREINOCULATION ENVIRONMENTAL FACTORS
EFFECTS OF POSTINOCULATION ENVIRONMENTAL FACTORS
Light Intensity
Light intensity
CONCLUSION AND FUTURE DIRECTIONS
Findings
AUTHOR CONTRIBUTIONS
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