Effect of nitrate concentration in nutrient solution on hemagglutinin content of Nicotiana benthamiana leaves in a viral vector-mediated transient gene expression system
Effect of nitrate concentration in nutrient solution on hemagglutinin content of Nicotiana benthamiana leaves in a viral vector-mediated transient gene expression system
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DOI：
10.5511/plantbiotechnology.14.0403a
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发表时间：
2014-01-01
影响因子：
1.6
通讯作者：
Fujiwara, Kazuhiro
中科院分区：
文献类型：
--
作者：
Fujiuchi, Naomichi;Matsuda, Ryo;Fujiwara, Kazuhiro
病毒载体介导的瞬时表达使利用整个植物快速生产生物药物蛋白成为可能。近年来，植物生长条件作为影响重组蛋白含量的因素受到了人们的密切关注。特别是，由于氮素是蛋白质合成的重要来源，供应给根区的氮素可能会影响重组蛋白质的积累。我们测试了在农杆菌介导的烟草病毒载体(农业渗入)之前，在植物生长期间用高硝酸盐营养液施肥是否导致单位叶鲜重和/或每株的高重组流感血凝素(HA)含量。与含12或36 mM硝酸盐的营养液相比，含60 mM硝酸盐的营养液单位叶鲜重的HA含量高40%，单株HA含量相当。腐殖酸、总可溶性蛋白(TSP)和可溶性还原氮含量之间呈正相关，说明高硝酸盐营养液施肥增加了可溶性还原氮，从而提供了较高的TSP和HA。尽管单位叶片鲜重增加的HA含量被每株叶片鲜重的减少所抵消，但富硝酸盐施肥是一种容易应用的技术，可以从较小的叶片生物量中获得给定数量的HA，从而潜在地降低使用病毒载体介导的瞬时表达系统生产HA疫苗的下游加工成本。
Viral vector-mediated transient expression enables rapid biopharmaceutical protein production using whole plants. In recent years, plant growth conditions have received close attention as factors influencing the content of recombinant protein. In particular, because nitrogen is an important resource used in protein synthesis, nitrogen supplied to the root zone may influence accumulation of recombinant proteins. We tested whether fertilization of Nicotiana benthamiana with high-nitrate nutrient solution during plant growth prior to Agrobacterium-mediated delivery (agroinfiltration) of a tobamoviral vector resulted in high recombinant influenza hemagglutinin (HA) content per unit of leaf fresh weight and/or per plant. Nutrient solution containing 60 mM nitrate yielded 40% higher HA content per unit of leaf fresh weight, and comparable HA content per plant, relative to nutrient solution containing 12 or 36 mM nitrate. There were positive correlations among HA, total soluble protein (TSP), and soluble reduced-nitrogen content, suggesting that fertilization with the high-nitrate nutrient solution increased soluble reduced nitrogen and subsequently provided high TSP and HA. Although the increased HA content per unit of leaf fresh weight was offset by reduced leaf fresh weight on a per plant basis, nitrate-enriched fertilization represents an easily applicable technique to obtain a given amount of HA from smaller leaf biomass, thereby potentially reducing downstream processing costs for HA vaccine production using viral vector-mediated transient expression systems.