Production of biologically active human thioredoxin 1 protein in lettuce chloroplasts
Production of biologically active human thioredoxin 1 protein in lettuce chloroplasts
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DOI：
10.1007/s11103-011-9745-5
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发表时间：
2011-07-01
影响因子：
5.1
通讯作者：
Yokota, Akiho
中科院分区：
文献类型：
--
作者：
Lim, Soon;Ashida, Hiroki;Yokota, Akiho
在植物中生产人类治疗性蛋白质为低成本生产提供了机会，并最大限度地减少了潜在人类病原体污染的风险。叶绿体基因工程是一个特别有前途的策略，因为植物叶绿体可以产生大量的外源目的蛋白。氧化应激是多种人类疾病的关键因素。人硫氧还蛋白1（hTrx 1）是一种应激诱导蛋白，其作为抗氧化应激的抗氧化剂起作用，并且hTrx 1的过表达已被证明可以抑制小鼠中的多种疾病。因此，hTrx 1是一种有前景的候选人作为一种新的人类治疗蛋白。我们创建了在psbA启动子控制下表达hTrx 1的转质体莴苣。转质体植株生长正常，可育。在成熟叶片中，hTrx 1蛋白的积累量约占可溶性蛋白总量的1%。从莴苣叶中纯化的hTrx 1蛋白具有功能活性，并减少二硫化物胰岛素。纯化的蛋白质保护小鼠胰岛素瘤细胞系6细胞免受过氧化氢的损伤，如先前报道的大肠杆菌中表达的重组hTrx 1。这是第一个报告的生物活性hTrx 1蛋白在植物叶绿体中的表达。这项研究为基于植物的hTrx 1生产开辟了可能性。考虑到该表达宿主是可食用作物，该转质体莴苣可能适合于口服递送hTrx 1。
The production of human therapeutic proteins in plants provides opportunities for low-cost production, and minimizes the risk of contamination from potential human pathogens. Chloroplast genetic engineering is a particularly promising strategy, because plant chloroplasts can produce large amounts of foreign target proteins. Oxidative stress is a key factor in various human diseases. Human thioredoxin 1 (hTrx1) is a stress-induced protein that functions as an antioxidant against oxidative stress, and overexpression of hTrx1 has been shown to suppress various diseases in mice. Therefore, hTrx1 is a prospective candidate as a new human therapeutic protein. We created transplastomic lettuce expressing hTrx1 under the control of the psbA promoter. Transplastomic plants grew normally and were fertile. The hTrx1 protein accumulated to approximately 1% of total soluble protein in mature leaves. The hTrx1 protein purified from lettuce leaves was functionally active, and reduced insulin disulfides. The purified protein protected mouse insulinoma line 6 cells from damage by hydrogen peroxide, as reported previously for a recombinant hTrx1 expressed in Escherichia coli. This is the first report of expression of the biologically active hTrx1 protein in plant chloroplasts. This research opens up possibilities for plant-based production of hTrx1. Considering that this expression host is an edible crop plant, this transplastomic lettuce may be suitable for oral delivery of hTrx1.