Robust expression of a bioactive mammalian protein in Chlamydomonas chloroplast
Robust expression of a bioactive mammalian protein in Chlamydomonas chloroplast
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DOI：
10.1111/j.1467-7652.2007.00249.x
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发表时间：
2007-05-01
影响因子：
13.8
通讯作者：
Mayfield, Stephen P.
中科院分区：
文献类型：
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作者：
Manuell, Andrea L.;Beligni, Maria Veronica;Mayfield, Stephen P.
我们已经使真核藻的叶绿体产生了许多重组蛋白，包括人类的单抗，但到目前为止，只实现了总蛋白的0.5%的表达。在这里，我们表明，通过工程牛乳房相关血清淀粉样蛋白(M-SAA)的哺乳动物编码区作为叶绿体PSBA编码区的直接替代，我们可以获得占总蛋白5%以上的重组蛋白的表达。叶绿体表达的M-SAA主要以可溶性蛋白质的形式积累，含有正确的氨基末端序列，很少或没有翻译后修饰。M-SAA存在于哺乳动物的初乳中，可刺激肠道粘蛋白的产生，预防细菌和病毒感染。叶绿体表达和纯化的M-SAA能够刺激人肠道上皮细胞系产生粘蛋白。由于莱茵衣藻是一种可食用的藻类，在这种生物中生产治疗蛋白为口服肠道活性蛋白提供了可能性，如M-SAA。
We have engineered the chloroplast of eukaryotic algae to produce a number of recombinant proteins, including human monoclonal antibodies, but, to date, have achieved expression to only 0.5% of total protein. Here, we show that, by engineering the mammalian coding region of bovine mammary-associated serum amyloid (M-SAA) as a direct replacement for the chloroplast psbA coding region, we can achieve expression of recombinant protein above 5% of total protein. Chloroplast-expressed M-SAA accumulates predominantly as a soluble protein, contains the correct amino terminal sequence and has little or no post-translational modification. M-SAA is found in mammalian colostrum and stimulates the production of mucin in the gut, acting in the prophylaxis of bacterial and viral infections. Chloroplast-expressed and purified M-SAA is able to stimulate mucin production in human gut epithelial cell lines. As Chlamydomonas reinhardtii is an edible alga, production of therapeutic proteins in this organism offers the potential for oral delivery of gut-active proteins, such as M-SAA.