Different plant species have been used as systems to produce recombinant proteins. Maize is a crop considered to have a large potential to produce high levels of recombinant proteins and is the host for the recombinant proteins from plants currently available on the market. In the development of a plant system to produce a recombinant proteins it is important to consider the costs related to downstream processing. Also, the steps necessary to achieve the protein purity required will be highly influenced by the quality of the extract obtained. In this study, we analyzed aqueous extracts from the endosperm of transgenic maize expressing recombinant human proinsulin (rhProinsulin). A study of the effects of the variables pH and ionic strength on the extraction efficiency was carried out using experimental design and response surface methodology. Besides the concentration of the recombinant protein, the characteristics of the extracts were evaluated in terms of concentration of native components (proteins, carbohydrates, and phenolic compounds) and extract filterability. The highest rhProinsulin concentration (97.33 ng/mL) was found with a 200 mM NaCl pH 10.0 extraction solution. Under this experimental condition the concentrations of total soluble proteins, carbohydrates, and phenolics were 2.01 mg/mL, 2.21 mg/mL, and 0.11 mmol/L, respectively.
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机译:不同的植物物种已被用作生产重组蛋白的系统。玉米是一种被认为具有产生高水平重组蛋白的潜力的作物,并且是来自市场目前可用的植物的重组蛋白的宿主。在生产重组蛋白的植物系统的发展中,重要的是考虑与下游加工有关的成本。而且,实现所需蛋白质纯度所需的步骤将受到所得提取物的质量的高度影响。在该研究中,我们分析了表达重组人胰岛素(RhProSillin)的转基因玉米胚乳的水性提取物。使用实验设计和响应表面方法进行了对变量pH和离子强度对提取效率的影响的研究。除了重组蛋白的浓度外,提取物的特征是在天然成分(蛋白质,碳水化合物和酚类化合物)的浓度方面进行评价,并提取过滤性。用200mM NaCl pH 10.0萃取溶液,发现最高的RhProInsulin浓度(97.33 ng / ml)。在该实验条件下,总可溶性蛋白质,碳水化合物和酚类的浓度分别为2.01mg / ml,2.21mg / ml和0.11mmol / L.
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