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首页> 外文期刊>Biotechnology Progress >Aqueous Extraction of Recombinant Human Proinsulin from Transgenic Maize Endosperm
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Aqueous Extraction of Recombinant Human Proinsulin from Transgenic Maize Endosperm

机译：从转基因玉米胚乳中提取重组人胰岛素原的水溶液

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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.
机译：不同的植物种类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潜力的作物，并且是目前市场上出售的植物中重组蛋白的宿主。生产重组蛋白的植物系统，重要的是要考虑与下游加工有关的成本。此外，获得所需蛋白纯度所需的步骤将受到所得提取物质量的极大影响。表达重组人胰岛素原（rhProinsulin）的转基因玉米胚乳中的提取物。使用实验设计和响应面方法研究了pH和离子强度变量对提取效率的影响。除重组蛋白的浓度外，提取物的特征以天然成分的浓度（p蛋白质，碳水化合物和酚类化合物）和提取物的过滤性能。在200 mM NaCl pH 10.0提取溶液中发现最高的rhproinsulin浓度（97.33 ng / mL）。在此实验条件下，总可溶性蛋白，碳水化合物和酚类的浓度为分别为2.01 mg / mL，2.21 mg / mL和0.11 mmol / L。

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