Departamento de Processos Biotecnológicos, Faculdade de Engenharia Química, Universidade Estadual de Campinas, CP 6066, CEP 13083-970, Campinas, SP, Brazil.
Biotechnol Prog. 2011 Jul;27(4):1029-35. doi: 10.1002/btpr.627. Epub 2011 May 27.
The use of recombinant proteins has increased greatly in recent years, as also have increased the number of techniques and materials used for their production and purification. Among the different types of bioreactors being studied, there is a general consensus among scientists that production in green plant tissues such as leaves is more feasible. However, the presence of chlorophyll and phenolic compounds in plant extracts, which can precipitate and denature the proteins besides damaging separation membranes and gels, makes this technology impracticable on a commercial scale. In the present work, the adsorption to electrochemically produced aluminum hydroxide gel was applied as a prepurification step for recombinant synthetic green fluorescent protein (sGFP), also referred to as enhanced green fluorescent protein, produced in Nicotiana benthamiana leaves. Removal efficiencies of 99.7% of chlorophyll, 88.5% of phenolic compounds, and 38.5% of native proteins from the N. benthamiana extracts were achieved without removing sGFP from the extracts. As electrochemical preparation of aluminum hydroxide gel is a cost-effective technique, its use can substantially contribute to the development of future production platforms for recombinant proteins produced in green plant tissues of pharmaceutical and industrial interest.
近年来，由于用于生产和纯化的技术和材料数量增加，重组蛋白的使用大大增加。在研究的不同类型的生物反应器中，科学家普遍认为在绿色植物组织（如叶片）中生产更可行。然而，植物提取物中叶绿素和酚类化合物的存在会沉淀和变性蛋白质，除了损坏分离膜和凝胶外，这使得该技术在商业规模上不可行。在本工作中，吸附到电化学产生的氢氧化铝凝胶上被用作从 Nicotiana benthamiana 叶片中生产的重组合成绿色荧光蛋白（sGFP）的预纯化步骤，也称为增强型绿色荧光蛋白。从提取物中去除了 99.7%的叶绿素、88.5%的酚类化合物和 38.5%的天然蛋白质，而不会从提取物中去除 sGFP。由于电化学制备氢氧化铝凝胶是一种具有成本效益的技术，因此其使用可以为未来在具有医药和工业价值的绿色植物组织中生产重组蛋白的生产平台的发展做出重大贡献。
