Recombinant polypeptide production in E. coli: towards a rational approach to improve the yields of functional proteins.
Recombinant polypeptide production in E. coli: towards a rational approach to improve the yields of functional proteins.
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DOI：
10.1186/1475-2859-12-101
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发表时间：
2013-11-01
影响因子：
6.4
通讯作者：
de Marco A
中科院分区：
文献类型：
--
作者：
de Marco A
互补技术的发展使得在细菌中成功地生产重组多肽，并为生物学研究人员开辟了新的途径，使他们的实验工作所需的适当数量的蛋白质变得容易、快速和廉价。然而，重组方法在某种程度上仍然是不可预测的，因为许多结构抗拒明显的生产或作为集合积累。确定了几种促进天然样蛋白积累的因素和物理/化学条件。与此同时，人们也承认，大多数折叠的结果是不稳定的，几乎任何蛋白质都需要自己特定的优化条件来实现正确的折叠。了解重组蛋白生产的特定关键点的尝试错过了设定普遍有用的协议的目标，但通过提出新的经验组合，有助于提高成功率。尽管如此，最近发表在文献中的结果使我们能够更好地理解大肠杆菌中控制蛋白质生产的一些关键机制，并能够阐明合理的方法，以提高所生产多肽的定量和定性特征。这一结果将在确定妨碍任何单个构造完成本机折叠的限制步骤变得简单时实现。本文将讨论表达率、折叠机制和分泌效率等因素对最终蛋白产量的影响。
The development of complementary technologies enabled the successful production of recombinant polypeptides in bacteria and opened to biology researchers new avenues as obtaining suitable amounts of proteins necessary for their experimental work became easy, fast, and inexpensive. Nevertheless, the recombinant approach remained somehow unpredictable, since many constructs resisted to apparent production or accumulated as aggregates. Several factors and physical/chemical conditions that could improve the accumulation of native-like protein were identified. At the same time, it was acknowledged that the outcome of most of them was erratic and that almost any protein required its own specific optimized set of conditions to achieve its correct folding. The attempt to understand the critical points specific for recombinant protein production missed the goal of setting universally useful protocols, but contributed to the increase of the rate of success by proposing always new empiric combinations. Nevertheless, the results published in the recent literature allow for a better comprehension of some key mechanisms controlling protein production in E. coli and could enable the elaboration of rational methodologies for improving the quantitative and qualitative features of the produced polypeptides. This result will be achieved when the identification of the limiting step that impairs the accomplishment of the native folding for any single construct will become straightforward. This minireview will discuss how factors such as the expression rate, the folding machinery, and the secretion efficiency may impact the final protein yields.
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基于伴侣的程序，以增加大肠杆菌中产生的可溶性重组蛋白的产量。
影响因子：
3.5
作者：
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影响因子：
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6.4
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8.4
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影响因子：
5.7
作者：
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通讯作者：
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