Ukkonen Kaisa, Neubauer Antje, Pereira Vinit J, Vasala Antti
BioSilta Oy, Aapistie 32, Oulu, 90220, Finland.
Enpresso GmbH, Berlin, Germany.
Methods Mol Biol. 2017;1586:127-137. doi: 10.1007/978-1-4939-6887-9_8.
Expression of recombinant proteins in sufficient quantities is essential for protein structure-function studies. The most commonly used method for recombinant protein production is overexpression in E. coli cultures. However, producing high yields of functional proteins in E. coli can be a challenge in conventional shaken cultures. This is often due to nonoptimal growth conditions, which result in low cell yields and insoluble or incorrectly folded target protein. To overcome the shortcomings of shake flask cultivation, we present a cultivation method based on enzymatic glucose delivery. This system mimics the fed-batch principle used in bioreactor cultivations and provides high yields of biomass and recombinant proteins in shaken cultivations.
大量表达重组蛋白对于蛋白质结构-功能研究至关重要。生产重组蛋白最常用的方法是在大肠杆菌培养物中进行过表达。然而，在传统的摇瓶培养中，要在大肠杆菌中高产功能性蛋白可能具有挑战性。这通常是由于生长条件不理想，导致细胞产量低以及目标蛋白不溶或折叠错误。为了克服摇瓶培养的缺点，我们提出了一种基于酶促葡萄糖递送的培养方法。该系统模拟了生物反应器培养中使用的补料分批培养原理，并在摇瓶培养中提供了高产的生物量和重组蛋白。
