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旧 PET 换新招：通过大肠杆菌混合进料系统对包涵体形成和性质进行调优。

Teaching an old pET new tricks: tuning of inclusion body formation and properties by a mixed feed system in E. coli.

机构信息

Research Division Biochemical Engineering, Institute of Chemical, Environmental and Biological Engineering, TU Wien, Vienna, Austria.

Christian Doppler Laboratory for Mechanistic and Physiological Methods for Improved Bioprocesses, Institute of Chemical, Environmental and Biological Engineering, TU Wien, Vienna, Austria.

出版信息

Appl Microbiol Biotechnol. 2018 Jan;102(2):667-676. doi: 10.1007/s00253-017-8641-6. Epub 2017 Nov 20.

DOI:10.1007/s00253-017-8641-6
PMID:29159587
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5756567/
Abstract

Against the outdated belief that inclusion bodies (IBs) in Escherichia coli are only inactive aggregates of misfolded protein, and thus should be avoided during recombinant protein production, numerous biopharmaceutically important proteins are currently produced as IBs. To obtain correctly folded, soluble product, IBs have to be processed, namely, harvested, solubilized, and refolded. Several years ago, it was discovered that, depending on cultivation conditions and protein properties, IBs contain partially correctly folded protein structures, which makes IB processing more efficient. Here, we present a method of tailored induction of recombinant protein production in E. coli by a mixed feed system using glucose and lactose and its impact on IB formation. Our method allows tuning of IB amount, IB size, size distribution, and purity, which does not only facilitate IB processing, but is also crucial for potential direct applications of IBs as nanomaterials and biomaterials in regenerative medicine.

摘要

针对包涵体（IBs）在大肠杆菌中仅为错误折叠蛋白质的无活性聚集体的过时观点，目前有许多具有生物制药重要性的蛋白质被作为 IBs 进行生产。为了获得正确折叠、可溶性的产物，必须对 IBs 进行处理，即收获、溶解和重折叠。几年前，人们发现，根据培养条件和蛋白质特性，IBs 中含有部分正确折叠的蛋白质结构，这使得 IB 处理更加高效。在这里，我们介绍了一种通过使用葡萄糖和乳糖的混合进料系统来定制诱导大肠杆菌中重组蛋白生产的方法，以及其对 IB 形成的影响。我们的方法允许调整 IB 的数量、大小、大小分布和纯度，这不仅有助于 IB 的处理，而且对于将 IBs 作为纳米材料和生物材料直接应用于再生医学领域也是至关重要的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/288b/5756567/f342ce7bee76/253_2017_8641_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/288b/5756567/ae0e52db8534/253_2017_8641_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/288b/5756567/a4429fc2eb55/253_2017_8641_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/288b/5756567/3329dd615121/253_2017_8641_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/288b/5756567/3a872939995f/253_2017_8641_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/288b/5756567/f342ce7bee76/253_2017_8641_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/288b/5756567/ae0e52db8534/253_2017_8641_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/288b/5756567/a4429fc2eb55/253_2017_8641_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/288b/5756567/3329dd615121/253_2017_8641_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/288b/5756567/3a872939995f/253_2017_8641_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/288b/5756567/f342ce7bee76/253_2017_8641_Fig5_HTML.jpg

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