Expression and purification of ELP-intein-tagged target proteins in high cell density E. coli fermentationReport as inadecuate




Expression and purification of ELP-intein-tagged target proteins in high cell density E. coli fermentation - Download this document for free, or read online. Document in PDF available to download.

Microbial Cell Factories

, 9:77

First Online: 19 October 2010Received: 16 August 2010Accepted: 19 October 2010

Abstract

BackgroundElastin-like polypeptides ELPs are useful tools that can be used to non-chromatographically purify proteins. When paired with self-cleaving inteins, they can be used as economical self-cleaving purification tags. However, ELPs and ELP-tagged target proteins have been traditionally expressed using highly enriched media in shake flask cultures, which are generally not amenable to scale-up.

ResultsIn this work, we describe the high cell-density expression of self-cleaving ELP-tagged targets in a supplemented minimal medium at a 2.5 liter fermentation scale, with increased yields and purity compared to traditional shake flask cultures. This demonstration of ELP expression in supplemented minimal media is juxtaposed to previous expression of ELP tags in extract-based rich media. We also describe several sets of fed-batch conditions and their impact on ELP expression and growth medium cost.

ConclusionsBy using fed batch E. coli fermentation at high cell density, ELP-intein-tagged proteins can be expressed and purified at high yield with low cost. Further, the impact of media components and fermentation design can significantly impact the overall process cost, particularly at large scale. This work thus demonstrates an important advances in the scale up of self-cleaving ELP tag-mediated processes.

Electronic supplementary materialThe online version of this article doi:10.1186-1475-2859-9-77 contains supplementary material, which is available to authorized users.

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Author: Baley A Fong - David W Wood

Source: https://link.springer.com/







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