Escherichia coli "TatExpress" strains super-secrete human growth hormone into the bacterial periplasm by the Tat pathway


Numerous high-value proteins are secreted into the Escherichia coli periplasm by the General Secretory (Sec) pathway, but Sec-based production chassis cannot handle many potential target proteins. The Tat pathway offers a promising alternative because it transports fully folded proteins; however, yields have been too low for commercial use. To facilitate Tat export, we have engineered the TatExpress series of super-secreting strains by introducing the strong inducible bacterial promoter, ptac, upstream of the chromosomal tatABCD operon, to drive its expression in E. coli strains commonly used by industry (e.g., W3110 and BL21). This modification significantly improves the Tat-dependent secretion of human growth hormone (hGH) into the bacterial periplasm, to the extent that secreted hGH is the dominant periplasmic protein after only 1 hr induction. TatExpress strains accumulate in excess of 30 mg L-1 periplasmic recombinant hGH, even in shake flask cultures. A second target protein, an scFv, is also shown to be exported at much higher rates in TatExpress strains.

Publication DOI:
Divisions: College of Health & Life Sciences > School of Biosciences
Additional Information: © 2017 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. Funding: Biotechnology and Biological Sciences Research Council. Grant Number: BB/M018288/1
Uncontrolled Keywords: Escherichia coli/genetics,Gene Products, tat/genetics,Genetic Enhancement/methods,Growth Hormone/biosynthesis,Humans,Metabolic Networks and Pathways/genetics,Periplasm/metabolism,Promoter Regions, Genetic/genetics,Recombinant Proteins/biosynthesis,Secretory Pathway/genetics
Publication ISSN: 1097-0290
Last Modified: 27 Jun 2024 11:02
Date Deposited: 18 Jan 2022 12:55
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Related URLs: https://onlinel ... .1002/bit.26434 (Publisher URL)
PURE Output Type: Article
Published Date: 2017-12
Published Online Date: 2017-08-26
Accepted Date: 2017-08-25
Authors: Browning, Douglas F (ORCID Profile 0000-0003-4672-3514)
Richards, Kirsty L
Peswani, Amber R
Roobol, Jo
Busby, Stephen J W
Robinson, Colin



Version: Published Version

License: Creative Commons Attribution

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