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

Browning, Douglas F., Richards, Kirsty L., Peswani, Amber R., Roobol, Jo, Busby, Stephen J. W., Robinson, Colin (2017) Escherichia coli ‘TatExpress’ strains super-secrete human growth hormone into the bacterial periplasm by the Tat pathway. Biotechnology and Bioengineering, 114 (12). pp. 2828-2836. ISSN 0006-3592. E-ISSN 1097-0290. (doi:10.1002/bit.26434)

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Abstract

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?h 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

Item Type: Article
DOI/Identification number: 10.1002/bit.26434
Subjects: Q Science
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: Colin Robinson
Date Deposited: 06 Sep 2017 07:57 UTC
Last Modified: 23 Jan 2020 04:13 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/63243 (The current URI for this page, for reference purposes)
Robinson, Colin: https://orcid.org/0000-0002-1739-032X
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