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 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.
journal_name
Biotechnol Bioengjournal_title
Biotechnology and bioengineeringauthors
Browning DF,Richards KL,Peswani AR,Roobol J,Busby SJW,Robinson Cdoi
10.1002/bit.26434subject
Has Abstractpub_date
2017-12-01 00:00:00pages
2828-2836issue
12eissn
0006-3592issn
1097-0290journal_volume
114pub_type
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