DisCoTune: versatile auxiliary plasmids for the production of disulphide-containing proteins and peptides in the E. coli T7 system
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DisCoTune : versatile auxiliary plasmids for the production of disulphide-containing proteins and peptides in the E. coli T7 system. / Bertelsen, Andreas B.; Hackney, Celeste Menuet; Bayer, Carolyn N.; Kjelgaard, Lau D.; Rennig, Maja; Christensen, Brian; Sørensen, Esben Skipper; Safavi-Hemami, Helena; Wulff, Tune; Ellgaard, Lars; Nørholm, Morten H. H.
I: Microbial Biotechnology, Bind 14, Nr. 6, 2021, s. 2566-2580.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - DisCoTune
T2 - versatile auxiliary plasmids for the production of disulphide-containing proteins and peptides in the E. coli T7 system
AU - Bertelsen, Andreas B.
AU - Hackney, Celeste Menuet
AU - Bayer, Carolyn N.
AU - Kjelgaard, Lau D.
AU - Rennig, Maja
AU - Christensen, Brian
AU - Sørensen, Esben Skipper
AU - Safavi-Hemami, Helena
AU - Wulff, Tune
AU - Ellgaard, Lars
AU - Nørholm, Morten H. H.
N1 - Publisher Copyright: © 2021 The Authors. Microbial Biotechnology published by Society for Applied Microbiology and John Wiley & Sons Ltd.
PY - 2021
Y1 - 2021
N2 - Secreted proteins and peptides hold large potential both as therapeutics and as enzyme catalysts in biotechnology. The high stability of many secreted proteins helps maintain functional integrity in changing chemical environments and is a contributing factor to their commercial potential. Disulphide bonds constitute an important post-translational modification that stabilizes many of these proteins and thus preserves the active state under chemically stressful conditions. Despite their importance, the discovery and applications within this group of proteins and peptides are limited by the availability of synthetic biology tools and heterologous production systems that allow for efficient formation of disulphide bonds. Here, we refine the design of two DisCoTune (Disulphide bond formation in E. coli with tunable expression) plasmids that enable the formation of disulphides in the highly popular Escherichia coli T7 protein production system. We show that this new system promotes significantly higher yield and activity of an industrial protease and a conotoxin, which belongs to a group of disulphide-rich venom peptides from cone snails with strong potential as research tools and pharmacological agents.
AB - Secreted proteins and peptides hold large potential both as therapeutics and as enzyme catalysts in biotechnology. The high stability of many secreted proteins helps maintain functional integrity in changing chemical environments and is a contributing factor to their commercial potential. Disulphide bonds constitute an important post-translational modification that stabilizes many of these proteins and thus preserves the active state under chemically stressful conditions. Despite their importance, the discovery and applications within this group of proteins and peptides are limited by the availability of synthetic biology tools and heterologous production systems that allow for efficient formation of disulphide bonds. Here, we refine the design of two DisCoTune (Disulphide bond formation in E. coli with tunable expression) plasmids that enable the formation of disulphides in the highly popular Escherichia coli T7 protein production system. We show that this new system promotes significantly higher yield and activity of an industrial protease and a conotoxin, which belongs to a group of disulphide-rich venom peptides from cone snails with strong potential as research tools and pharmacological agents.
U2 - 10.1111/1751-7915.13895
DO - 10.1111/1751-7915.13895
M3 - Journal article
C2 - 34405535
AN - SCOPUS:85112730171
VL - 14
SP - 2566
EP - 2580
JO - Microbial Biotechnology
JF - Microbial Biotechnology
SN - 1751-7907
IS - 6
ER -
ID: 278493991