Constitutive production and efficient secretion of soluble full-length streptavidin by an Escherichia coli 'leaky mutant'

Journal of Biotechnology
Jakob Michael MüllerJoe Max Risse

Abstract

Due to its various applications the protein streptavidin is a highly interesting target for heterologous production. This study focuses on different Escherichia coli-based constructs targeting a high-level expression and secretion of streptavidin to the medium. The effect of various promoters, variants of the target gene, leader sequences and host strains on expression and secretion into the culture broth was analyzed. Constitutive production of full-length streptavidin fused with the leader sequence of the bglA gene from Bacillus amyloliquefaciens by the periplasmic 'leaky mutant' E. coli JW1667-5 (Δlpp-752:kan) at 30°C generated the highest yield of the conditions tested, surpassing the extracellular concentration of a conventional T7-based expression system. Supplementation of the medium by the non-ionic surfactants Triton(®) X-100 and X-45 led to an improved secretion of the protein to the culture supernatant. Tetrameric concentrations of streptavidin of 2790±166nM were reached in shake flasks at a productivity of 49.6nMh(-1). Optimization of conditions led to a successful transfer to the bioreactor, yielding a maximal concentration of 2608±169nM and a productivity of 65.2nMh(-1) in fed-batch operation. The proportion of bi...Continue Reading

References

Jan 1, 1978·Annual Review of Biochemistry·J M DiRienzoM Inouye
Jun 27, 1990·Journal of Chromatography·E A Bayer, M Wilchek
Jan 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·T Sano, C R Cantor
Apr 15, 1989·The Biochemical Journal·E A BayerM Wilchek
Feb 25, 1986·Nucleic Acids Research·C E ArgarañaC R Cantor
Oct 6, 1999·Current Opinion in Biotechnology·F Baneyx
May 5, 2000·Biomolecular Engineering·A Skerra, T G Schmidt
Apr 2, 2002·Protein Expression and Purification·Sau-Ching WuSui-Lam Wong
Jul 20, 1964·Archives of Biochemistry and Biophysics·L CHAIET, F J WOLF
Dec 19, 2003·Protein Expression and Purification·Thorsten Lamla, Volker A Erdmann
Feb 18, 2004·Applied Microbiology and Biotechnology·J H Choi, S Y Lee
Feb 17, 2007·Biotechnology and Bioengineering·Ye NiRachel R Chen
Sep 17, 2008·Applied Microbiology and Biotechnology·Gerhard MikschErwin Flaschel
Apr 14, 2009·Nature Methods·Daniel G GibsonHamilton O Smith
Aug 24, 2012·Biotechnology Progress·Marisa C F CasteluberFlávia M L Passos

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Citations

Apr 20, 2016·Biotechnology Progress·Jakob Michael MüllerJoe Max Risse
Jan 19, 2019·Microbial Cell Factories·Charlotte A GreenGraham P Stafford
Aug 28, 2020·Journal of Industrial Microbiology & Biotechnology·Frank K AgbogboJesse McCool
Apr 14, 2018·Engineering in Life Sciences·Gabriele R M Kleiner-GroteKarl Friehs
Jan 25, 2021·Journal of Biotechnology·Jens KastenhoferOliver Spadiut
Feb 2, 2021·Applied Microbiology and Biotechnology·Marina VortmannBruno M Moerschbacher

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