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IPTG-independent autoinduction of extracellular matrix proteins using recombinant E. coli as the expression host

Polymer Journal volume 53, pages 385–391 (2021)Cite this article

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Abstract

At present, isopropyl β-D-thiogalactopyranoside (IPTG) is the universal inducer for expressing recombinant proteins under the lac operator/repressor system. In this study, we propose an autoinduction (IPTG-independent) system for recombinant proteins using E. coli as the expression host. We applied this bacterial host for autoinduction to the expression of recombinant proteins, including green fluorescence protein (GFP) and an artificial extracellular matrix protein (aECM-CS5-ELF). The host harbors a mutant Ala294Gly/Thr251Gly phenylalanyl-tRNA synthetase (PheRS**) with an enlarged binding pocket that is expressed under the control of the T7 promoter. Using this system, we demonstrate marked overexpression of the biosynthesized GFP and aECM-CS5-ELF from a 1-L culture containing glucose (5 g/L) and galactose (20 g/L) as the carbon sources, with GFP and aECM-CS5-ELF yields 2.3- and 8.1-fold higher, respectively, than that from an IPTG-induced culture. This unique trial is intended to stimulate novel overexpression strategies based on autoinduction.

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Acknowledgements

AT is grateful to Prof. David A. Tirrell for the opportunity to work in his laboratory at the California Institute of Technology. The authors acknowledge support from the Nagoya Institute of Technology’s Research Promotion Program, from NIH grants EB1971 and GM 62523 and from the NSF Center for the Science and Engineering of Materials at the California Institute of Technology.

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  1. Life and Applied Chemistry, Graduate School of Engineering, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya, 466-8555, Japan

    Kaho Kataoka & Akinori Takasu

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  1. Kaho Kataoka
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  2. Akinori Takasu
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Correspondence to Akinori Takasu.

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Kataoka, K., Takasu, A. IPTG-independent autoinduction of extracellular matrix proteins using recombinant E. coli as the expression host. Polym J 53, 385–391 (2021). https://doi.org/10.1038/s41428-020-00411-9

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