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Proinsulin was expressed by Escherichia coli SHuffle T7 with pET system in minimal medium containing glucose (Glu medium), glucose and glycerol (GluGly medium) or glycerol (Gly medium).

With 100 μM IPTG, proinsulin production did not increase with glycerol. In contrast, proinsulin production per medium volume in GluGly and Gly media was approximately 3∼4-fold higher than in Glu medium with 10 μM IPTG. mRNA expression of target protein was higher in GluGly versus Glu medium, indicating that proinsulin production was enhanced by the release of glucose-induced catabolite inhibition.

Although proinsulin production did not differ between GluGly and Gly media at 25 hours, substrate was consumed quickly in GluGly medium with 1.55±0.12 times higher proinsulin production at 15 hours. Productivity, considering the production period, was highest in the GluGly medium.

This study shows a mixture of glucose and glycerol is valuable for protein production in E. coli with low IPTG concentrations.

Paper information

Journal: Biotechnology Reports
Title: Induction conditions that promote the effect of glycerol on recombinant protein production in Escherichia coli
DOI: 10.1016/j.btre.2025.e00898
Authors: Yoshihiro Ojima, Hajime Saito, Shintaro Miyuki, Koichi Fukunaga, Terumichi Tsuboi, Masayuki Azuma
Published:  16 May 2025
URL: https://doi.org/10.1016/j.btre.2025.e00898

Contact

Yoshihiro Ojima
Graduate School of Engineering
Email: ojima[at]omu.ac.jp

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