Gram-scale Production of Recombinant Microbial Enzymes in Shake Flasks


Chrast, L., Chaloupkova, R., Damborsky, J.




Heterologous production of recombinant proteins is a cornerstone of microbiological and biochemical research as well as various biotechnological processes. Yields and quality of produced proteins have a tremendous impact on structural and enzymology studies, development of new biopharmaceuticals and establishing new biocatalytic processes. Majority of current protocols for recombinant protein expression in Escherichia coli exploit batch cultures with complex media, often providing low yields of the target protein due to oxygen transfer limitation, rapid depletion of carbon sources, and pH changes during the cultivation. Recently introduced EnBase technology enables fed-batch-like cultivations in shake flasks with continuous glucose release from a soluble starch. In this study, we critically compare the yields of fourteen model enzymes in E. coli cultured in a novel semi-defined medium and in a complex medium. Significant improvements in the volumetric yields 2-31 times were observed for all tested enzymes expressed in enzymatic fed-batch-like cultures with no adverse impact on enzyme structure, stability, or activity. Exceptional yields, higher than one gram of protein per litre of culture, were obtained with six enzymes. We conclude that the novel semi-defined medium tested in this study provides a robust improvement of protein yields in shake flasks without investment into costly bioreactors.

Full text


Chrast, L., Chaloupkova, R., Damborsky, J., 2018: Gram-scale Production of Recombinant Microbial Enzymes in Shake Flasks. FEMS Microbiology Letters 365: fnx265.

Gaspar a Gaussian Expert
1st Hands-on Computational Enzyme Design Workshop
Marie Curie Fellowship for Dmitri
Rayyan Announced as Best Presenter
Researchers’ Night 2019 in LL
Open Postdoctoral Position: Protein Structural Biologist
Andrea Awarded for Excellent Research