Article Open Access

Time-efficient and Semi-automated Production and Screening of Proteins

Synthetic Biology and Engineering. 2023, 1(3), 10016;
S. R. Sekar 1,    S. Ilhan 1,    Uwe Jandt 1,    An-Ping Zeng 1, 2, *   
Institute of Bioprocess and Biosystems Engineering, Hamburg University of Technology, Denickestr. 15, Hamburg 21073, Germany
Center of Synthetic Biology and Integrated Bioengineering, School of Engineering, Westlake University, Hangzhou 310024, China
Authors to whom correspondence should be addressed.

Received: 20 Nov 2023    Accepted: 13 Dec 2023    Published: 18 Dec 2023   


Fast, flexible and non-randomized modification, production and screening of proteins in fully automated system are of high interest in biological research and applications. The conventional methods for protein engineering and screening, especially for mutations of multiple residues. are time consuming and often unreliable. We demonstrate here a new, fast and flexible protein production and screening method which combines linear expression template (LET) based cell free protein synthesis (CFPS) with specific screening methods. This approach is demonstrated using green fluorescence protein, phosphoserine aminotransferase (serC) and aspartokinase III (AKIII) as model systems. The results show that mutants with changes in different protein properties upon multiple point mutations can be produced and screened within 6 to 15 h. This method can be used further to generate mutants of enzymes and multi-enzyme complexes and be implemented within the workflow of a feedback-guided protein optimization and screening system.


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