Corynebacterium glutamicum Chassis C1*: Building and Testing a Novel Platform Host for Synthetic Biology and Industrial Biotechnology

5 years ago

Corynebacterium glutamicum Chassis C1*: Building and Testing a Novel Platform Host for Synthetic Biology and Industrial Biotechnology

Volker F. Wendisch, Christian Rückert, Jörn Kalinowski, Moritz Fabian Müller, Julia Frunzke, Natalie Brühl, Andreas Küberl, Stephan Hans, Daniel Siebert, Michael Bott, Kay Marin, Meike Baumgart, Reinhard Krämer, Gerd Seibold, Andreas Radek, Ramona Kloß, Tino Polen, Simon Unthan, Wolfgang Wiechert, Niklas Tenhaef, Stephan Noack

Targeted top-down strategies for genome reduction are considered to have a high potential for providing robust basic strains for synthetic biology and industrial biotechnology. Recently, we created a library of 26 genome-reduced strains of Corynebacterium glutamicum carrying broad deletions in single gene clusters and showing wild-type-like biological fitness. Here, we proceeded with combinatorial deletions of these irrelevant gene clusters in two parallel orders, and the resulting library of 28 strains was characterized under various environmental conditions. The final chassis strain C1* carries a genome reduction of 13.4% (412 deleted genes) and shows wild-type-like growth behavior in defined medium with d-glucose as carbon and energy source. Moreover, C1* proves to be robust against several stresses (including oxygen limitation) and shows long-term growth stability under defined and complex medium conditions. In addition to providing a novel prokaryotic chassis strain, our results comprise a large strain library and a revised genome annotation list, which will be valuable sources for future systemic studies of C. glutamicum.

Publisher URL: http://dx.doi.org/10.1021/acssynbio.7b00261

DOI: 10.1021/acssynbio.7b00261