Baumgart Meike, Unthan Simon, Kloß Ramona, Radek Andreas, Polen Tino, Tenhaef Niklas, Müller Moritz Fabian, Küberl Andreas, Siebert Daniel, Brühl Natalie, Marin Kay, Hans Stephan, Krämer Reinhard, Bott Michael, Kalinowski Jörn, Wiechert Wolfgang, Seibold Gerd, Frunzke Julia, Rückert Christian, Wendisch Volker F, Noack Stephan
Institute of Bio- and Geosciences, IBG-1: Biotechnology, Systemic Microbiology, Forschungszentrum Jülich , 52425 Jülich, Germany.
Institute of Bio- and Geosciences, IBG-1: Biotechnology, Systems Biotechnology, Forschungszentrum Jülich , 52425 Jülich, Germany.
ACS Synth Biol. 2018 Jan 19;7(1):132-144. doi: 10.1021/acssynbio.7b00261. Epub 2017 Aug 30.
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.
用于基因组精简的靶向自上而下策略被认为具有很大潜力，可为合成生物学和工业生物技术提供稳健的基础菌株。最近，我们构建了一个包含26株谷氨酸棒杆菌基因组精简菌株的文库，这些菌株在单个基因簇中存在广泛缺失，且具有类似野生型的生物学适应性。在此，我们以两种平行顺序对这些无关基因簇进行组合缺失，并在各种环境条件下对由此产生的28株菌株文库进行了表征。最终的底盘菌株C1的基因组减少了13.4%（412个基因被删除），在以d-葡萄糖作为碳源和能源的限定培养基中表现出类似野生型的生长行为。此外，C1被证明对多种胁迫（包括氧气限制）具有抗性，并且在限定和复杂培养基条件下表现出长期生长稳定性。除了提供一种新型原核底盘菌株外，我们的研究结果还包括一个大型菌株文库和一份修订后的基因组注释列表，这将成为未来谷氨酸棒杆菌系统研究的宝贵资源。
