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.

中文翻译：

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谷氨酸棒杆菌底盘C1 *：构建和测试用于合成生物学和工业生物技术的新型平台宿主

有针对性的自上而下的基因组减少策略被认为具有为合成生物学和工业生物技术提供强大的基本菌株的巨大潜力。最近，我们创建了一个包含26个基因组还原型谷氨酸棒杆菌菌株的文库，这些菌株在单个基因簇中带有广泛的缺失，并显示出类似野生型的生物适应性。在这里，我们以两个平行顺序对这些不相关的基因簇进行组合删除，并在各种环境条件下对所得的28个菌株进行了文库表征。最终的底盘菌株C1 *携带了13.4％的基因组减少量（412个缺失的基因），并在带有d的确定培养基中显示了类似野生型的生长行为。-葡萄糖作为碳和能源。此外，C1 *被证明可抵抗多种压力（包括氧气限制），并在确定和复杂的培养基条件下显示出长期的生长稳定性。除了提供新颖的原核底盘菌株外，我们的研究结果还包括一个大型菌株文库和一个经过修订的基因组注释列表，这将为谷氨酸棒杆菌的未来系统研究提供有价值的资源。