Towards a 'chassis' for bacterial magnetosome biosynthesis: genome streamlining of Magnetospirillum gryphiswaldense by multiple deletions

Microbial Cell Factories

Jörn Kalinowski¹  Christian Rückert¹  Tobias Busche¹  Theresa Zwiener²  Frank Mickoleit²  René Uebe²  Dirk Schüler²  Marina Dziuba³ 

[1]Center for Biotechnology, University of Bielefeld, Bielefeld, Germany

[2]Department of Microbiology, University of Bayreuth, Bayreuth, Germany

[3]Department of Microbiology, University of Bayreuth, Bayreuth, Germany

[4]Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow, Russia

关键词: Magnetospirillum gryphiswaldense;    Magnetotactic bacteria;    Magnetosomes;    Genome reduction;    Chassis;    IS elements;   

DOI  :  10.1186/s12934-021-01517-2

来源: Springer

【 摘 要 】

BackgroundBecause of its tractability and straightforward cultivation, the magnetic bacterium Magnetospirillum gryphiswaldense has emerged as a model for the analysis of magnetosome biosynthesis and bioproduction. However, its future use as platform for synthetic biology and biotechnology will require methods for large-scale genome editing and streamlining.ResultsWe established an approach for combinatory genome reduction and generated a library of strains in which up to 16 regions including large gene clusters, mobile genetic elements and phage-related genes were sequentially removed, equivalent to ~ 227.6 kb and nearly 5.5% of the genome. Finally, the fragmented genomic magnetosome island was replaced by a compact cassette comprising all key magnetosome biosynthetic gene clusters. The prospective 'chassis' revealed wild type-like cell growth and magnetosome biosynthesis under optimal conditions, as well as slightly improved resilience and increased genetic stability.ConclusionWe provide first proof-of-principle for the feasibility of multiple genome reduction and large-scale engineering of magnetotactic bacteria. The library of deletions will be valuable for turning M. gryphiswaldense into a microbial cell factory for synthetic biology and production of magnetic nanoparticles.

【 授权许可】

CC BY