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

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Zwiener, Theresa ; Dziuba, Marina ; Mickoleit, Frank ; Rückert, Christian ; Busche, Tobias ; Kalinowski, Jörn ; Uebe, René ; Schüler, Dirk:
Towards a "chassis" for bacterial magnetosome biosynthesis : genome streamlining of Magnetospirillum gryphiswaldense by multiple deletions.
In: Microbial Cell Factories. Bd. 20 (2021) . - 35.
ISSN 1475-2859
DOI: https://doi.org/10.1186/s12934-021-01517-2

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Abstract

Abstract
Background: Because 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.
Results: We 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.
Conclusion: We 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.