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

Zwiener, Theresa; Dziuba, Marina; Mickoleit, Frank; Rückert, Christian; Busche, Tobias; Kalinowski, Jörn; Uebe, René; Schüler, Dirk

doi:10.1186/s12934-021-01517-2

Cited by 18 publications

(9 citation statements)

References 93 publications

(80 reference statements)

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“…Genome minimization has also been performed in non-model organisms to produce various biomolecules. For example, Magnetospirillum gryphiswaldense is a key organism in magnetosome biosynthesis and production ( Zwiener et al, 2021 ). Magnetosomes have biological functions as magnetic sensors and have strong biomedical and biotechnological applications as magnetic nanoparticles in magnetic imaging, carriers for magnetic drug targeting and hyperthermia applications.…”

Section: Applications Of Reduced-genome Bacterial Strainsmentioning

confidence: 99%

“…Magnetosomes have biological functions as magnetic sensors and have strong biomedical and biotechnological applications as magnetic nanoparticles in magnetic imaging, carriers for magnetic drug targeting and hyperthermia applications. Since large scale production is limiting its applicability, the genome of M. gryphiswaldense has been reduced by 5.5% to simplify its metabolism and work towards a chassis for magnetosome production ( Zwiener et al, 2021 ). This strain displayed similar growth rates and magnetosome biosynthesis as the parental strain but had increased genetic stability and resilience ( Zwiener et al, 2021 ).…”

Section: Applications Of Reduced-genome Bacterial Strainsmentioning

confidence: 99%

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Bacterial genome reductions: Tools, applications, and challenges

LeBlanc

Charles

2022

Front. Genome Ed.

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Bacterial cells are widely used to produce value-added products due to their versatility, ease of manipulation, and the abundance of genome engineering tools. However, the efficiency of producing these desired biomolecules is often hindered by the cells’ own metabolism, genetic instability, and the toxicity of the product. To overcome these challenges, genome reductions have been performed, making strains with the potential of serving as chassis for downstream applications. Here we review the current technologies that enable the design and construction of such reduced-genome bacteria as well as the challenges that limit their assembly and applicability. While genomic reductions have shown improvement of many cellular characteristics, a major challenge still exists in constructing these cells efficiently and rapidly. Computational tools have been created in attempts at minimizing the time needed to design these organisms, but gaps still exist in modelling these reductions in silico. Genomic reductions are a promising avenue for improving the production of value-added products, constructing chassis cells, and for uncovering cellular function but are currently limited by their time-consuming construction methods. With improvements to and the creation of novel genome editing tools and in silico models, these approaches could be combined to expedite this process and create more streamlined and efficient cell factories.

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Section: Applications Of Reduced-genome Bacterial Strainsmentioning

confidence: 99%

Section: Applications Of Reduced-genome Bacterial Strainsmentioning

confidence: 99%

Bacterial genome reductions: Tools, applications, and challenges

LeBlanc

Charles

2022

Front. Genome Ed.

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“…BAS is one of the most available tools for biological research, used in protein purification, protein localization analysis, and immobilization [ 46 , 47 , 48 , 49 ]. In this study, we adopted the BMP surface display technique to construct biotinylated BMPs in Magnetospirillum gryphiswaldense , the model strain for analyzing BMPs synthesis and bioproduction [ 50 ]. Moreover, we attempted to increase the throughput of engineered BMPs.…”

Section: Introductionmentioning

confidence: 99%

Biomanufacturing Biotinylated Magnetic Nanomaterial via Construction and Fermentation of Genetically Engineered Magnetotactic Bacteria

Zheng

et al. 2022

Bioengineering

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Biosynthesis provides a critical way to deal with global sustainability issues and has recently drawn increased attention. However, modifying biosynthesized magnetic nanoparticles by extraction is challenging, limiting its applications. Magnetotactic bacteria (MTB) synthesize single-domain magnetite nanocrystals in their organelles, magnetosomes (BMPs), which are excellent biomaterials that can be biologically modified by genetic engineering. Therefore, this study successfully constructed in vivo biotinylated BMPs in the MTB Magnetospirillum gryphiswaldense by fusing biotin carboxyl carrier protein (BCCP) with membrane protein MamF of BMPs. The engineered strain (MSR−∆F−BF) grew well and synthesized small-sized (20 ± 4.5 nm) BMPs and were cultured in a 42 L fermenter; the yield (dry weight) of cells and BMPs reached 8.14 g/L and 134.44 mg/L, respectively, approximately three-fold more than previously reported engineered strains and BMPs. The genetically engineered BMPs (BMP−∆F−BF) were successfully linked with streptavidin or streptavidin-labelled horseradish peroxidase and displayed better storage stability compared with chemically constructed biotinylated BMPs. This study systematically demonstrated the biosynthesis of engineered magnetic nanoparticles, including its construction, characterization, and production and detection based on MTB. Our findings provide insights into biomanufacturing multiple functional magnetic nanomaterials.

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“…Since first reported in 1975 [1] magnetotactic bacteria (MTB) have attracted interest because of their abilities to synthesize magnetite crystals in specialized organelles called "magnetosomes" [2][3][4]. Gene regulation and genomic analysis related to magnetosome formation has been extensively studied [5][6][7]. Superparamagnetic magnetite crystals of similar size and shape to the bacterial magnetites are formed in vitro due to the presence of recombinant Mms6, a magnetosome-associated protein [8][9][10].…”

Section: Introductionmentioning

confidence: 99%

The Magnetosome Protein, Mms6, is a Lipid-Activated Ferric Reductase

Singappuli-Arachchige¹,

Feng²,

Wang³

et al. 2022

Preprint

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Magnetosomes of magnetotactic bacteria consist of magnetic nanocrystals with defined morphologies enclosed in vesicles originated from cytoplasmic membrane invaginations. Although many proteins are involved in creating magnetosomes, a single magnetosome protein, Mms6, can direct the crystallization of magnetite nanoparticles in vitro. The in vivo role of Mms6 in magnetosome formation is debated and the observation that Mms6 binds ferric and not ferrous iron raises the question of how Mms6 could promote the crystallization of magnetite, which contains both ferric and ferrous iron. Here we show that Mms6 is a ferric reductase that reduces ferric to ferrous iron using NADH and FAD as electron donor and cofactor, respectively. Reductase activity is elevated when Mms6 is integrated into either liposomes or bicelles. Analysis of Mms6 mutants suggests that the C-terminal domain binds iron and the N-terminal domain contains the catalytic site. Although Mms6 forms multimers that involve C-terminal and N-terminal domain interactions, a fusion protein with Mms6, which remains a monomer, displays reductase activity, which suggests that the catalytic site is fully in the monomer. These results are consistent with a hypothesis that Mms6, a membrane protein, promotes the formation of magnetite by a mechanism that involves reducing iron.

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

Cited by 18 publications

References 93 publications

Bacterial genome reductions: Tools, applications, and challenges

Bacterial genome reductions: Tools, applications, and challenges

Biomanufacturing Biotinylated Magnetic Nanomaterial via Construction and Fermentation of Genetically Engineered Magnetotactic Bacteria

The Magnetosome Protein, Mms6, is a Lipid-Activated Ferric Reductase

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