Biomineralisation of Magnetosomes in Bacteria Microbial Bionanotechnology Chapter 5

2 Magnetotactic Magnetosome – Crystalline particles of iron oxide or sulfide – Magnetite Fe 3 O 4 – Greigite Fe 3 S 4 All are either obligate microaerophiles or strict anaerobes Motile, aquatic bacteria Direction of motility is affected by the Earth’s geomagnetic field Strains are either north- or south-seeking depending upon oxic conditions – North-seekers predominate in the northern hemisphere – South-seekers predominate the southern hemisphere – Exist in equal numbers at the equator Current hypothesis states that these bacteria use the geomagnetic field to locate lower O 2 or anaerobic habitats

Types of magnetotaxis

Types of magneto-aerotaxis

5 There are Two Types of Magneto- aerotaxis... Axial Magneto- aerotaxis Polar Magneto- aerotaxis e.g., Magnetospirillum magnetotacticum e.g., strain MC-1, a magnetotactic coccus

Biomineralisation of Fe

Magnetotactic Bacteria Electron cryotomography of Magnetospirillum magneticum sp. AMB-1 reveals that magnetosomes are invaginations of the inner membrane. (A) General features of AMB-1 cells highlighted in a 12-nm-thick section of an ECT reconstruction. Outer membrane, OM; inner membrane, IM; peptidoglycan layer, PG; ribosomes, R; outer membrane bleb, B; chemoreceptor bundle, CR; poly-ß-hydroxybutyrate granule, PHB; gold fiduciary marker, G; magnetosome chain, MG. Scale bar, 500 nm. (B to E) Representative magnetosomes containing no magnetite (B), small (C), mediumsized (D), and fully-grown (E) crystals are invaginations of the inner membrane. Scale bar, 50 nm.

Magnetosome chains are flanked by long cytoskeletal filaments. (A) Larger view of the magnetosome chain in Fig. 1A. (B) Similar view of a magnetosome chain grown in the absence of iron, which prevents the formation of magnetite crystals. Arrows point to the long filaments. (C) Three-dimensional organization of magnetosomes (yellow) and their associated filaments (green) shown in (B) with respect to the whole cell (blue). Scale bars, 100 nm.Fig. 1A

Bakterielle Magnetosomer TEM image of two Itaipu-1 cocci. Each bacterium has two chains of magnetosomes (arrows) and two phosphorus-rich globules (P). Scale bar, 1 μm.

High-resolution TEM images of Itaipu-1 magnetosomes with indexed bars parallel to lattice planes. Obvious symmetries between even the very small facets on opposite sides of the crystal diagonals can be seen. Comparison with other crystals of the chain in Fig. 2 also indicates that this symmetry regularly alternates between crystals.2

(A) Electrostatic contribution to the holographic phase shift from the Itaipu magnetosomes shown in Fig. 2, oriented to a [110] projection. The contours represent the projected thickness and show a flat-topped morphology and steep sides. (B) Projected thickness contours for the same crystals after tilting by 30° about the chain axis to a [211] orientation. The contours show that the crystal is much thicker along its center than along its edges, having a central ridge formed by intersecting faces. (C) Line profiles (solid line for panel A and dashed line for panel B) across the magnetosomes from the indicated positions (arrows), converted to values of one- half their thickness, reveal a 120° angle between the facets for the [211] projection, which is consistent with the intersection of [110] faces. Scale bar, 150 nm (panels A and B).2

TEM images of Itaipu-1 and Itaipu-3 magnetosomes. (A) Chain of large magnetosomes from magnetotactic bacterial strain Itaipu-1 surrounded by smaller, elongated magnetosomes from strain Itaipu-3. The inset is a [211] diffraction pattern from the second large Itaipu-1 crystal (arrow). (B) Same chain as in panel A tilted 30 o about the [111] axis. The inset [110] diffraction pattern from the second large Itaipu-1 crystal shows (111) fringes from the magnetically easy axis. Corner faces {111} and {200} are mirrored about the vertical (or horizontal) axis for alternating crystals (double arrows); see detailed image in Fig. 3. Scale bar, 200 nm.3

Tomographic reconstruction of a magnetite nanocrystal from an undescribed coccus collected from Sweet Springs Nature Reserve, Morro Bay, CA, reconstructed from a tilt series of STEM HAADF images obtained at 300 kV on a Philips CM300 FEG TEM over a range of ± 56°. The tableau shows the three-dimensional morphology of the crystal viewed from a range of directions.

Magnetosome Element Analysis

Cu-Fe Analysis

Magnetosomes

Magnetosome crystal morphology

MamK, a homolog of the bacterial actin-like protein MreB, forms filaments in vivo. (A) Phylogenetic relationship between MamK and other bacterial actin-like proteins demonstrated by an unrooted tree. These proteins separate into three distinct groups: MamK (green), ParM/StbA (red) and MreB (blue). (B) MamK fused to GFP (green) appears to form filaments in vivo localized to the inner curvature of the cell (cell membrane stained red with FM4-64).

MamK is required for the proper organization of the magnetosome chain. (A) Three-dimensional reconstruction of a wild-type AMB-1 cell. The cell membrane (gray), magnetosome membrane (yellow), magnetite (orange), and magnetosome-associated filaments (green) are rendered. (B) mamK mutant, where magnetosomes appear disordered and no filaments are found in their vicinity. (C) mamK cell expressing mamK-GFP on a plasmid showing full reversal of the mutant phenotype.

Magnetosome membrane proteins

Comparative genome analysis

Magnetotaxis genes

Mam gene regulation in Cells

Magnetosome membrane

Magnetosome membrane protein

Biotechnological applications Delivery systems Separation systems DNA arrays RNA arrays Thermo treatment Sensor systems

Applications of Magnetosome particles