Magnetosome

Figure 1: Magnetosome

Magnetosome is a kind of rare intracellular membrane-bound structure in a specific type of prokaryotes, of nano-size ranging about 35 - 120 nm. Magnetosomes comprise of a magnetic mineral crystal encapsulated by a lipid bilayer about 3 – 4 nm thick (fig. _1_). A number of common cytoplasmic membrane fatty acids components can be found in the magnetosome membrane, which gives rise to the idea that they are membrane invaginations originated from the cytoplasmic membrane [1].

The magnetosome membrane is highly significant for its biogenesis as it creates an isolated environment within the cell crucial for mineral crystal nucleation and growth [2]. These inorganic crystals are magnetic in nature (hence its name), which compose of either magnetite (Fe₃O₄) or greigite (Fe₃S₄). The magnetosomes usually arrange in one or multiple chains along the cell axis. Different varieties of crystal morphologies such as cubo-octahedral, elongated hexagonal prismatic, and bullet-shaped morphologies were discovered in different magnetotactic bacteria [1].

Magnetotactic Bacteria - The Magnetosome Producer

Figure 2: Micrograph of a Magnetotactic Bacteria, (Species name!!!)

Magnetosomes are organelles synthesized by magnetotactic bacteria for its movement along magnetic field. First discovered in 1975 by Richard Blakemore, these magnetotactic bacteria are mobile, aquatic, gram-negative prokaryotes [3] with an array of cellular morphologies, including coccoid, rod-shaped, vibrioid, helical or even multi-cellular. They are found optimally grown at the oxic-anoxic interface in aquatic habitats, and in fact grow less happily under atmospheric oxygen concentration.

Magnetosomes form a chain and are aligned along the axis within the bacteria. With these magnetosomes inside them, they are able to align passively to the earth’s magnetic field so as to swim along geomagnetic field lines. This behaviour is called magnetotaxis [4] and is beneficial to their survival by aiding them to reach regions of optimal oxygen concentrations at minimal energy cost [5].

Azotobacter vinelandii - What and Why?

Though magnetotactic bacteria is the origin of magnetosomes, these bacteria are described by scientists as a group of fastidious prokaryotic bacteria -- meaning that they are difficult to cultivate owing to their unusual growth requirements. Their micro-aerophilic nature require elaborate growth techniques, and they are difficult to grow on the surface of agar plates, which would make the screening for mutants a problem [6].

The lack of effective methods of DNA transfer in these microorganisms is a challenge too. Luckily, the situation is improving due to better technologies recently and some of the genes from M. magnetotacticum have been confirmed functionally expressed in E. coli. This shows that the transcriptional and translational elements of the two microorganisms are compatible. With such good news, a number of previous igem teams including kyoto: 2014, OCU-China: 2013, Washington: 2011 and UNIK_Copenhagen: 2013 have been working with transferring the magnetosome related genes to e.coli. Some exciting results the formation of magnetosome membrane in e.coli (by the kyoto team) has been reported by previous teams, however, never the whole magnetosome.

Figure 3: Azotobacter vinelandii

We have been wondering why magnetosomes seems so hard to be formed in e.coli. And then, we come up with a hypothesis -- the formation of magnetosome requires a micro-aerobic or anaerobic environment as the magnetotatic bacteria are all living micro-aerobically.

Therefore, we chose a new bacteria to work on our magnetosome project -- the Azotobacter vinelandii.

Azotobacter vinelandii is gram-negative diazotroph. It is a soil bacterium related to the Pseudomonas genus that fixes nitrogen under aerobic conditions while having enzymatic mechanisms protecting its oxygen-sensitive nitrogenase from oxygen damage. This findings shows that A. vinelandii is an excellent host for the production and characterization of oxygen-sensitive proteins or organelles as in our case [7].

With the biggest advantage of using azotobacter is that it is an aerobic bacteria with an intracellular anaerobic condition, we can grow it easily in normal conditions in lab without expensive equipments like fermenter, while fulfilling the growing criteria for the magnetosome. Besides, most parts in registry are functionable in Azotobacter and Azotobacter is of safety level group 1 too. One more important thing is that it can do homologous recombination by itself which is a critical process we need in our project.