Specifications:

S-Layer: Hexagonal

Lattice Spacing: 19 nm

Pore size: 5.5 nm

• S-layer protein is cross-linked; it cannot be disassembled or reassembled.
• Works well on glass and silicon

Deinococcus radiodurans

Details:

Brightfield micrograph of
D. radiodurans cells

Deinococcus radiodurans is an extraordinarily hardy Gram-positive bacterium that forms reddish-pink colonies. It is the most radiation resistant bacterium known. The relatively large pore size and lattice spacing make S-Layers from this organism ideal for uses where small scale (on the order of ~ 1 µm) patterning is needed.

The S-Layer proteins from D. radiodurans are chemically cross-linked, so that the extracted S-Layers can not be disassembled and reassembled from individual subunit proteins.

Deinococcus radiodurans

Applications:

Generation of hexagonal arrays of patterned metal, metal oxide, semiconductor, or other inorganic nanoclusters

Patterning of biological macromolecules such as enzymes, protein receptors, or nucleic acids

 

>Physical and thermal stabilization of lipid-based structures, such as membrane monolayers, bilayers, and vesicles

Reassembly Conditions:

Deinococcus radiodurans S-Layers are cross-linked and cannot be disassembled or reassembled.

References:

Baumeister, W., Barth, M., Hegerl, R., Guckenberger, R., Hahn, M. and Saxton, W.O. Three-dimensional Structure of the Regular Surface Layer (HPI Layer) of Deinococcus radiodurans. J. Mol. Biol. 187: 241-250.

Peters, J., Peters, M., Lottspeich, F., Schafer, W., and Baumeister, W. (1987) Nucleotide Sequence Analysis of the Gene Encoding the Deinococcus radiodurans Surface Protein, Derived Amino Acid Sequence, and Complementary Protein Chemical Studies. J. Bacteriol. 169: 5216-5233.

Specifications:

S-Layer: Square

Lattice Spacing: 12.5 nm

Pore size: 5.5 nm

• S-layer protein is not cross-linked; it can be disassembled and reassembled.
• Works well on glass, silicon, and will readily reassemble on lipid membranes.

Bacillus sphaericus

Details:

Brightfield micrograph of
B. sphaericus

Bacillus sphaericus is a Gram-positive spore-forming aerobic bacterium, common to soil and aquatic environments. It has been studied for its ability to produce a toxin active against larvae of some mosquitoes.

B. sphaericus produces a square patterned S-layer that can be disassembled into monomers using chaotropic agents such as guanidine HCl. B. sphaericus S-Layers are among the best characterized of the S-Layer proteins. B. sphaericus S-layer monomers will reassemble upon the addition of 10 mM CaCl_2.

Bacillus sphaericus

Applications:

• Generation of square arrays of patterned metal, metal oxide, semiconductor, or other inorganic nanoclusters
• Patterning of biological macromolecules such as enzymes, protein receptors, or nucleic acids
• Physical and thermal stabilization of lipid-based structures, such as membrane monolayers, bilayers, and vesicles

Reassembly Conditions:

1. Dilute B. sphaericus S-Layer protein to ~20 µg/ml in high purity H₂O
2. Add CaCl₂ to a final concentration of 10 mM
3. Allow S-Layer to reassemble at interface for 1-16 hours
4. Wash substrate with buffer or high purity H₂O to remove unassembled S-Layer

User may need to optimize protein concentration, CaCl₂ concentration and incubation time depending on final applications

References:

Pum D. and Sleytr, U.B. (1995) Anisotropic Crystal Growth of the S-layer of Bacillus sphaericus CCM 2177 at the Air/Water Interface. Colloids and Surfaces 102: 99-104.

Gyorvary, E.S., Stein, O., Pum, D. and Sleytr, U.B. (2003) Self-assembly and Recrystallization of Bacterial S-layer Proteins at Silicon Supporots Imaged in Real Time by Atomic Force Microscopy. J. Microscopy. 212: 300- 306.

Specifications:
S-Layer: Square
Lattice Spacing: 12.5 nm
Pore size: 2.6 nm

Characteristics:

• S-layer protein is not cross-linked; it can be disassembled and reassembled.
• Works well on glass, silicon, and will readily reassemble on lipid membranes.

Sporosarcina ureae

Details:

Brightfield micrograph of
S. ureae cells

Sporosarcina ureae is a Gram-positive, motile, spore-forming coccus bacterium. This soil bacterium produces the enzyme urease, which breaks down urea to provide the organism with a source of nitrogen. Thus, it is thought to play a major role in urea decomposition in soil.

S. ureae produces an S-layer having a square pattern, which can be disassembled into monomers using chaotropic agents such as guanidine HCl. These monomers will reassemble upon addition of 10 mM CaCl_2.

Sporosarcina ureae

Applications:

• Generation of square arrays of patterned metal, metal oxide, semiconductor, or other inorganic nanoclusters
• Patterning of biological macromolecules such as enzymes, protein receptors, or nucleic acids
• Physical and thermal stabilization of lipid-based structures, such as membrane monolayers, bilayers, and vesicles

Reassembly Conditions:

1. Dilute S. ureae S-Layer protein to ~20 µg/ml in high purity H₂O
2. Add CaCl₂ to a final concentration of 10 mM
3. Allow S-Layer to reassemble at interface for 1-16 hours
4. Wash substrate with buffer or high purity H₂O to remove unassembled S-Layer

User may need to optimize protein concentration, CaCl₂ concentration and incubation time depending on final applications

References:

Engelhardt, H., Saxton, W.O. and Baumeister, W. (1986) Three-dimensional Structure of the Tetragonal Surface Layer of Sporosarcina ureae. J. Bacteriol. 168: 309-317.

Stewart, M. and Beveridge, T.J. (1980) Structure of the Regular Surface Layer of Sporosarcina ureae. J. Bacteriol. 142: 302-309.

Specifications:

S-Layer:
Oblique Parallelogram

Lattice Spacing:
11.6 x 8.6 nm

Pore size:
5.1 x 1.7 nm P.D.

Characteristics:

• S-layer protein is not cross-linked; it can be disassembled and reassembled.
• Works well on glass and silicon

Bacillus stearothermophilus

Details:

Brightfield micrograph of
B. stearothermophilus

Bacillus stearothermophilus is a Gram-positive spore-forming bacterium, with an optimal growth temperature of 55°C. B. stearothermophilus produces multiple S-layers depending on physiological conditions. The protein cataloged here is the oblique S-layers produced when the organism is cultured under aerobic conditions.

This S-layer can be disassembled into monomers using chaotropic agents such as guanidine HCl. These monomers will reassemble by the addition of 10 mM CaCl_2.

T.E.M. Images of Bacillus stearothermophilus.

B. stearothermophilus

Applications:

• Generation of oblique arrays of patterned metal, metal oxide, semiconductor, or other inorganic nanoclusters
• Patterning of biological macromolecules such as enzymes, protein receptors, or nucleic acids
• Physical and thermal stabilization of lipid-based structures, such as membrane monolayers, bilayers, and vesicles

Reassembly Conditions:

B. stearothermophilus S-Layers are cross-linked and can not be disassembled or reassembled.

1.  Dilute B. stearothermophilus S-Layer protein to ~20 µg/ml in high purity H₂O

2.  Add CaCl₂ to a final concentration of 10 mM

3.  Allow S-Layer to reassemble at interface for 1-16 hours

4.  Wash substrate with buffer or high purity H₂O to remove unassembled S-Layer

User may need to optimize protein concentration, CaCl₂ concentration, and incubation time depending on final applications

References:

Gruber, K. and Sleytr, U.B. (1988) Localized Insertion of New S-layer During Growth of Bacillus stearothermophilus Strains. Arch. Microbiol. 149: 485-491.

Mader, C., Huber, C., Moll, D., Sleytr, U.B. and Sára, M. (2004) Interaction of the Crystalline Bacterial Cell Surface Layer Protein SbsB and the Secondary Cell Wall Polymer of Geobacillus stearothermophilus PV72 Assessed by Real-Time Surface Plasmon Resonance Biosensor Technology. J. Bacteriol. 186: 1758-1768.

Specifications:

• S-layer protein is not cross-linked; it can be disassembled and reassembled.
• Works well on glass and silicon

Lactobacillus helveticus

Details:

Brightfield micrograph of
L. helveticus cells

Lactobacillus helveticus is a Gram-positive, nonspore-forming lactic acid bacterium. It is used extensively in cheese making, serving as a culture ingredient for Swiss, Parmesan, Romano, Provolone and Mozzarella cheeses.

S-layers of the organism form an oblique pattern that can be disassembled using lithium chloride, and can reassemble on various surfaces.

T.E.M. Images of Lactobacillus helveticus.

Lactobacillus helveticus

Applications:

• Generation of oblique arrays of patterned metal, metal oxide, semiconductor, or other inorganic nanoclusters
• Patterning of biological macromolecules such as enzymes, protein receptors, or nucleic acids
• Physical and thermal stabilization of lipid-based structures, such as membrane monolayers, bilayers, and vesicles

Reassembly Conditions:

1. Dilute L. helveticus S-Layer protein to ~20 µg/ml (in 5 M LiCl) 1:10 in high purity H₂O
2. Immediately aliquot S-Layer protein on substrate for reassembly
3. Allow S-Layer to reassemble at interface for 1-16 hours
4. Wash substrate with buffer or high purity H₂O to remove unassembled S-Layer

User may need to optimize protein concentration and incubation time depending on final applications

References:

Lortal, S., van Heijenoort, J., Gruber, K. and Sleytr, U.B. (1992) S-layer of Lactobacillus helveticus ATCC 12046: Isolation, Chemical Characterization and Re-formation After Extraction with Lithium Chloride. J. Gen. Micro. 138: 611-618.

Schar-Zammaretti, P. and Ubbink, J. (2003) The Cell Wall of Lactic Acid Bacteria: Surface Constituents and Macromolecular Conformations. Biophys. J. 85: 4076-4092.