Difference between revisions of "Part:BBa K4863002"
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<partinfo>BBa_K4863002 short</partinfo> | <partinfo>BBa_K4863002 short</partinfo> | ||
| − | S-layer protein (Slp) is a native protein on the cell surface of cyanobacteria. It is encoded by the gene sll1951 in <i>Synechocystis</i> PCC6803. This protein forms a lattice that is about 30nm wide that covers the cell surface, forming the paracrystalline S-layer. It has an undefined signal peptide at C-terminus. | + | S-layer protein (Slp) is a native protein on the cell surface of cyanobacteria. It is encoded by the gene sll1951 in <i>Synechocystis</i> PCC6803. This protein forms a lattice that is about 30nm wide that covers the cell surface, forming the paracrystalline S-layer. It has an undefined signal peptide at C-terminus (Trautner and Vermaas, 2013). |
Fusion of a protein to the N-terminus of Slp is shown to be an effective system for functional surface display since it would yield a cell entirely covered with the fusion protein on the outermost layer. This year, we display the α-carbonic anhydrase hpCA on the cell surface of <i>Synechocystis</i> PCC6803 via fusion with Slp. We also display the 13 amino acid-long peptide chain SpyTag(<partinfo>K1159200</partinfo>) on the cell surface of <i>Synechocystis</i> PCC6803 via fusion with Slp, thus achieving surface display of a hpCA-SpyCatcher complex (<partinfo>K4863000</partinfo> and <partinfo>K1159201</partinfo>) via covalent bonding between the displayed SpyTag and the SpyCatcher. | Fusion of a protein to the N-terminus of Slp is shown to be an effective system for functional surface display since it would yield a cell entirely covered with the fusion protein on the outermost layer. This year, we display the α-carbonic anhydrase hpCA on the cell surface of <i>Synechocystis</i> PCC6803 via fusion with Slp. We also display the 13 amino acid-long peptide chain SpyTag(<partinfo>K1159200</partinfo>) on the cell surface of <i>Synechocystis</i> PCC6803 via fusion with Slp, thus achieving surface display of a hpCA-SpyCatcher complex (<partinfo>K4863000</partinfo> and <partinfo>K1159201</partinfo>) via covalent bonding between the displayed SpyTag and the SpyCatcher. | ||
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Following the potential of cyanobacteria as a cell factory for biofuel or biomaterials, functional surface display of proteins in cyanobacteria is beneficial in many ways: it can allow for better enzyme performance via improved binding with substrate, achieve immobilization of cells, and more. Cyanobacteria have a thick peptidoglycan layer and an additional S-layer, making effective surface display difficult. S-layer protein (Slp) is the product of sll1951 in <i>Synechocystis</i>. It makes up the extracellular paracrystalline S-layer outside of the cyanobacterial cell wall and is the only component of this outermost layer. Slp has an undefined signal peptide at C-terminus, and fusion of another protein at the N-terminus of Slp would yield a <i>Synechocystis</i> cell completely covered with the fusion protein on the outermost layer and is thus an effective method for surface display (Cengic et al., 2018). | Following the potential of cyanobacteria as a cell factory for biofuel or biomaterials, functional surface display of proteins in cyanobacteria is beneficial in many ways: it can allow for better enzyme performance via improved binding with substrate, achieve immobilization of cells, and more. Cyanobacteria have a thick peptidoglycan layer and an additional S-layer, making effective surface display difficult. S-layer protein (Slp) is the product of sll1951 in <i>Synechocystis</i>. It makes up the extracellular paracrystalline S-layer outside of the cyanobacterial cell wall and is the only component of this outermost layer. Slp has an undefined signal peptide at C-terminus, and fusion of another protein at the N-terminus of Slp would yield a <i>Synechocystis</i> cell completely covered with the fusion protein on the outermost layer and is thus an effective method for surface display (Cengic et al., 2018). | ||
| − | The S-layer of cyanobacteria, which entirely constitutes of the Slp protein, is believed to be involved in carbon sequestration and other physiochemical processes. Slp self-assembles into lattice form via an entropy-driven process in extracellular space. In <i>Synechocystis</i> PCC 6803, the sll1951 gene, which is a 5226bp open reading frame, encodes for the hemolysin-like S-layer protein. | + | The S-layer of cyanobacteria, which entirely constitutes of the Slp protein, is believed to be involved in carbon sequestration and other physiochemical processes. Slp self-assembles into lattice form via an entropy-driven process in extracellular space(Šmarda et al., 2002). In <i>Synechocystis</i> PCC 6803, the sll1951 gene, which is a 5226bp open reading frame, encodes for the hemolysin-like S-layer protein. |
===Characterization=== | ===Characterization=== | ||
Revision as of 13:20, 9 October 2023
S-layer protein (Slp)
S-layer protein (Slp) is a native protein on the cell surface of cyanobacteria. It is encoded by the gene sll1951 in Synechocystis PCC6803. This protein forms a lattice that is about 30nm wide that covers the cell surface, forming the paracrystalline S-layer. It has an undefined signal peptide at C-terminus (Trautner and Vermaas, 2013).
Fusion of a protein to the N-terminus of Slp is shown to be an effective system for functional surface display since it would yield a cell entirely covered with the fusion protein on the outermost layer. This year, we display the α-carbonic anhydrase hpCA on the cell surface of Synechocystis PCC6803 via fusion with Slp. We also display the 13 amino acid-long peptide chain SpyTag(BBa_K1159200) on the cell surface of Synechocystis PCC6803 via fusion with Slp, thus achieving surface display of a hpCA-SpyCatcher complex (BBa_K4863000 and BBa_K1159201) via covalent bonding between the displayed SpyTag and the SpyCatcher.
Usage and Biology
Following the potential of cyanobacteria as a cell factory for biofuel or biomaterials, functional surface display of proteins in cyanobacteria is beneficial in many ways: it can allow for better enzyme performance via improved binding with substrate, achieve immobilization of cells, and more. Cyanobacteria have a thick peptidoglycan layer and an additional S-layer, making effective surface display difficult. S-layer protein (Slp) is the product of sll1951 in Synechocystis. It makes up the extracellular paracrystalline S-layer outside of the cyanobacterial cell wall and is the only component of this outermost layer. Slp has an undefined signal peptide at C-terminus, and fusion of another protein at the N-terminus of Slp would yield a Synechocystis cell completely covered with the fusion protein on the outermost layer and is thus an effective method for surface display (Cengic et al., 2018).
The S-layer of cyanobacteria, which entirely constitutes of the Slp protein, is believed to be involved in carbon sequestration and other physiochemical processes. Slp self-assembles into lattice form via an entropy-driven process in extracellular space(Šmarda et al., 2002). In Synechocystis PCC 6803, the sll1951 gene, which is a 5226bp open reading frame, encodes for the hemolysin-like S-layer protein.
Characterization
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 3112
Illegal EcoRI site found at 5092
Illegal XbaI site found at 318
Illegal PstI site found at 434
Illegal PstI site found at 4034 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 3112
Illegal EcoRI site found at 5092
Illegal NheI site found at 741
Illegal NheI site found at 838
Illegal PstI site found at 434
Illegal PstI site found at 4034 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 3112
Illegal EcoRI site found at 5092
Illegal BglII site found at 2253
Illegal BamHI site found at 3159
Illegal BamHI site found at 4473
Illegal XhoI site found at 376 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 3112
Illegal EcoRI site found at 5092
Illegal XbaI site found at 318
Illegal PstI site found at 434
Illegal PstI site found at 4034 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 3112
Illegal EcoRI site found at 5092
Illegal XbaI site found at 318
Illegal PstI site found at 434
Illegal PstI site found at 4034
Illegal AgeI site found at 2140
Illegal AgeI site found at 2323
Illegal AgeI site found at 2671
Illegal AgeI site found at 2731
Illegal AgeI site found at 4252 - 1000COMPATIBLE WITH RFC[1000]
Reference
Cengic I, Uhlén M, Hudson EP. (2018). Surface Display of Small Affinity Proteins on Synechocystis sp. Strain PCC 6803 Mediated by Fusion to the Major Type IV Pilin PilA1. Journal of Bacteriology, 200(16). https://doi.org/10.1128/JB.00270-18.
Šmarda, J., Šmajs, D., Komrska, J., & Krzyžánek, V. (2002). S-layers on cell walls of cyanobacteria. Micron, 33(3), 257-277. https://doi.org/10.1016/s0968-4328(01)00031-2.
Trautner, C., & Vermaas, W. F. J. (2013). The sll1951 gene encodes the surface layer protein of synechocystis sp. strain PCC 6803. Journal of Bacteriology, 195(23), 5370-5380. https://doi.org/10.1128/jb.00615-13.