Difference between revisions of "Part:BBa K4863005"

Revision as of 05:05, 11 October 2023

T7_hpCA_Slp

alpha type carbonic anhydrase fused with s-layer protein via a flexible linker for surface display of hpCA on the cell surface of Synechocystis PCC6803. This yields a cell completely covered in the fusion protein. Surface display maximizes hpCA's binding to substrate and ensures its stability.

Usage and Biology

The fusion of a protein to the Slp N-terminus has proved to be an effective system for the functional surface display of cyanobacteria. For the project of this year, we displayed α-carbonic anhydrase hpCA on the Synechocystis PCC6803 cell surface with the fusion with Slp on the cell surface for the production of calcium carbonate precipitates.

hpCA is a metalloenzymes responsible for catalyzing the interconversion of carbon dioxide (CO2) to bicarbonate ions (HCO3−). It is an α-carbonic anhydrase isolated from the bacterium Helicobacter pylori 26695, codon optimized for expression in E. Coli BL21(DE3). Slp forms the paracrystalline S-layer by forming a regular lattice through an entropy-driven process. It has an undefined signal peptide at the C-terminus.

Sequence and Features

Reference

Bury-Moné, S., Mendz, G. L., Ball, G. E., Thibonnier, M., Stingl, K., Ecobichon, C., Avé, P., Huerre, M., Labigne, A., Thiberge, J.-M., & DeReuse, H. (2008). Roles of α and β carbonic anhydrases of Helicobacter pylori in the urease-dependent response to acidity and in colonization of the murine gastric mucosa. Infection and Immunity, 76(2), 497-509. https://doi.org/10.1128/iai.00993-07

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.

Chirica, L. C., Petersson, C., Hurtig, M., Jonsson, B.-H., Borén, T., & Lindskog, S. (2002). Expression and localization of α- and β-carbonic anhydrase in Helicobacter pylori. Biochimica Et Biophysica Acta (BBA) - Proteins and Proteomics, 1601(2), 192-199. https://doi.org/10.1016/s1570-9639(02)00467-3

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.

Zhu, Y., Liu, Y., Ai, M., & Jia, X. (2022). Surface display of carbonic anhydrase on Escherichia coli for CO2 capture and mineralization. Synthetic and Systems Biotechnology, 7(1), 460-473. https://doi.org/10.1016/j.synbio.2021.11.008