| RRC ID |
66788
|
| 著者 |
Gordillo TB, Palumbo MC, Allievi MC, Fernández Do Porto DA, Ruzal SM, Palomino MM.
|
| タイトル |
Strategies to display heterologous proteins on the cell surface of lactic acid bacteria using as anchor the C-terminal domain of Lactobacillus acidophilus SlpA.
|
| ジャーナル |
World J Microbiol Biotechnol
|
| Abstract |
The surface-layer (S-layer) protein of Lactobacillus acidophilus is a crystalline array of self-assembling subunits, non-covalently bound to the most outer cell wall envelope, which constitutes up to 20% of the total cell protein content. These attributes make S-layer proteins an excellent anchor for the development of microbial cell-surface display systems. In L. acidophilus, the S-layer is formed predominantly by the protein SlpA. We have previously shown that the C-terminal domain of SlpA is responsible for the cell wall anchorage on L. acidophilus ATCC 4356. In the present study, we evaluated the C-terminal domain of SlpA of L. acidophilus ATCC 4356 as a potential anchor domain to display functional proteins on the surface of non-genetically modified lactic acid bacteria (LAB). To this end, green fluorescent protein (GFP)-CTSlpA was firstly produced in Escherichia coli and the recombinant proteins were able to spontaneously bind to the cell wall of LAB in a binding assay. GFP was successfully displayed on the S-layer stripped surface of L. acidophilus. Both the binding stability and cell survival of L. acidophilus decorated with the recombinant protein were then studied in simulated gastrointestinal conditions. Furthermore, NaCl was tested as a safer alternative to LiCl for S-layer removal. This study presents the development of a protein delivery platform involving L. acidophilus, a microorganism generally regarded as safe, which utilizes the contiguous, non-covalently attached S-layer at the cell surface of the bacterium without introducing any genetic modification.
|
| 巻・号 |
36(11)
|
| ページ |
169
|
| 公開日 |
2020-10-12
|
| DOI |
10.1007/s11274-020-02945-9
|
| PII |
10.1007/s11274-020-02945-9
|
| PMID |
33043388
|
| MeSH |
Bacterial Proteins / genetics
Bacterial Proteins / metabolism
Cell Membrane / chemistry*
Cell Wall / chemistry
Cloning, Molecular
Culture Media / chemistry
Escherichia coli / genetics
Escherichia coli / metabolism
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism
Lactobacillales / metabolism*
Lactobacillus acidophilus / metabolism*
Membrane Glycoproteins / genetics
Membrane Glycoproteins / metabolism
Membrane Proteins / metabolism
Microbial Viability
Microscopy, Electron, Transmission
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / metabolism
|
| リソース情報 |
| 一般微生物 |
JCM5818 |
