Screening for New Surface Anchoring Domains for Lactococcus lactis
The display of recombinant proteins on bacterial surfaces is a developing research area with a wide range of potential biotechnological applications. The lactic acid bacterium Lactococcus lactis is an attractive host for such surface display, and a promising vector for in vivo delivery of bioactive...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2019-08-01
|
| Series: | Frontiers in Microbiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2019.01879/full |
| _version_ | 1811235672757895168 |
|---|---|
| author | Tina Vida Plavec Tina Vida Plavec Borut Štrukelj Borut Štrukelj Aleš Berlec Aleš Berlec |
| author_facet | Tina Vida Plavec Tina Vida Plavec Borut Štrukelj Borut Štrukelj Aleš Berlec Aleš Berlec |
| author_sort | Tina Vida Plavec |
| collection | DOAJ |
| description | The display of recombinant proteins on bacterial surfaces is a developing research area with a wide range of potential biotechnological applications. The lactic acid bacterium Lactococcus lactis is an attractive host for such surface display, and a promising vector for in vivo delivery of bioactive proteins. Surface-displayed recombinant proteins are usually anchored to the bacterial cell wall through anchoring domains. Here, we investigated alternatives to the commonly applied lactococcal lysine motif (LysM)-containing surface anchoring domain, the C-terminus of AcmA (cAcmA). We screened 15 anchoring domains of lactococcal or phage origins that belong to the Pfam categories LPXTG, LysM, CW_1, Cpl-7, WxL, SH3, and ChW, which can provide non-covalent or covalent binding to the cell wall. LPXTG, LysM, the duplicated CW_1 and SH3 domains promoted significant surface display of two model proteins, B domain and DARPin I07, although the display achieved was lower than that for the reference anchoring domain, cAcmA. On the other hand, the ChW-containing anchoring domain of the lactococcal phage AM12 endolysin (cAM12) demonstrated surface display comparable to that of cAcmA. The anchoring ability of cAM12 was confirmed by enabling non-covalent heterologous anchoring of the B domain on wild-type bacteria, as well as anchoring of CXCL8-binding evasin-3, which provided potential therapeutic applicability; both were displayed to an extent comparable to that of cAcmA. We have thereby demonstrated the effective use of different protein anchoring domains in L. lactis, with ChW-containing cAM12 the most promising alternative to the established approaches for surface display on L. lactis. |
| first_indexed | 2024-04-12T11:56:01Z |
| format | Article |
| id | doaj.art-828ae09729fe4dafa3777c4ea77acea3 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-04-12T11:56:01Z |
| publishDate | 2019-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-828ae09729fe4dafa3777c4ea77acea32022-12-22T03:34:01ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2019-08-011010.3389/fmicb.2019.01879466935Screening for New Surface Anchoring Domains for Lactococcus lactisTina Vida Plavec0Tina Vida Plavec1Borut Štrukelj2Borut Štrukelj3Aleš Berlec4Aleš Berlec5Department of Biotechnology, Jožef Stefan Institute, Ljubljana, SloveniaFaculty of Pharmacy, University of Ljubljana, Ljubljana, SloveniaDepartment of Biotechnology, Jožef Stefan Institute, Ljubljana, SloveniaFaculty of Pharmacy, University of Ljubljana, Ljubljana, SloveniaDepartment of Biotechnology, Jožef Stefan Institute, Ljubljana, SloveniaFaculty of Pharmacy, University of Ljubljana, Ljubljana, SloveniaThe display of recombinant proteins on bacterial surfaces is a developing research area with a wide range of potential biotechnological applications. The lactic acid bacterium Lactococcus lactis is an attractive host for such surface display, and a promising vector for in vivo delivery of bioactive proteins. Surface-displayed recombinant proteins are usually anchored to the bacterial cell wall through anchoring domains. Here, we investigated alternatives to the commonly applied lactococcal lysine motif (LysM)-containing surface anchoring domain, the C-terminus of AcmA (cAcmA). We screened 15 anchoring domains of lactococcal or phage origins that belong to the Pfam categories LPXTG, LysM, CW_1, Cpl-7, WxL, SH3, and ChW, which can provide non-covalent or covalent binding to the cell wall. LPXTG, LysM, the duplicated CW_1 and SH3 domains promoted significant surface display of two model proteins, B domain and DARPin I07, although the display achieved was lower than that for the reference anchoring domain, cAcmA. On the other hand, the ChW-containing anchoring domain of the lactococcal phage AM12 endolysin (cAM12) demonstrated surface display comparable to that of cAcmA. The anchoring ability of cAM12 was confirmed by enabling non-covalent heterologous anchoring of the B domain on wild-type bacteria, as well as anchoring of CXCL8-binding evasin-3, which provided potential therapeutic applicability; both were displayed to an extent comparable to that of cAcmA. We have thereby demonstrated the effective use of different protein anchoring domains in L. lactis, with ChW-containing cAM12 the most promising alternative to the established approaches for surface display on L. lactis.https://www.frontiersin.org/article/10.3389/fmicb.2019.01879/fullsurface displayLactococcus lactisanchorphage AM12ChWLPXTG |
| spellingShingle | Tina Vida Plavec Tina Vida Plavec Borut Štrukelj Borut Štrukelj Aleš Berlec Aleš Berlec Screening for New Surface Anchoring Domains for Lactococcus lactis Frontiers in Microbiology surface display Lactococcus lactis anchor phage AM12 ChW LPXTG |
| title | Screening for New Surface Anchoring Domains for Lactococcus lactis |
| title_full | Screening for New Surface Anchoring Domains for Lactococcus lactis |
| title_fullStr | Screening for New Surface Anchoring Domains for Lactococcus lactis |
| title_full_unstemmed | Screening for New Surface Anchoring Domains for Lactococcus lactis |
| title_short | Screening for New Surface Anchoring Domains for Lactococcus lactis |
| title_sort | screening for new surface anchoring domains for lactococcus lactis |
| topic | surface display Lactococcus lactis anchor phage AM12 ChW LPXTG |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2019.01879/full |
| work_keys_str_mv | AT tinavidaplavec screeningfornewsurfaceanchoringdomainsforlactococcuslactis AT tinavidaplavec screeningfornewsurfaceanchoringdomainsforlactococcuslactis AT borutstrukelj screeningfornewsurfaceanchoringdomainsforlactococcuslactis AT borutstrukelj screeningfornewsurfaceanchoringdomainsforlactococcuslactis AT alesberlec screeningfornewsurfaceanchoringdomainsforlactococcuslactis AT alesberlec screeningfornewsurfaceanchoringdomainsforlactococcuslactis |