Lactococcin B Is Inactivated by Intrinsic Proteinase PrtP Digestion in Lactococcus lactis subsp. lactis BGMN1-501
Autori
Vukotić, GoranPolović, Natalija
Mirković, Nemanja
Jovčić, Branko
Stanisavljević, Nemanja
Fira, Đorđe
Kojić, Milan
Članak u časopisu (Objavljena verzija)
Metapodaci
Prikaz svih podataka o dokumentuApstrakt
In our previous study we demonstrated that proteinase PrtP is able to impair bacteriocin LcnB activity, despite being produced by the same organism and encoded by the same plasmid. However, precise mechanism of this action, i.e., the exact cleavage site within LcnB bacteriocin, as well as its effect on antimicrobial activity of the resulting peptide remained vague. Here we further explored the interplay between these two proteins and defined, using mass spectrometry, that this unusual hydrolysis indeed occurs in vivo, between the sixth and seventh amino acid on the N terminus of LcnB. To address whether the cleaved form of LcnB retains any level of activity, both recombinant and chemically synthesized variant of truncated LcnB were engineered and produced, but demonstrated no antimicrobial activity. When LcnB was recombinantly overexpressed and subjected to PrtP digestion, the change in its antimicrobial activity was monitored and the degradation products analyzed with reverse-phase hi...gh-pressure liquid chromatography. The results confirmed the inactivity of the truncated LcnB and additionally corroborated the PrtP cleavage site in LcnB bacteriocin. In addition, it was demonstrated that, once truncated, LcnB is not able to bind its receptor and is susceptible to additional hydrolysis. This is the first report on proteolytic inactivation of bacteriocins inside the same bacterial host.
Ključne reči:
proteinase PrtP / Lactococcus lactis / inactivation / hydrolysis / bacteriocin LcnBIzvor:
Frontiers in Microbiology, 2019, 10Izdavač:
- Frontiers Media Sa, Lausanne
Finansiranje / projekti:
- Izučavanje gena i molekularnih mehanizama u osnovi probiotičke aktivnosti bakterija mlečne kiseline izolovanih sa područja zapadnog Balkana (RS-MESTD-Basic Research (BR or ON)-173019)
- Molekularna karakterizacija bakterija iz rodova Bacillus i Pseudomonas kao potencijalnih agenasa za biološku kontrolu (RS-MESTD-Basic Research (BR or ON)-173026)
DOI: 10.3389/fmicb.2019.00874
ISSN: 1664-302X
WoS: 000465419600001
Scopus: 2-s2.0-85068164054
Institucija/grupa
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Vukotić, Goran AU - Polović, Natalija AU - Mirković, Nemanja AU - Jovčić, Branko AU - Stanisavljević, Nemanja AU - Fira, Đorđe AU - Kojić, Milan PY - 2019 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/1251 AB - In our previous study we demonstrated that proteinase PrtP is able to impair bacteriocin LcnB activity, despite being produced by the same organism and encoded by the same plasmid. However, precise mechanism of this action, i.e., the exact cleavage site within LcnB bacteriocin, as well as its effect on antimicrobial activity of the resulting peptide remained vague. Here we further explored the interplay between these two proteins and defined, using mass spectrometry, that this unusual hydrolysis indeed occurs in vivo, between the sixth and seventh amino acid on the N terminus of LcnB. To address whether the cleaved form of LcnB retains any level of activity, both recombinant and chemically synthesized variant of truncated LcnB were engineered and produced, but demonstrated no antimicrobial activity. When LcnB was recombinantly overexpressed and subjected to PrtP digestion, the change in its antimicrobial activity was monitored and the degradation products analyzed with reverse-phase high-pressure liquid chromatography. The results confirmed the inactivity of the truncated LcnB and additionally corroborated the PrtP cleavage site in LcnB bacteriocin. In addition, it was demonstrated that, once truncated, LcnB is not able to bind its receptor and is susceptible to additional hydrolysis. This is the first report on proteolytic inactivation of bacteriocins inside the same bacterial host. PB - Frontiers Media Sa, Lausanne T2 - Frontiers in Microbiology T1 - Lactococcin B Is Inactivated by Intrinsic Proteinase PrtP Digestion in Lactococcus lactis subsp. lactis BGMN1-501 VL - 10 DO - 10.3389/fmicb.2019.00874 ER -
@article{ author = "Vukotić, Goran and Polović, Natalija and Mirković, Nemanja and Jovčić, Branko and Stanisavljević, Nemanja and Fira, Đorđe and Kojić, Milan", year = "2019", abstract = "In our previous study we demonstrated that proteinase PrtP is able to impair bacteriocin LcnB activity, despite being produced by the same organism and encoded by the same plasmid. However, precise mechanism of this action, i.e., the exact cleavage site within LcnB bacteriocin, as well as its effect on antimicrobial activity of the resulting peptide remained vague. Here we further explored the interplay between these two proteins and defined, using mass spectrometry, that this unusual hydrolysis indeed occurs in vivo, between the sixth and seventh amino acid on the N terminus of LcnB. To address whether the cleaved form of LcnB retains any level of activity, both recombinant and chemically synthesized variant of truncated LcnB were engineered and produced, but demonstrated no antimicrobial activity. When LcnB was recombinantly overexpressed and subjected to PrtP digestion, the change in its antimicrobial activity was monitored and the degradation products analyzed with reverse-phase high-pressure liquid chromatography. The results confirmed the inactivity of the truncated LcnB and additionally corroborated the PrtP cleavage site in LcnB bacteriocin. In addition, it was demonstrated that, once truncated, LcnB is not able to bind its receptor and is susceptible to additional hydrolysis. This is the first report on proteolytic inactivation of bacteriocins inside the same bacterial host.", publisher = "Frontiers Media Sa, Lausanne", journal = "Frontiers in Microbiology", title = "Lactococcin B Is Inactivated by Intrinsic Proteinase PrtP Digestion in Lactococcus lactis subsp. lactis BGMN1-501", volume = "10", doi = "10.3389/fmicb.2019.00874" }
Vukotić, G., Polović, N., Mirković, N., Jovčić, B., Stanisavljević, N., Fira, Đ.,& Kojić, M.. (2019). Lactococcin B Is Inactivated by Intrinsic Proteinase PrtP Digestion in Lactococcus lactis subsp. lactis BGMN1-501. in Frontiers in Microbiology Frontiers Media Sa, Lausanne., 10. https://doi.org/10.3389/fmicb.2019.00874
Vukotić G, Polović N, Mirković N, Jovčić B, Stanisavljević N, Fira Đ, Kojić M. Lactococcin B Is Inactivated by Intrinsic Proteinase PrtP Digestion in Lactococcus lactis subsp. lactis BGMN1-501. in Frontiers in Microbiology. 2019;10. doi:10.3389/fmicb.2019.00874 .
Vukotić, Goran, Polović, Natalija, Mirković, Nemanja, Jovčić, Branko, Stanisavljević, Nemanja, Fira, Đorđe, Kojić, Milan, "Lactococcin B Is Inactivated by Intrinsic Proteinase PrtP Digestion in Lactococcus lactis subsp. lactis BGMN1-501" in Frontiers in Microbiology, 10 (2019), https://doi.org/10.3389/fmicb.2019.00874 . .