High-yield production of a functional bacteriophage lysin with antipneumococcal activity using a plant virus-based expression system.
Streptococcus pneumoniae is the causative agent of several serious infectious diseases. It is becoming increasingly antibiotic resistant worldwide, and thus new antimicrobials are needed. One alternative to antibiotics may be the use of peptidoglycan hydrolases, the bacteriophage lytic enzymes. In this study, we demonstrated high level expression of the S. pneumoniae bacteriophage lysin Pal in Nicotiana benthamiana - TMV (Tobacco Mosaic Virus) transient expression system. The protein was purified to homogeneity and tested for streptococci killing activity in vitro and in vivo. In vitro, Pal was able to lyse three tested S. pneumoniae strains: NCTC12695, NCTC12977 and NCTC11888. The treatment of BALB/c mice with 100 μg, 200 μg and 400 μg of Pal 1h post-challenge with double lethal dose of S. pneumoniae NCTC12695 strain showed a clear dose response and protected from lethal sepsis 30%, 40% and 50% of mice, respectively. The improved mice survival correlated with decreased blood bacterial titers. In conclusion, these results suggest that plant-expressed bacteriophage lysins may have potential use as antimicrobial agents.
Nomads UAB, Graičiūno 8, LT-02241 Vilnius, Lithuania; Institute of Biotechnology of Vilnius University, Graičiūno 8, LT-02241 Vilnius, Lithuania.
Urtė
U
Starkevič
Nomad Bioscience GmbH, Biozentrum Halle, Weinbergweg 22, D-06120 Halle (Saale), Germany.
Luisa
L
Bortesi
LUHS Veterinary Academy, Tilžės 18, LT-47181 Kaunas, Lithuania.
Marius
M
Virgailis
LUHS Veterinary Academy, Tilžės 18, LT-47181 Kaunas, Lithuania.
Modestas
M
Ružauskas
Nomad Bioscience GmbH, Biozentrum Halle, Weinbergweg 22, D-06120 Halle (Saale), Germany.
Anatoli
A
Giritch
Nomads UAB, Graičiūno 8, LT-02241 Vilnius, Lithuania. Electronic address: ausra@nomadsbio.lt.
Aušra
A
Ražanskienė
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology / Q000627:therapeutic use; D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism* / Q000494:pharmacology / Q000627:therapeutic use; D016470:Bacteremia / Q000188:drug therapy; D008807:Mice, Inbred BALB C; D009088:Mucoproteins; D018515:Plant Leaves / Q000502:physiology / Q000821:virology; D017108:Streptococcus Phages; D013296:Streptococcus pneumoniae / Q000187:drug effects / Q000502:physiology; D014026:Nicotiana / Q000502:physiology / Q000821:virology*; D014027:Tobacco Mosaic Virus / Q000502:physiology*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology / Q000627:therapeutic use
D000900:Anti-Bacterial Agents; D009088:Mucoproteins; D014764:Viral Proteins; C010393:lysin, gastropoda; D000581:Amidohydrolases; C493909:Pal protein, pneumococcal bacteriophage Dp-1
Bacteriophage lysin; Mice bacteremia and survival; Plant virus-based transient expression; Streptococcus pneumoniae