High yield expression of leptospirosis vaccine candidates LigA and LipL32 in the methylotrophic yeast Pichia pastoris.
BACKGROUND
Leptospirosis, a zoonosis caused by Leptospira spp., is recognized as an emergent infectious disease. Due to the lack of adequate diagnostic tools, vaccines are an attractive intervention strategy. Recombinant proteins produced in Escherichia coli have demonstrated promising results, albeit with variable efficacy. Pichia pastoris is an alternative host with several advantages for the production of recombinant proteins.


RESULTS
The vaccine candidates LigANI and LipL32 were cloned and expressed in P. pastoris as secreted proteins. Large-scale expression resulted in a yield of 276 mg/L for LigANI and 285 mg/L for LipL32. The recombinant proteins were glycosylated and were recognized by antibodies present in the sera of patients with severe leptospirosis.


CONCLUSIONS
The expression of LigANI and LipL32 in P. pastoris resulted in a significant increase in yield compared to expression in E. coli. In addition, the proteins were secreted, allowing for easy purification, and retained the antigenic characteristics of the native proteins, demonstrating their potential application as subunit vaccine candidates.
Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
Daiane D
DD
Hartwig
Thaís L
TL
Oliveira
Fabiana K
FK
Seixas
Karine M
KM
Forster
Caroline
C
Rizzi
Cláudia P
CP
Hartleben
Alan J A
AJ
McBride
Odir A
OA
Dellagostin
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D001428:Bacterial Vaccines / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007919:Leptospira / Q000201:enzymology / Q000276:immunology*; D007922:Leptospirosis / Q000276:immunology*; D008074:Lipoproteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D022223:Vaccines, Subunit / Q000235:genetics / Q000276:immunology / Q000378:metabolism
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; C406589:LipL32 protein, Leptospira; D008074:Lipoproteins; D011994:Recombinant Proteins; D022223:Vaccines, Subunit