High level expression and glycosylation of recombinant Mycobacterium tuberculosis Ala-Pro-rich antigen in Pichia pastoris.
Protein Expr Purif
; 150: 67-71, 2018 10.
Article
em En
| MEDLINE
| ID: mdl-29753122
ABSTRACT
The Ala-Pro-rich Antigen (Apa) from Mycobacterium tuberculosis is a mannosylated protein with immunogenic and antigenic properties. The O-mannosylation is essential for its biological function in the process of colonization and invasion of host cells by M. tuberculosis. In this work, the gene encoding Apa was cloned from M. tuberculosis and expressed in Pichia pastoris GS115. In shake-flasks, the recombinant Apa was secreted into the culture media and purified with a yield of 0.6â¯g/L. Both N- and O-glycans were found in recombinant Apa. In P. pastoris the known M. tuberculosis-derived O-glycosites of Apa were modified with short chains of mannose units, and a few additional glycosylation sites were also observed. Therefore, the recombinant Apa expressed in P. pastoris has similar but not identical O-mannose patterns to the native protein from M. tuberculosis. P. pastoris and mycobacteria share similarities in the protein O-glycosylation pathway. Thus P. pastoris could be a potential powerful expression system to produce mycobacteria-derived glycoproteins.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Pichia
/
Expressão Gênica
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Mycobacterium tuberculosis
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Antígenos de Bactérias
Idioma:
En
Revista:
Protein Expr Purif
Assunto da revista:
BIOLOGIA MOLECULAR
Ano de publicação:
2018
Tipo de documento:
Article
País de afiliação:
China