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http://ru.facmed.unam.mx/jspui/handle/FACMED_UNAM/A102
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Campo DC | Valor | Lengua/Idioma |
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dc.contributor.author | Bando Campos, Carlos Giroshi | |
dc.contributor.author | Juarez Lopez, Daniel | |
dc.contributor.author | Roman Gonzalez, Sergio Agustin | |
dc.contributor.author | Castillo Rodal, Antonia Isabel | |
dc.contributor.author | Olvera Carranza, Clarita | |
dc.contributor.author | Lopez Vidal, Yolanda | |
dc.contributor.author | Arreguin Espinosa de los Monteros, Roberto | |
dc.contributor.author | Espitia Pinzon, Clara Ines | |
dc.contributor.author | Trujillo Roldan, Mauricio Alberto | |
dc.contributor.author | Valdez Cruz, Norma Adriana | |
dc.coverage.spatial | GB | |
dc.date.accessioned | 2019-06-17T17:23:06Z | - |
dc.date.available | 2019-06-17T17:23:06Z | - |
dc.date.issued | 2019 | |
dc.identifier.uri | http://ru.facmed.unam.mx/jspui/handle/FACMED_UNAM/A102 | - |
dc.description.abstract | BACKGROUND: Pichia pastoris (syn. Komagataella phaffii) is one of the most highly utilized eukaryotic expression systems for the production of heterologous glycoproteins, being able to perform both N- and O-mannosylation. In this study, we present the expression in P. pastoris of an O-mannosylated recombinant version of the 38 kDa glycolipoprotein PstS-1 from Mycobacterium tuberculosis (Mtb), that is similar in primary structure to the native secreted protein. RESULTS: The recombinant PstS-1 (rPstS-1) was produced without the native lipidation signal. Glycoprotein expression was under the control of the methanol-inducible promoter pAOX1, with secretion being directed by the ?-mating factor secretion signal. Production of rPstS-1 was carried out in baffled shake flasks (BSFs) and controlled bioreactors. A production up to ~ 46 mg/L of the recombinant protein was achieved in both the BSFs and the bioreactors. The recombinant protein was recovered from the supernatant and purified in three steps, achieving a preparation with 98% electrophoretic purity. The primary and secondary structures of the recombinant protein were characterized, as well as its O-mannosylation pattern. Furthermore, a cross-reactivity analysis using serum antibodies from patients with active tuberculosis demonstrated recognition of the recombinant glycoprotein, indirectly indicating the similarity between the recombinant PstS-1 and the native protein from Mtb. CONCLUSIONS: rPstS-1 (98.9% sequence identity, O-mannosylated, and without tags) was produced and secreted by P. pastoris, demonstrating that this yeast is a useful cell factory that could also be used to produce other glycosylated Mtb antigens. The rPstS-1 could be used as a tool for studying the role of this molecule during Mtb infection, and to develop and improve vaccines or kits based on the recombinant protein for serodiagnosis. | |
dc.language.iso | en | |
dc.publisher | BioMed Central | |
dc.rights | openAccess | |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0 | |
dc.subject | Bacteriología | |
dc.subject | Mycobacterium tuberculosis | |
dc.subject | Antígeno | |
dc.subject | Glicoproteína | |
dc.subject | Pichia pastoris | |
dc.subject | PstS-1 | |
dc.subject | Komagataella phaffii | |
dc.subject.classification | Medicina y Ciencias de la Salud | |
dc.subject.other | Bacteriology | |
dc.subject.other | Mycobacterium tuberculosis | |
dc.subject.other | Antigen | |
dc.subject.other | Glycoprotein | |
dc.subject.other | Pichia pastoris | |
dc.subject.other | PstS-1 | |
dc.subject.other | Komagataella phaffii | |
dc.title | Recombinant O-mannosylated protein production (PstS-1) from Mycobacterium tuberculosis in Pichia pastoris (Komagataella phaffii) as a tool to study tuberculosis infection. | |
dc.type | Artículo | |
dc.type | publishedVersion | |
dcterms.bibliographicCitation | Microbial Cell Factories (1475-2859 ) 18(1), 1-19 (2019) | |
dcterms.creator | Bando Campos, Carlos Giroshi::cvu::366135 | |
dcterms.creator | Juarez Lopez, Daniel::cvu::388590 | |
dcterms.creator | Roman Gonzalez, Sergio Agustin::cvu::165425 | |
dcterms.creator | Castillo Rodal, Antonia Isabel::cvu::36850 | |
dcterms.creator | Olvera Carranza, Clarita::cvu::21261 | |
dcterms.creator | Lopez Vidal, Yolanda::cvu::5656 | |
dcterms.creator | Arreguin Espinosa de los Monteros, Roberto::cvu::7506 | |
dcterms.creator | Espitia Pinzon, Clara Ines::cvu::11232 | |
dcterms.creator | Trujillo Roldan, Mauricio Alberto::cvu::201926 | |
dcterms.creator | Valdez Cruz, Norma Adriana::cvu::90992 | |
dc.identifier.doi | 10.1186/s12934-019-1059-3 | |
dc.relation.ispartofjournal | https://microbialcellfactories.biomedcentral.com/articles?query=&volume=18&searchType=&tab=keyword | |
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