Evidence map›Paper›PMID 30660186›Full record

ArticleMicrobial cell factories2019

Recombinant O-mannosylated protein production (PstS-1) from Mycobacterium tuberculosis in Pichia pastoris (Komagataella phaffii) as a tool to study tuberculosis infection.

Giroshi Bando-Campos, Daniel Juárez-López, Sergio A Román-González, Antonia I Castillo-Rodal, Clarita Olvera, Yolanda López-Vidal, Roberto Arreguín-Espinosa, Clara Espitia, Mauricio A Trujillo-Roldán, Norma A Valdez-Cruz

Abstract read
In one paragraph

Article in Microbial cell factories, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.

0numbers the graph read from it
0cells of the map it votes in
5citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

5 citing papers in PubMed.

  1. Article
  2. mtTB: A Web-Based R/Shiny App for Pulmonary Tuberculosis Screening.Frontiers in cellular and infection microbiology · 2022
    Article
  3. Review
  4. Article
  5. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

10 authors.

Giroshi Bando-CamposPrograma de Investigación de Producción de Biomoléculas, Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, AP. 70228, CP. 04510, Ciudad de México, Mexico.
Daniel Juárez-LópezPrograma de Investigación de Producción de Biomoléculas, Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, AP. 70228, CP. 04510, Ciudad de México, Mexico.
Sergio A Román-GonzálezUnidad de Proteómica, Instituto Nacional de Medicina Genómica (INMEGEN), Periférico Sur 4809, Col. Arenal Tepepan, Tlalpan, C.P. 14610, Ciudad de México, Mexico.
Antonia I Castillo-RodalPrograma de Inmunología Molecular Microbiana, Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), 04510, Ciudad de México, Mexico.
Clarita OlveraDepartamento de Ingeniería Celular y Biocatálisis, Instituto de Biotecnología UNAM, Av. Universidad 2001 Chamilpa, Cuernavaca, Morelos, Mexico.
Yolanda López-VidalPrograma de Inmunología Molecular Microbiana, Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), 04510, Ciudad de México, Mexico.
Roberto Arreguín-EspinosaDepartamento de Química de Biomacromoléculas, Instituto de Química, Universidad Nacional Autónoma de México, Av. Universidad 3000, Ciudad Universitaria, Apdo, Postal 70250, C.P. 04510, México City, Mexico.
Clara EspitiaDepartamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Ciudad de México, Mexico.
Mauricio A Trujillo-RoldánPrograma de Investigación de Producción de Biomoléculas, Unidad de Bioprocesos, Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, AP. 70228, CP. 04510, Ciudad de México, Mexico.
Norma A Valdez-CruzPrograma de Investigación de Producción de Biomoléculas, Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, AP. 70228, CP. 04510, Ciudad de México, Mexico. adri@biomedicas.unam.mx.ORCID http://orcid.org/0000-0002-8581-1173

Funding

Consejo Nacional de Ciencia y Tecnología 220795Consejo Nacional de Ciencia y Tecnología 247473Consejo Nacional de Ciencia y Tecnología 288329Consejo Nacional de Ciencia y Tecnología 366135Consejo Nacional de Ciencia y Tecnología 388590Dirección General de Asuntos del Personal Académico, Universidad Nacional Autónoma de México IN-208415Dirección General de Asuntos del Personal Académico, Universidad Nacional Autónoma de México IN-209113
6 · The paper itself

Abstract

backgroundPichia 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.

resultsThe 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 α-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.

conclusionsrPstS-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.

Indexed as

Aldehyde OxidaseAntibodies, BacterialATP-Binding Cassette TransportersBacterial ProteinsBioreactorsCircular DichroismElectrophoresis, Polyacrylamide GelGlycosylationHumansMycobacterium tuberculosisPichiaPlasmidsPromoter Regions, GeneticProtein Structure, SecondaryRecombinant ProteinsSpectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationAldehyde OxidaseAntibodies, BacterialATP-Binding Cassette TransportersBacterial ProteinsPstS-1 protein, Mycobacterium tuberculosisRecombinant ProteinsAntigenGlycoproteinKomagataella phaffiiMycobacterium tuberculosisO-mannosylationPichia pastorisPstS-1

Identifiers

PMID30660186
PMCPMC6339365

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.