Fate of the UPR marker protein Kar2/Bip and autophagic processes in fed-batch cultures of secretory insulin precursor producing Pichia pastoris View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2018-12

AUTHORS

Gustavo Roth, Ana Letícia Vanz, Heinrich Lünsdorf, Manfred Nimtz, Ursula Rinas

ABSTRACT

BACKGROUND: Secretory recombinant protein production with Pichia (syn. Komagataella) pastoris is commonly associated with the induction of an unfolded protein response (UPR) usually apparent through increased intracellular levels of endoplasmic reticulum (ER) resident chaperones such as Kar2/Bip. During methanol-induced secretory production of an insulin precursor (IP) under industrially relevant fed-batch conditions the initially high level of intracellular Kar2/Bip after batch growth on glycerol unexpectedly declined in the following methanol fed-batch phase misleadingly suggesting that IP production had a low impact on UPR activation. RESULTS: Analysis of the protein production independent level of Kar2/Bip revealed that high Kar2/Bip levels were reached in the exponential growth phase of glycerol batch cultures followed by a strong decline of Kar2/Bip during entry into stationary phase. Ultra-structural cell morphology studies revealed autophagic processes (e.g. ER phagy) at the end of the glycerol batch phase most likely responsible for the degradation of ER resident chaperones such as Kar2/Bip. The pre-induction level of Kar2/Bip did not affect the IP secretion efficiency in the subsequent methanol-induced IP production phase. During growth on methanol intracellular Kar2/Bip levels declined in IP producing and non-producing host cells. However, extracellular accumulation of Kar2/Bip was observed in IP-producing cultures but not in non-producing controls. Most importantly, the majority of the extracellular Kar2/Bip accumulated in the culture supernatant of IP producing cells as truncated protein (approx. 35 kDa). CONCLUSIONS: Rapid growth leads to higher basal levels of the major UPR marker protein Kar2/Bip independent of recombinant protein production. Entry into stationary phase or slower growth on poorer substrate, e.g. methanol, leads to a lower basal Kar2/Bip level. Methanol-induced secretory IP production elicits a strong UPR activation which counteracts the reduced UPR during slow growth on methanol. The major ER chaperone Kar2/Bip is found together with recombinant IP in the culture medium where full-length Kar2/Bip accumulates in addition to large amounts of truncated Kar2/Bip. Thus, for judging UPR activating properties of the produced protein it is important to additionally analyze the medium not only for intact Kar2/Bip but also for truncated versions of this UPR reporter protein. More... »

PAGES

123

References to SciGraph publications

  • 2013-11. Investigating the physiological response of Pichia (Komagataella) pastoris GS115 to the heterologous expression of misfolded proteins using chemostat cultures in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2010-12. The response to unfolded protein is involved in osmotolerance of Pichia pastoris in BMC GENOMICS
  • 2015-04. Quo vadis? The challenges of recombinant protein folding and secretion in Pichia pastoris in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2007-02. Cooverexpression of chaperones for enhanced secretion of a single-chain antibody fragment in Pichia pastoris in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2014-12. High-temperature cultivation of recombinant Pichia pastorisincreases endoplasmic reticulum stress and decreases production of human interleukin-10 in MICROBIAL CELL FACTORIES
  • 2002-06. The unfolded protein response in nutrient sensing and differentiation in NATURE REVIEWS MOLECULAR CELL BIOLOGY
  • 2010-12. The HAC1 gene from Pichia pastoris: characterization and effect of its overexpression on the production of secreted, surface displayed and membrane proteins in MICROBIAL CELL FACTORIES
  • 2012-03. Intracellular interactome of secreted antibody Fab fragment in Pichia pastoris reveals its routes of secretion and degradation in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2009-12. Simple high-cell density fed-batch technique for high-level recombinant protein production with Pichia pastoris: Application to intracellular production of Hepatitis B surface antigen in MICROBIAL CELL FACTORIES
  • 2014-12. Decrease of UPR- and ERAD-related proteins in Pichia pastoris during methanol-induced secretory insulin precursor production in controlled fed-batch cultures in MICROBIAL CELL FACTORIES
  • 2009-05. Functions and pathologies of BiP and its interaction partners in CELLULAR AND MOLECULAR LIFE SCIENCES
  • 2016-06. Effect of cooperation of chaperones and gene dosage on the expression of porcine PGLYRP-1 in Pichia pastoris in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2012-12. Physiological response of Pichia pastoris GS115 to methanol-induced high level production of the Hepatitis B surface antigen: catabolic adaptation, stress responses, and autophagic processes in MICROBIAL CELL FACTORIES
  • 2011-12. Virus-like particle production with yeast: ultrastructural and immunocytochemical insights into Pichia pastoris producing high levels of the Hepatitis B surface antigen in MICROBIAL CELL FACTORIES
  • 2010-12. Application of simple fed-batch technique to high-level secretory production of insulin precursor using Pichia pastoris with subsequent purification and conversion to human insulin in MICROBIAL CELL FACTORIES
  • 2007-12. Monitoring of transcriptional regulation in Pichia pastoris under protein production conditions in BMC GENOMICS
  • 2011-02. Understanding the effect of foreign gene dosage on the physiology of Pichia pastoris by transcriptional analysis of key genes in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2007-10. Transcriptional response of P. pastoris in fed-batch cultivations to Rhizopus oryzae lipase production reveals UPR induction in MICROBIAL CELL FACTORIES
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1186/s12934-018-0970-3

    DOI

    http://dx.doi.org/10.1186/s12934-018-0970-3

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1106081318

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/30092809


    Indexing Status Check whether this publication has been indexed by Scopus and Web Of Science using the SN Indexing Status Tool
    Incoming Citations Browse incoming citations for this publication using opencitations.net

    JSON-LD is the canonical representation for SciGraph data.

    TIP: You can open this SciGraph record using an external JSON-LD service: JSON-LD Playground Google SDTT

    [
      {
        "@context": "https://springernature.github.io/scigraph/jsonld/sgcontext.json", 
        "about": [
          {
            "id": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/0601", 
            "inDefinedTermSet": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/", 
            "name": "Biochemistry and Cell Biology", 
            "type": "DefinedTerm"
          }, 
          {
            "id": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/06", 
            "inDefinedTermSet": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/", 
            "name": "Biological Sciences", 
            "type": "DefinedTerm"
          }
        ], 
        "author": [
          {
            "affiliation": {
              "alternateName": "University of Hannover", 
              "id": "https://www.grid.ac/institutes/grid.9122.8", 
              "name": [
                "Technical Chemistry\u2013Life Science, Leibniz University of Hannover, Hannover, Germany"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Roth", 
            "givenName": "Gustavo", 
            "id": "sg:person.07565776255.02", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.07565776255.02"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "University of Hannover", 
              "id": "https://www.grid.ac/institutes/grid.9122.8", 
              "name": [
                "Technical Chemistry\u2013Life Science, Leibniz University of Hannover, Hannover, Germany"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Vanz", 
            "givenName": "Ana Let\u00edcia", 
            "id": "sg:person.01110235364.69", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01110235364.69"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "name": [
                "Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124, Brunswick, Germany"
              ], 
              "type": "Organization"
            }, 
            "familyName": "L\u00fcnsdorf", 
            "givenName": "Heinrich", 
            "id": "sg:person.01235676637.55", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01235676637.55"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "name": [
                "Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124, Brunswick, Germany"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Nimtz", 
            "givenName": "Manfred", 
            "id": "sg:person.01310136664.45", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01310136664.45"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "University of Hannover", 
              "id": "https://www.grid.ac/institutes/grid.9122.8", 
              "name": [
                "Technical Chemistry\u2013Life Science, Leibniz University of Hannover, Hannover, Germany", 
                "Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124, Brunswick, Germany"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Rinas", 
            "givenName": "Ursula", 
            "id": "sg:person.01340112632.46", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01340112632.46"
            ], 
            "type": "Person"
          }
        ], 
        "citation": [
          {
            "id": "sg:pub.10.1186/1475-2859-6-21", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1006478206", 
              "https://doi.org/10.1186/1475-2859-6-21"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s00018-009-8745-y", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1007472723", 
              "https://doi.org/10.1007/s00018-009-8745-y"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "https://doi.org/10.1111/j.1574-6976.2000.tb00532.x", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1009711357"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1186/1475-2859-8-13", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1009825112", 
              "https://doi.org/10.1186/1475-2859-8-13"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "https://doi.org/10.1128/aem.71.9.5332-5340.2005", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1012961576"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s00253-013-5186-1", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1013169343", 
              "https://doi.org/10.1007/s00253-013-5186-1"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s00253-013-5186-1", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1013169343", 
              "https://doi.org/10.1007/s00253-013-5186-1"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "https://doi.org/10.1016/s0958-1669(02)00330-0", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1014666008"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s00253-006-0652-7", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1020394487", 
              "https://doi.org/10.1007/s00253-006-0652-7"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s00253-006-0652-7", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1020394487", 
              "https://doi.org/10.1007/s00253-006-0652-7"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1186/1475-2859-9-49", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1020634794", 
              "https://doi.org/10.1186/1475-2859-9-49"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "https://doi.org/10.1091/mbc.12.4.957", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1020907079"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s00253-010-2944-1", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1026437303", 
              "https://doi.org/10.1007/s00253-010-2944-1"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s00253-015-6470-z", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1026637493", 
              "https://doi.org/10.1007/s00253-015-6470-z"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s00253-012-3933-3", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1026719888", 
              "https://doi.org/10.1007/s00253-012-3933-3"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "https://doi.org/10.1101/gad.852300", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1032743486"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "https://doi.org/10.1016/j.mrfmmm.2004.06.056", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1034135579"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1186/s12934-014-0163-7", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1034173375", 
              "https://doi.org/10.1186/s12934-014-0163-7"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1186/s12934-014-0163-7", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1034173375", 
              "https://doi.org/10.1186/s12934-014-0163-7"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1186/1471-2164-8-179", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1036293890", 
              "https://doi.org/10.1186/1471-2164-8-179"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1186/1475-2859-10-48", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1037100700", 
              "https://doi.org/10.1186/1475-2859-10-48"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1038/nrm829", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1038345451", 
              "https://doi.org/10.1038/nrm829"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1038/nrm829", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1038345451", 
              "https://doi.org/10.1038/nrm829"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "https://doi.org/10.1002/biot.201300334", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1039245164"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1186/1475-2859-9-31", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1039821574", 
              "https://doi.org/10.1186/1475-2859-9-31"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "https://doi.org/10.1371/journal.pbio.0040423", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1040105961"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "https://doi.org/10.1371/journal.pone.0022692", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1040393888"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "https://doi.org/10.1002/bit.10904", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1044169922"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "https://doi.org/10.1242/jcs.154716", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1047447997"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1186/1471-2164-11-207", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1049033732", 
              "https://doi.org/10.1186/1471-2164-11-207"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "https://doi.org/10.1016/j.micres.2013.06.007", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1050957365"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "https://doi.org/10.1016/j.nbt.2014.02.009", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1051175977"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1186/1475-2859-13-23", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1051324720", 
              "https://doi.org/10.1186/1475-2859-13-23"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1186/1475-2859-11-103", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1051848306", 
              "https://doi.org/10.1186/1475-2859-11-103"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "https://doi.org/10.1016/j.jprot.2013.06.031", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1052387271"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s00253-016-7372-4", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1053003643", 
              "https://doi.org/10.1007/s00253-016-7372-4"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "https://doi.org/10.1021/pr8007623", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1056294620"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "https://doi.org/10.4161/auto.3930", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1072290190"
            ], 
            "type": "CreativeWork"
          }
        ], 
        "datePublished": "2018-12", 
        "datePublishedReg": "2018-12-01", 
        "description": "BACKGROUND: Secretory recombinant protein production with Pichia (syn. Komagataella) pastoris is commonly associated with the induction of an unfolded protein response (UPR) usually apparent through increased intracellular levels of endoplasmic reticulum (ER) resident chaperones such as Kar2/Bip. During methanol-induced secretory production of an insulin precursor (IP) under industrially relevant fed-batch conditions the initially high level of intracellular Kar2/Bip after batch growth on glycerol unexpectedly declined in the following methanol fed-batch phase misleadingly suggesting that IP production had a low impact on UPR activation.\nRESULTS: Analysis of the protein production independent level of Kar2/Bip revealed that high Kar2/Bip levels were reached in the exponential growth phase of glycerol batch cultures followed by a strong decline of Kar2/Bip during entry into stationary phase. Ultra-structural cell morphology studies revealed autophagic processes (e.g. ER phagy) at the end of the glycerol batch phase most likely responsible for the degradation of ER resident chaperones such as Kar2/Bip. The pre-induction level of Kar2/Bip did not affect the IP secretion efficiency in the subsequent methanol-induced IP production phase. During growth on methanol intracellular Kar2/Bip levels declined in IP producing and non-producing host cells. However, extracellular accumulation of Kar2/Bip was observed in IP-producing cultures but not in non-producing controls. Most importantly, the majority of the extracellular Kar2/Bip accumulated in the culture supernatant of IP producing cells as truncated protein (approx. 35\u00a0kDa).\nCONCLUSIONS: Rapid growth leads to higher basal levels of the major UPR marker protein Kar2/Bip independent of recombinant protein production. Entry into stationary phase or slower growth on poorer substrate, e.g. methanol, leads to a lower basal Kar2/Bip level. Methanol-induced secretory IP production elicits a strong UPR activation which counteracts the reduced UPR during slow growth on methanol. The major ER chaperone Kar2/Bip is found together with recombinant IP in the culture medium where full-length Kar2/Bip accumulates in addition to large amounts of truncated Kar2/Bip. Thus, for judging UPR activating properties of the produced protein it is important to additionally analyze the medium not only for intact Kar2/Bip but also for truncated versions of this UPR reporter protein.", 
        "genre": "research_article", 
        "id": "sg:pub.10.1186/s12934-018-0970-3", 
        "inLanguage": [
          "en"
        ], 
        "isAccessibleForFree": true, 
        "isPartOf": [
          {
            "id": "sg:journal.1030599", 
            "issn": [
              "1475-2859"
            ], 
            "name": "Microbial Cell Factories", 
            "type": "Periodical"
          }, 
          {
            "issueNumber": "1", 
            "type": "PublicationIssue"
          }, 
          {
            "type": "PublicationVolume", 
            "volumeNumber": "17"
          }
        ], 
        "name": "Fate of the UPR marker protein Kar2/Bip and autophagic processes in fed-batch cultures of secretory insulin precursor producing Pichia pastoris", 
        "pagination": "123", 
        "productId": [
          {
            "name": "readcube_id", 
            "type": "PropertyValue", 
            "value": [
              "47a4109fdbd1bfcdc08cc1c5fd265d604ed8e3459d3506f7569153e3d8075df3"
            ]
          }, 
          {
            "name": "pubmed_id", 
            "type": "PropertyValue", 
            "value": [
              "30092809"
            ]
          }, 
          {
            "name": "nlm_unique_id", 
            "type": "PropertyValue", 
            "value": [
              "101139812"
            ]
          }, 
          {
            "name": "doi", 
            "type": "PropertyValue", 
            "value": [
              "10.1186/s12934-018-0970-3"
            ]
          }, 
          {
            "name": "dimensions_id", 
            "type": "PropertyValue", 
            "value": [
              "pub.1106081318"
            ]
          }
        ], 
        "sameAs": [
          "https://doi.org/10.1186/s12934-018-0970-3", 
          "https://app.dimensions.ai/details/publication/pub.1106081318"
        ], 
        "sdDataset": "articles", 
        "sdDatePublished": "2019-04-10T20:55", 
        "sdLicense": "https://scigraph.springernature.com/explorer/license/", 
        "sdPublisher": {
          "name": "Springer Nature - SN SciGraph project", 
          "type": "Organization"
        }, 
        "sdSource": "s3://com-uberresearch-data-dimensions-target-20181106-alternative/cleanup/v134/2549eaecd7973599484d7c17b260dba0a4ecb94b/merge/v9/a6c9fde33151104705d4d7ff012ea9563521a3ce/jats-lookup/v90/0000000001_0000000264/records_8684_00000571.jsonl", 
        "type": "ScholarlyArticle", 
        "url": "https://link.springer.com/10.1186%2Fs12934-018-0970-3"
      }
    ]
     

    Download the RDF metadata as:  json-ld nt turtle xml License info

    HOW TO GET THIS DATA PROGRAMMATICALLY:

    JSON-LD is a popular format for linked data which is fully compatible with JSON.

    curl -H 'Accept: application/ld+json' 'https://scigraph.springernature.com/pub.10.1186/s12934-018-0970-3'

    N-Triples is a line-based linked data format ideal for batch operations.

    curl -H 'Accept: application/n-triples' 'https://scigraph.springernature.com/pub.10.1186/s12934-018-0970-3'

    Turtle is a human-readable linked data format.

    curl -H 'Accept: text/turtle' 'https://scigraph.springernature.com/pub.10.1186/s12934-018-0970-3'

    RDF/XML is a standard XML format for linked data.

    curl -H 'Accept: application/rdf+xml' 'https://scigraph.springernature.com/pub.10.1186/s12934-018-0970-3'


     

    This table displays all metadata directly associated to this object as RDF triples.

    221 TRIPLES      21 PREDICATES      63 URIs      21 LITERALS      9 BLANK NODES

    Subject Predicate Object
    1 sg:pub.10.1186/s12934-018-0970-3 schema:about anzsrc-for:06
    2 anzsrc-for:0601
    3 schema:author Nb3c34b57fdd543579f075195e4657598
    4 schema:citation sg:pub.10.1007/s00018-009-8745-y
    5 sg:pub.10.1007/s00253-006-0652-7
    6 sg:pub.10.1007/s00253-010-2944-1
    7 sg:pub.10.1007/s00253-012-3933-3
    8 sg:pub.10.1007/s00253-013-5186-1
    9 sg:pub.10.1007/s00253-015-6470-z
    10 sg:pub.10.1007/s00253-016-7372-4
    11 sg:pub.10.1038/nrm829
    12 sg:pub.10.1186/1471-2164-11-207
    13 sg:pub.10.1186/1471-2164-8-179
    14 sg:pub.10.1186/1475-2859-10-48
    15 sg:pub.10.1186/1475-2859-11-103
    16 sg:pub.10.1186/1475-2859-13-23
    17 sg:pub.10.1186/1475-2859-6-21
    18 sg:pub.10.1186/1475-2859-8-13
    19 sg:pub.10.1186/1475-2859-9-31
    20 sg:pub.10.1186/1475-2859-9-49
    21 sg:pub.10.1186/s12934-014-0163-7
    22 https://doi.org/10.1002/biot.201300334
    23 https://doi.org/10.1002/bit.10904
    24 https://doi.org/10.1016/j.jprot.2013.06.031
    25 https://doi.org/10.1016/j.micres.2013.06.007
    26 https://doi.org/10.1016/j.mrfmmm.2004.06.056
    27 https://doi.org/10.1016/j.nbt.2014.02.009
    28 https://doi.org/10.1016/s0958-1669(02)00330-0
    29 https://doi.org/10.1021/pr8007623
    30 https://doi.org/10.1091/mbc.12.4.957
    31 https://doi.org/10.1101/gad.852300
    32 https://doi.org/10.1111/j.1574-6976.2000.tb00532.x
    33 https://doi.org/10.1128/aem.71.9.5332-5340.2005
    34 https://doi.org/10.1242/jcs.154716
    35 https://doi.org/10.1371/journal.pbio.0040423
    36 https://doi.org/10.1371/journal.pone.0022692
    37 https://doi.org/10.4161/auto.3930
    38 schema:datePublished 2018-12
    39 schema:datePublishedReg 2018-12-01
    40 schema:description BACKGROUND: Secretory recombinant protein production with Pichia (syn. Komagataella) pastoris is commonly associated with the induction of an unfolded protein response (UPR) usually apparent through increased intracellular levels of endoplasmic reticulum (ER) resident chaperones such as Kar2/Bip. During methanol-induced secretory production of an insulin precursor (IP) under industrially relevant fed-batch conditions the initially high level of intracellular Kar2/Bip after batch growth on glycerol unexpectedly declined in the following methanol fed-batch phase misleadingly suggesting that IP production had a low impact on UPR activation. RESULTS: Analysis of the protein production independent level of Kar2/Bip revealed that high Kar2/Bip levels were reached in the exponential growth phase of glycerol batch cultures followed by a strong decline of Kar2/Bip during entry into stationary phase. Ultra-structural cell morphology studies revealed autophagic processes (e.g. ER phagy) at the end of the glycerol batch phase most likely responsible for the degradation of ER resident chaperones such as Kar2/Bip. The pre-induction level of Kar2/Bip did not affect the IP secretion efficiency in the subsequent methanol-induced IP production phase. During growth on methanol intracellular Kar2/Bip levels declined in IP producing and non-producing host cells. However, extracellular accumulation of Kar2/Bip was observed in IP-producing cultures but not in non-producing controls. Most importantly, the majority of the extracellular Kar2/Bip accumulated in the culture supernatant of IP producing cells as truncated protein (approx. 35 kDa). CONCLUSIONS: Rapid growth leads to higher basal levels of the major UPR marker protein Kar2/Bip independent of recombinant protein production. Entry into stationary phase or slower growth on poorer substrate, e.g. methanol, leads to a lower basal Kar2/Bip level. Methanol-induced secretory IP production elicits a strong UPR activation which counteracts the reduced UPR during slow growth on methanol. The major ER chaperone Kar2/Bip is found together with recombinant IP in the culture medium where full-length Kar2/Bip accumulates in addition to large amounts of truncated Kar2/Bip. Thus, for judging UPR activating properties of the produced protein it is important to additionally analyze the medium not only for intact Kar2/Bip but also for truncated versions of this UPR reporter protein.
    41 schema:genre research_article
    42 schema:inLanguage en
    43 schema:isAccessibleForFree true
    44 schema:isPartOf N5cb158276b1f43a3b730dffffd223c89
    45 N8c810e0ddeab4a4486c282276f501a48
    46 sg:journal.1030599
    47 schema:name Fate of the UPR marker protein Kar2/Bip and autophagic processes in fed-batch cultures of secretory insulin precursor producing Pichia pastoris
    48 schema:pagination 123
    49 schema:productId N1146fc02a6d54461b290c15bfa73edc1
    50 N2ee4f60ba323453584b5fcc9a05b96fe
    51 N826afd4f63074ef099726d8519a00d7a
    52 N89ceace8f2884474ab36f03b683d2a6a
    53 Nd8c443e8991745e8a191af697dd182ee
    54 schema:sameAs https://app.dimensions.ai/details/publication/pub.1106081318
    55 https://doi.org/10.1186/s12934-018-0970-3
    56 schema:sdDatePublished 2019-04-10T20:55
    57 schema:sdLicense https://scigraph.springernature.com/explorer/license/
    58 schema:sdPublisher N156d8d4bbef34df7b35b6ae89868285a
    59 schema:url https://link.springer.com/10.1186%2Fs12934-018-0970-3
    60 sgo:license sg:explorer/license/
    61 sgo:sdDataset articles
    62 rdf:type schema:ScholarlyArticle
    63 N1146fc02a6d54461b290c15bfa73edc1 schema:name readcube_id
    64 schema:value 47a4109fdbd1bfcdc08cc1c5fd265d604ed8e3459d3506f7569153e3d8075df3
    65 rdf:type schema:PropertyValue
    66 N156d8d4bbef34df7b35b6ae89868285a schema:name Springer Nature - SN SciGraph project
    67 rdf:type schema:Organization
    68 N2ee4f60ba323453584b5fcc9a05b96fe schema:name doi
    69 schema:value 10.1186/s12934-018-0970-3
    70 rdf:type schema:PropertyValue
    71 N58186fdc2456416197cf5250d2446df8 schema:name Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124, Brunswick, Germany
    72 rdf:type schema:Organization
    73 N5cb158276b1f43a3b730dffffd223c89 schema:volumeNumber 17
    74 rdf:type schema:PublicationVolume
    75 N826afd4f63074ef099726d8519a00d7a schema:name dimensions_id
    76 schema:value pub.1106081318
    77 rdf:type schema:PropertyValue
    78 N89ceace8f2884474ab36f03b683d2a6a schema:name nlm_unique_id
    79 schema:value 101139812
    80 rdf:type schema:PropertyValue
    81 N8c810e0ddeab4a4486c282276f501a48 schema:issueNumber 1
    82 rdf:type schema:PublicationIssue
    83 N9a110f7f5c304160abb6b9badfe2dbdf rdf:first sg:person.01310136664.45
    84 rdf:rest Na298d0b85fc34d0d8c9d5b62dedc7d64
    85 Na298d0b85fc34d0d8c9d5b62dedc7d64 rdf:first sg:person.01340112632.46
    86 rdf:rest rdf:nil
    87 Nb3c34b57fdd543579f075195e4657598 rdf:first sg:person.07565776255.02
    88 rdf:rest Ndbd07f448f3a4d07add6a68befe08df2
    89 Nd11074d72a7542cf832dcf8896f063df rdf:first sg:person.01235676637.55
    90 rdf:rest N9a110f7f5c304160abb6b9badfe2dbdf
    91 Nd5312da51bbc434d855cf09e68b5c6d6 schema:name Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124, Brunswick, Germany
    92 rdf:type schema:Organization
    93 Nd8c443e8991745e8a191af697dd182ee schema:name pubmed_id
    94 schema:value 30092809
    95 rdf:type schema:PropertyValue
    96 Ndbd07f448f3a4d07add6a68befe08df2 rdf:first sg:person.01110235364.69
    97 rdf:rest Nd11074d72a7542cf832dcf8896f063df
    98 anzsrc-for:06 schema:inDefinedTermSet anzsrc-for:
    99 schema:name Biological Sciences
    100 rdf:type schema:DefinedTerm
    101 anzsrc-for:0601 schema:inDefinedTermSet anzsrc-for:
    102 schema:name Biochemistry and Cell Biology
    103 rdf:type schema:DefinedTerm
    104 sg:journal.1030599 schema:issn 1475-2859
    105 schema:name Microbial Cell Factories
    106 rdf:type schema:Periodical
    107 sg:person.01110235364.69 schema:affiliation https://www.grid.ac/institutes/grid.9122.8
    108 schema:familyName Vanz
    109 schema:givenName Ana Letícia
    110 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01110235364.69
    111 rdf:type schema:Person
    112 sg:person.01235676637.55 schema:affiliation N58186fdc2456416197cf5250d2446df8
    113 schema:familyName Lünsdorf
    114 schema:givenName Heinrich
    115 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01235676637.55
    116 rdf:type schema:Person
    117 sg:person.01310136664.45 schema:affiliation Nd5312da51bbc434d855cf09e68b5c6d6
    118 schema:familyName Nimtz
    119 schema:givenName Manfred
    120 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01310136664.45
    121 rdf:type schema:Person
    122 sg:person.01340112632.46 schema:affiliation https://www.grid.ac/institutes/grid.9122.8
    123 schema:familyName Rinas
    124 schema:givenName Ursula
    125 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01340112632.46
    126 rdf:type schema:Person
    127 sg:person.07565776255.02 schema:affiliation https://www.grid.ac/institutes/grid.9122.8
    128 schema:familyName Roth
    129 schema:givenName Gustavo
    130 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.07565776255.02
    131 rdf:type schema:Person
    132 sg:pub.10.1007/s00018-009-8745-y schema:sameAs https://app.dimensions.ai/details/publication/pub.1007472723
    133 https://doi.org/10.1007/s00018-009-8745-y
    134 rdf:type schema:CreativeWork
    135 sg:pub.10.1007/s00253-006-0652-7 schema:sameAs https://app.dimensions.ai/details/publication/pub.1020394487
    136 https://doi.org/10.1007/s00253-006-0652-7
    137 rdf:type schema:CreativeWork
    138 sg:pub.10.1007/s00253-010-2944-1 schema:sameAs https://app.dimensions.ai/details/publication/pub.1026437303
    139 https://doi.org/10.1007/s00253-010-2944-1
    140 rdf:type schema:CreativeWork
    141 sg:pub.10.1007/s00253-012-3933-3 schema:sameAs https://app.dimensions.ai/details/publication/pub.1026719888
    142 https://doi.org/10.1007/s00253-012-3933-3
    143 rdf:type schema:CreativeWork
    144 sg:pub.10.1007/s00253-013-5186-1 schema:sameAs https://app.dimensions.ai/details/publication/pub.1013169343
    145 https://doi.org/10.1007/s00253-013-5186-1
    146 rdf:type schema:CreativeWork
    147 sg:pub.10.1007/s00253-015-6470-z schema:sameAs https://app.dimensions.ai/details/publication/pub.1026637493
    148 https://doi.org/10.1007/s00253-015-6470-z
    149 rdf:type schema:CreativeWork
    150 sg:pub.10.1007/s00253-016-7372-4 schema:sameAs https://app.dimensions.ai/details/publication/pub.1053003643
    151 https://doi.org/10.1007/s00253-016-7372-4
    152 rdf:type schema:CreativeWork
    153 sg:pub.10.1038/nrm829 schema:sameAs https://app.dimensions.ai/details/publication/pub.1038345451
    154 https://doi.org/10.1038/nrm829
    155 rdf:type schema:CreativeWork
    156 sg:pub.10.1186/1471-2164-11-207 schema:sameAs https://app.dimensions.ai/details/publication/pub.1049033732
    157 https://doi.org/10.1186/1471-2164-11-207
    158 rdf:type schema:CreativeWork
    159 sg:pub.10.1186/1471-2164-8-179 schema:sameAs https://app.dimensions.ai/details/publication/pub.1036293890
    160 https://doi.org/10.1186/1471-2164-8-179
    161 rdf:type schema:CreativeWork
    162 sg:pub.10.1186/1475-2859-10-48 schema:sameAs https://app.dimensions.ai/details/publication/pub.1037100700
    163 https://doi.org/10.1186/1475-2859-10-48
    164 rdf:type schema:CreativeWork
    165 sg:pub.10.1186/1475-2859-11-103 schema:sameAs https://app.dimensions.ai/details/publication/pub.1051848306
    166 https://doi.org/10.1186/1475-2859-11-103
    167 rdf:type schema:CreativeWork
    168 sg:pub.10.1186/1475-2859-13-23 schema:sameAs https://app.dimensions.ai/details/publication/pub.1051324720
    169 https://doi.org/10.1186/1475-2859-13-23
    170 rdf:type schema:CreativeWork
    171 sg:pub.10.1186/1475-2859-6-21 schema:sameAs https://app.dimensions.ai/details/publication/pub.1006478206
    172 https://doi.org/10.1186/1475-2859-6-21
    173 rdf:type schema:CreativeWork
    174 sg:pub.10.1186/1475-2859-8-13 schema:sameAs https://app.dimensions.ai/details/publication/pub.1009825112
    175 https://doi.org/10.1186/1475-2859-8-13
    176 rdf:type schema:CreativeWork
    177 sg:pub.10.1186/1475-2859-9-31 schema:sameAs https://app.dimensions.ai/details/publication/pub.1039821574
    178 https://doi.org/10.1186/1475-2859-9-31
    179 rdf:type schema:CreativeWork
    180 sg:pub.10.1186/1475-2859-9-49 schema:sameAs https://app.dimensions.ai/details/publication/pub.1020634794
    181 https://doi.org/10.1186/1475-2859-9-49
    182 rdf:type schema:CreativeWork
    183 sg:pub.10.1186/s12934-014-0163-7 schema:sameAs https://app.dimensions.ai/details/publication/pub.1034173375
    184 https://doi.org/10.1186/s12934-014-0163-7
    185 rdf:type schema:CreativeWork
    186 https://doi.org/10.1002/biot.201300334 schema:sameAs https://app.dimensions.ai/details/publication/pub.1039245164
    187 rdf:type schema:CreativeWork
    188 https://doi.org/10.1002/bit.10904 schema:sameAs https://app.dimensions.ai/details/publication/pub.1044169922
    189 rdf:type schema:CreativeWork
    190 https://doi.org/10.1016/j.jprot.2013.06.031 schema:sameAs https://app.dimensions.ai/details/publication/pub.1052387271
    191 rdf:type schema:CreativeWork
    192 https://doi.org/10.1016/j.micres.2013.06.007 schema:sameAs https://app.dimensions.ai/details/publication/pub.1050957365
    193 rdf:type schema:CreativeWork
    194 https://doi.org/10.1016/j.mrfmmm.2004.06.056 schema:sameAs https://app.dimensions.ai/details/publication/pub.1034135579
    195 rdf:type schema:CreativeWork
    196 https://doi.org/10.1016/j.nbt.2014.02.009 schema:sameAs https://app.dimensions.ai/details/publication/pub.1051175977
    197 rdf:type schema:CreativeWork
    198 https://doi.org/10.1016/s0958-1669(02)00330-0 schema:sameAs https://app.dimensions.ai/details/publication/pub.1014666008
    199 rdf:type schema:CreativeWork
    200 https://doi.org/10.1021/pr8007623 schema:sameAs https://app.dimensions.ai/details/publication/pub.1056294620
    201 rdf:type schema:CreativeWork
    202 https://doi.org/10.1091/mbc.12.4.957 schema:sameAs https://app.dimensions.ai/details/publication/pub.1020907079
    203 rdf:type schema:CreativeWork
    204 https://doi.org/10.1101/gad.852300 schema:sameAs https://app.dimensions.ai/details/publication/pub.1032743486
    205 rdf:type schema:CreativeWork
    206 https://doi.org/10.1111/j.1574-6976.2000.tb00532.x schema:sameAs https://app.dimensions.ai/details/publication/pub.1009711357
    207 rdf:type schema:CreativeWork
    208 https://doi.org/10.1128/aem.71.9.5332-5340.2005 schema:sameAs https://app.dimensions.ai/details/publication/pub.1012961576
    209 rdf:type schema:CreativeWork
    210 https://doi.org/10.1242/jcs.154716 schema:sameAs https://app.dimensions.ai/details/publication/pub.1047447997
    211 rdf:type schema:CreativeWork
    212 https://doi.org/10.1371/journal.pbio.0040423 schema:sameAs https://app.dimensions.ai/details/publication/pub.1040105961
    213 rdf:type schema:CreativeWork
    214 https://doi.org/10.1371/journal.pone.0022692 schema:sameAs https://app.dimensions.ai/details/publication/pub.1040393888
    215 rdf:type schema:CreativeWork
    216 https://doi.org/10.4161/auto.3930 schema:sameAs https://app.dimensions.ai/details/publication/pub.1072290190
    217 rdf:type schema:CreativeWork
    218 https://www.grid.ac/institutes/grid.9122.8 schema:alternateName University of Hannover
    219 schema:name Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124, Brunswick, Germany
    220 Technical Chemistry–Life Science, Leibniz University of Hannover, Hannover, Germany
    221 rdf:type schema:Organization
     




    Preview window. Press ESC to close (or click here)


    ...