Prokaryotic soluble expression and purification of bioactive human fibroblast growth factor 21 using maltose-binding protein View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2017-11-23

AUTHORS

Anh Ngoc Nguyen, Jung-A Song, Minh Tan Nguyen, Bich Hang, Grace G. Kwon, Sang Su Park, Jiwon Yoo, Jaepyeong Jang, Jonghwa Jin, Mark J. Osborn, Yeon Jin Jang, Thu Trang Thi Vu, Heung-Bum Oh, Han Choe

ABSTRACT

Human fibroblast growth factor 21 (hFGF21) has been characterized as an important regulator of glucose and lipid metabolism homeostasis. Here, to produce hFGF21 efficiently in Escherichia coli, the expression and solubility of hFGF21 were tested and optimised by fusing the protein with one of eight tags: hexahistidine (His6), thioredoxin (Trx), small ubiquitin-related modifier (Sumo), glutathione S-transferase (GST), maltose-binding protein (MBP), N-utilisation substance protein A (NusA), human protein disulphide isomerase (PDI), and the b'a' domain of PDI (PDIb'a'). Each tag increased solubility of the protein when the expression temperature was 18°C. Unlike many other tags that were tested, MBP significantly enhanced the solubility of the protein also in the culture condition at 37°C. Thus, the MBP-hFGF21 construct was further pursued for optimisation of affinity chromatography purification. After tag removal, 8.1 mg of pure hFGF21 was obtained as a final product from 500 mL of starting culture. The protein was then characterised by mass spectroscopy and an in vitro functional assay using NIH-3T3 cells transfected with a β-klotho reporter gene. These characteristics are similar to those of commercial hFGF21. Thus, the MBP tag is useful for efficient prokaryotic production and purification of bioactive hFGF21. More... »

PAGES

16139

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/s41598-017-16167-x

DOI

http://dx.doi.org/10.1038/s41598-017-16167-x

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1092756173

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/29170489


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/2008/06", 
        "inDefinedTermSet": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/", 
        "name": "Biological Sciences", 
        "type": "DefinedTerm"
      }, 
      {
        "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"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Escherichia coli", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Fibroblast Growth Factors", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Humans", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Maltose-Binding Proteins", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Prokaryotic Cells", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Protein Disulfide-Isomerases", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Recombinant Fusion Proteins", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Thioredoxins", 
        "type": "DefinedTerm"
      }
    ], 
    "author": [
      {
        "affiliation": {
          "alternateName": "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea", 
          "id": "http://www.grid.ac/institutes/grid.413967.e", 
          "name": [
            "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Nguyen", 
        "givenName": "Anh Ngoc", 
        "id": "sg:person.01011330553.01", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01011330553.01"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea", 
          "id": "http://www.grid.ac/institutes/grid.413967.e", 
          "name": [
            "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Song", 
        "givenName": "Jung-A", 
        "id": "sg:person.01020172546.14", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01020172546.14"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea", 
          "id": "http://www.grid.ac/institutes/grid.413967.e", 
          "name": [
            "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Nguyen", 
        "givenName": "Minh Tan", 
        "id": "sg:person.01134421146.75", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01134421146.75"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea", 
          "id": "http://www.grid.ac/institutes/grid.413967.e", 
          "name": [
            "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Hang", 
        "givenName": "Bich", 
        "id": "sg:person.014147216547.18", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.014147216547.18"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea", 
          "id": "http://www.grid.ac/institutes/grid.413967.e", 
          "name": [
            "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Kwon", 
        "givenName": "Grace G.", 
        "id": "sg:person.013705732743.83", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.013705732743.83"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea", 
          "id": "http://www.grid.ac/institutes/grid.413967.e", 
          "name": [
            "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Park", 
        "givenName": "Sang Su", 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea", 
          "id": "http://www.grid.ac/institutes/grid.413967.e", 
          "name": [
            "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Yoo", 
        "givenName": "Jiwon", 
        "id": "sg:person.01270624653.49", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01270624653.49"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea", 
          "id": "http://www.grid.ac/institutes/grid.413967.e", 
          "name": [
            "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Jang", 
        "givenName": "Jaepyeong", 
        "id": "sg:person.014503313343.96", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.014503313343.96"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "New Drug Development Center, Osong Medical Innovation Foundation, 123 Osong Saengmyung-Ro, Cheongju, Chungcheongbuk-Do 28160 Korea", 
          "id": "http://www.grid.ac/institutes/grid.496741.9", 
          "name": [
            "New Drug Development Center, Osong Medical Innovation Foundation, 123 Osong Saengmyung-Ro, Cheongju, Chungcheongbuk-Do 28160 Korea"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Jin", 
        "givenName": "Jonghwa", 
        "id": "sg:person.0613333753.38", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0613333753.38"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Pediatrics, Division of Blood and Marrow Transplantation, Center for Genome Engineering, Stem Cell Institute, University of Minnesota, Minneapolis, MN 55455 USA", 
          "id": "http://www.grid.ac/institutes/grid.17635.36", 
          "name": [
            "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea", 
            "Department of Pediatrics, Division of Blood and Marrow Transplantation, Center for Genome Engineering, Stem Cell Institute, University of Minnesota, Minneapolis, MN 55455 USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Osborn", 
        "givenName": "Mark J.", 
        "id": "sg:person.010772005243.79", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.010772005243.79"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Physiology, Cell Dysfunction Research Center, University of Ulsan College of Medicine, Seoul, 05505 Korea", 
          "id": "http://www.grid.ac/institutes/grid.267370.7", 
          "name": [
            "Department of Physiology, Cell Dysfunction Research Center, University of Ulsan College of Medicine, Seoul, 05505 Korea"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Jang", 
        "givenName": "Yeon Jin", 
        "id": "sg:person.01361277155.62", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01361277155.62"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea", 
          "id": "http://www.grid.ac/institutes/grid.413967.e", 
          "name": [
            "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Vu", 
        "givenName": "Thu Trang Thi", 
        "id": "sg:person.01250647546.34", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01250647546.34"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, 05505 Korea", 
          "id": "http://www.grid.ac/institutes/grid.413967.e", 
          "name": [
            "Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, 05505 Korea"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Oh", 
        "givenName": "Heung-Bum", 
        "id": "sg:person.01032313303.24", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01032313303.24"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea", 
          "id": "http://www.grid.ac/institutes/grid.413967.e", 
          "name": [
            "Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Choe", 
        "givenName": "Han", 
        "id": "sg:person.01034610220.06", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01034610220.06"
        ], 
        "type": "Person"
      }
    ], 
    "citation": [
      {
        "id": "sg:pub.10.1186/1472-6750-10-14", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1052630425", 
          "https://doi.org/10.1186/1472-6750-10-14"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/s00253-011-3427-8", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1034388615", 
          "https://doi.org/10.1007/s00253-011-3427-8"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/978-1-59745-196-3_11", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1051756381", 
          "https://doi.org/10.1007/978-1-59745-196-3_11"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/b93995", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1009557486", 
          "https://doi.org/10.1007/b93995"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1186/1475-2859-4-1", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1048360411", 
          "https://doi.org/10.1186/1475-2859-4-1"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/s12010-013-0140-3", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1030995382", 
          "https://doi.org/10.1007/s12010-013-0140-3"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/s11033-014-3812-3", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1026183238", 
          "https://doi.org/10.1007/s11033-014-3812-3"
        ], 
        "type": "CreativeWork"
      }
    ], 
    "datePublished": "2017-11-23", 
    "datePublishedReg": "2017-11-23", 
    "description": "Human fibroblast growth factor 21 (hFGF21) has been characterized as an important regulator of glucose and lipid metabolism homeostasis. Here, to produce hFGF21 efficiently in Escherichia coli, the expression and solubility of hFGF21 were tested and optimised by fusing the protein with one of eight tags: hexahistidine (His6), thioredoxin (Trx), small ubiquitin-related modifier (Sumo), glutathione S-transferase (GST), maltose-binding protein (MBP), N-utilisation substance protein A (NusA), human protein disulphide isomerase (PDI), and the b'a' domain of PDI (PDIb'a'). Each tag increased solubility of the protein when the expression temperature was 18\u00b0C. Unlike many other tags that were tested, MBP significantly enhanced the solubility of the protein also in the culture condition at 37\u00b0C. Thus, the MBP-hFGF21 construct was further pursued for optimisation of affinity chromatography purification. After tag removal, 8.1\u2009mg of pure hFGF21 was obtained as a final product from 500\u2009mL of starting culture. The protein was then characterised by mass spectroscopy and an in vitro functional assay using NIH-3T3 cells transfected with a \u03b2-klotho reporter gene. These characteristics are similar to those of commercial hFGF21. Thus, the MBP tag is useful for efficient prokaryotic production and purification of bioactive hFGF21.", 
    "genre": "article", 
    "id": "sg:pub.10.1038/s41598-017-16167-x", 
    "inLanguage": "en", 
    "isAccessibleForFree": true, 
    "isPartOf": [
      {
        "id": "sg:journal.1045337", 
        "issn": [
          "2045-2322"
        ], 
        "name": "Scientific Reports", 
        "publisher": "Springer Nature", 
        "type": "Periodical"
      }, 
      {
        "issueNumber": "1", 
        "type": "PublicationIssue"
      }, 
      {
        "type": "PublicationVolume", 
        "volumeNumber": "7"
      }
    ], 
    "keywords": [
      "protein disulfide isomerase", 
      "maltose binding protein", 
      "human protein disulfide isomerase", 
      "small ubiquitin-related modifier", 
      "NIH 3T3 cells", 
      "human fibroblast growth factor 21", 
      "disulfide isomerase", 
      "MBP tag", 
      "reporter gene", 
      "expression temperature", 
      "lipid metabolism homeostasis", 
      "soluble expression", 
      "affinity chromatography purification", 
      "important regulator", 
      "Escherichia coli", 
      "prokaryotic production", 
      "protein", 
      "glutathione S", 
      "fibroblast growth factor 21", 
      "tag removal", 
      "protein A", 
      "growth factor 21", 
      "culture conditions", 
      "metabolism homeostasis", 
      "factor 21", 
      "tags", 
      "purification", 
      "expression", 
      "b'a' domain", 
      "chromatography purification", 
      "thioredoxin", 
      "isomerase", 
      "genes", 
      "domain", 
      "regulator", 
      "hexahistidine", 
      "homeostasis", 
      "coli", 
      "transferase", 
      "cells", 
      "final product", 
      "production", 
      "spectroscopy", 
      "constructs", 
      "modifiers", 
      "culture", 
      "solubility", 
      "mass spectroscopy", 
      "glucose", 
      "products", 
      "optimization", 
      "temperature", 
      "conditions", 
      "mL", 
      "removal", 
      "characteristics", 
      "hFGF21", 
      "solubility of hFGF21", 
      "ubiquitin-related modifier", 
      "utilisation substance protein A", 
      "substance protein A", 
      "MBP-hFGF21 construct", 
      "pure hFGF21", 
      "klotho reporter gene", 
      "commercial hFGF21", 
      "efficient prokaryotic production", 
      "bioactive hFGF21", 
      "Prokaryotic soluble expression", 
      "bioactive human fibroblast growth factor 21"
    ], 
    "name": "Prokaryotic soluble expression and purification of bioactive human fibroblast growth factor 21 using maltose-binding protein", 
    "pagination": "16139", 
    "productId": [
      {
        "name": "dimensions_id", 
        "type": "PropertyValue", 
        "value": [
          "pub.1092756173"
        ]
      }, 
      {
        "name": "doi", 
        "type": "PropertyValue", 
        "value": [
          "10.1038/s41598-017-16167-x"
        ]
      }, 
      {
        "name": "pubmed_id", 
        "type": "PropertyValue", 
        "value": [
          "29170489"
        ]
      }
    ], 
    "sameAs": [
      "https://doi.org/10.1038/s41598-017-16167-x", 
      "https://app.dimensions.ai/details/publication/pub.1092756173"
    ], 
    "sdDataset": "articles", 
    "sdDatePublished": "2021-12-01T19:40", 
    "sdLicense": "https://scigraph.springernature.com/explorer/license/", 
    "sdPublisher": {
      "name": "Springer Nature - SN SciGraph project", 
      "type": "Organization"
    }, 
    "sdSource": "s3://com-springernature-scigraph/baseset/20211201/entities/gbq_results/article/article_743.jsonl", 
    "type": "ScholarlyArticle", 
    "url": "https://doi.org/10.1038/s41598-017-16167-x"
  }
]
 

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.10.1038/s41598-017-16167-x'

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

curl -H 'Accept: application/n-triples' 'https://scigraph.springernature.com/pub.10.1038/s41598-017-16167-x'

Turtle is a human-readable linked data format.

curl -H 'Accept: text/turtle' 'https://scigraph.springernature.com/pub.10.1038/s41598-017-16167-x'

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

curl -H 'Accept: application/rdf+xml' 'https://scigraph.springernature.com/pub.10.1038/s41598-017-16167-x'


 

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

292 TRIPLES      22 PREDICATES      110 URIs      95 LITERALS      15 BLANK NODES

Subject Predicate Object
1 sg:pub.10.1038/s41598-017-16167-x schema:about N1ff87babfe0141adbe6c97fa21374f08
2 N5800834837ef484998d7b752ee30ee70
3 N7f5a1879a9bc4693bfc33fe29521137f
4 N907b416387544441a51fb7048a787b58
5 N91a43181206e4ab59aa8bcceffb6edfc
6 Na41bde0e46e5455086dd1408f8e1d03b
7 Nc6437566a6c84607a8f06325d55322e4
8 Ne8d4d50958424512b417ef1c94cae79c
9 anzsrc-for:06
10 anzsrc-for:0601
11 schema:author N383f06023f564bc7b74cb5c55657e044
12 schema:citation sg:pub.10.1007/978-1-59745-196-3_11
13 sg:pub.10.1007/b93995
14 sg:pub.10.1007/s00253-011-3427-8
15 sg:pub.10.1007/s11033-014-3812-3
16 sg:pub.10.1007/s12010-013-0140-3
17 sg:pub.10.1186/1472-6750-10-14
18 sg:pub.10.1186/1475-2859-4-1
19 schema:datePublished 2017-11-23
20 schema:datePublishedReg 2017-11-23
21 schema:description Human fibroblast growth factor 21 (hFGF21) has been characterized as an important regulator of glucose and lipid metabolism homeostasis. Here, to produce hFGF21 efficiently in Escherichia coli, the expression and solubility of hFGF21 were tested and optimised by fusing the protein with one of eight tags: hexahistidine (His6), thioredoxin (Trx), small ubiquitin-related modifier (Sumo), glutathione S-transferase (GST), maltose-binding protein (MBP), N-utilisation substance protein A (NusA), human protein disulphide isomerase (PDI), and the b'a' domain of PDI (PDIb'a'). Each tag increased solubility of the protein when the expression temperature was 18°C. Unlike many other tags that were tested, MBP significantly enhanced the solubility of the protein also in the culture condition at 37°C. Thus, the MBP-hFGF21 construct was further pursued for optimisation of affinity chromatography purification. After tag removal, 8.1 mg of pure hFGF21 was obtained as a final product from 500 mL of starting culture. The protein was then characterised by mass spectroscopy and an in vitro functional assay using NIH-3T3 cells transfected with a β-klotho reporter gene. These characteristics are similar to those of commercial hFGF21. Thus, the MBP tag is useful for efficient prokaryotic production and purification of bioactive hFGF21.
22 schema:genre article
23 schema:inLanguage en
24 schema:isAccessibleForFree true
25 schema:isPartOf N003734b84e0a44e1b06e99e139b3f253
26 N430f5ffa2b724d74b578413aeaf5525e
27 sg:journal.1045337
28 schema:keywords Escherichia coli
29 MBP tag
30 MBP-hFGF21 construct
31 NIH 3T3 cells
32 Prokaryotic soluble expression
33 affinity chromatography purification
34 b'a' domain
35 bioactive hFGF21
36 bioactive human fibroblast growth factor 21
37 cells
38 characteristics
39 chromatography purification
40 coli
41 commercial hFGF21
42 conditions
43 constructs
44 culture
45 culture conditions
46 disulfide isomerase
47 domain
48 efficient prokaryotic production
49 expression
50 expression temperature
51 factor 21
52 fibroblast growth factor 21
53 final product
54 genes
55 glucose
56 glutathione S
57 growth factor 21
58 hFGF21
59 hexahistidine
60 homeostasis
61 human fibroblast growth factor 21
62 human protein disulfide isomerase
63 important regulator
64 isomerase
65 klotho reporter gene
66 lipid metabolism homeostasis
67 mL
68 maltose binding protein
69 mass spectroscopy
70 metabolism homeostasis
71 modifiers
72 optimization
73 production
74 products
75 prokaryotic production
76 protein
77 protein A
78 protein disulfide isomerase
79 pure hFGF21
80 purification
81 regulator
82 removal
83 reporter gene
84 small ubiquitin-related modifier
85 solubility
86 solubility of hFGF21
87 soluble expression
88 spectroscopy
89 substance protein A
90 tag removal
91 tags
92 temperature
93 thioredoxin
94 transferase
95 ubiquitin-related modifier
96 utilisation substance protein A
97 schema:name Prokaryotic soluble expression and purification of bioactive human fibroblast growth factor 21 using maltose-binding protein
98 schema:pagination 16139
99 schema:productId N12dc2297af0a45f1a1c119659ecef902
100 N2b70df3a3d8f4fd5b4f243aedcda0661
101 N45d3bc88545746bdabec280059ac858e
102 schema:sameAs https://app.dimensions.ai/details/publication/pub.1092756173
103 https://doi.org/10.1038/s41598-017-16167-x
104 schema:sdDatePublished 2021-12-01T19:40
105 schema:sdLicense https://scigraph.springernature.com/explorer/license/
106 schema:sdPublisher Nf68af0fc2d79413c89b65208eeff1c2c
107 schema:url https://doi.org/10.1038/s41598-017-16167-x
108 sgo:license sg:explorer/license/
109 sgo:sdDataset articles
110 rdf:type schema:ScholarlyArticle
111 N003734b84e0a44e1b06e99e139b3f253 schema:issueNumber 1
112 rdf:type schema:PublicationIssue
113 N12dc2297af0a45f1a1c119659ecef902 schema:name doi
114 schema:value 10.1038/s41598-017-16167-x
115 rdf:type schema:PropertyValue
116 N1b418cab53ec47088c249b36a105c4e8 rdf:first sg:person.01250647546.34
117 rdf:rest N934c042b9112404caa16222489d123e2
118 N1b43560a5ac64804b730d8dfa6ea0c61 rdf:first sg:person.01270624653.49
119 rdf:rest N9101b3637c3d40c58faf0c5b45efb1ec
120 N1ff87babfe0141adbe6c97fa21374f08 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
121 schema:name Protein Disulfide-Isomerases
122 rdf:type schema:DefinedTerm
123 N2b70df3a3d8f4fd5b4f243aedcda0661 schema:name pubmed_id
124 schema:value 29170489
125 rdf:type schema:PropertyValue
126 N383f06023f564bc7b74cb5c55657e044 rdf:first sg:person.01011330553.01
127 rdf:rest N9bd33f12bfd64820908f9106cd76dce0
128 N3aa40e3e92dd48cfa4392b1cfe9e12ac rdf:first sg:person.014147216547.18
129 rdf:rest N9d1abf1040684668baabc9cf09dc2739
130 N3d74f64f8bc44582a172c89f033c5ea6 rdf:first sg:person.01034610220.06
131 rdf:rest rdf:nil
132 N430f5ffa2b724d74b578413aeaf5525e schema:volumeNumber 7
133 rdf:type schema:PublicationVolume
134 N45d3bc88545746bdabec280059ac858e schema:name dimensions_id
135 schema:value pub.1092756173
136 rdf:type schema:PropertyValue
137 N5800834837ef484998d7b752ee30ee70 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
138 schema:name Prokaryotic Cells
139 rdf:type schema:DefinedTerm
140 N64f4e1d6bbb140319fb52801a598e318 rdf:first sg:person.01134421146.75
141 rdf:rest N3aa40e3e92dd48cfa4392b1cfe9e12ac
142 N7f5a1879a9bc4693bfc33fe29521137f schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
143 schema:name Maltose-Binding Proteins
144 rdf:type schema:DefinedTerm
145 N824cb011616549f886bfb93b343824a4 rdf:first sg:person.01361277155.62
146 rdf:rest N1b418cab53ec47088c249b36a105c4e8
147 N907b416387544441a51fb7048a787b58 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
148 schema:name Thioredoxins
149 rdf:type schema:DefinedTerm
150 N9101b3637c3d40c58faf0c5b45efb1ec rdf:first sg:person.014503313343.96
151 rdf:rest N938e8a3d68ce43f4a592ae7add76a632
152 N91a43181206e4ab59aa8bcceffb6edfc schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
153 schema:name Fibroblast Growth Factors
154 rdf:type schema:DefinedTerm
155 N934c042b9112404caa16222489d123e2 rdf:first sg:person.01032313303.24
156 rdf:rest N3d74f64f8bc44582a172c89f033c5ea6
157 N938e8a3d68ce43f4a592ae7add76a632 rdf:first sg:person.0613333753.38
158 rdf:rest Nf3729a075a954ec1a5fb01fa8a48d3e3
159 N9bd33f12bfd64820908f9106cd76dce0 rdf:first sg:person.01020172546.14
160 rdf:rest N64f4e1d6bbb140319fb52801a598e318
161 N9d1abf1040684668baabc9cf09dc2739 rdf:first sg:person.013705732743.83
162 rdf:rest Nda214b7338eb4a1584a22abc0b01aa9f
163 Na3a3de5ae576438a9eab0c03a5c5ee1d schema:affiliation grid-institutes:grid.413967.e
164 schema:familyName Park
165 schema:givenName Sang Su
166 rdf:type schema:Person
167 Na41bde0e46e5455086dd1408f8e1d03b schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
168 schema:name Recombinant Fusion Proteins
169 rdf:type schema:DefinedTerm
170 Nc6437566a6c84607a8f06325d55322e4 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
171 schema:name Escherichia coli
172 rdf:type schema:DefinedTerm
173 Nda214b7338eb4a1584a22abc0b01aa9f rdf:first Na3a3de5ae576438a9eab0c03a5c5ee1d
174 rdf:rest N1b43560a5ac64804b730d8dfa6ea0c61
175 Ne8d4d50958424512b417ef1c94cae79c schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
176 schema:name Humans
177 rdf:type schema:DefinedTerm
178 Nf3729a075a954ec1a5fb01fa8a48d3e3 rdf:first sg:person.010772005243.79
179 rdf:rest N824cb011616549f886bfb93b343824a4
180 Nf68af0fc2d79413c89b65208eeff1c2c schema:name Springer Nature - SN SciGraph project
181 rdf:type schema:Organization
182 anzsrc-for:06 schema:inDefinedTermSet anzsrc-for:
183 schema:name Biological Sciences
184 rdf:type schema:DefinedTerm
185 anzsrc-for:0601 schema:inDefinedTermSet anzsrc-for:
186 schema:name Biochemistry and Cell Biology
187 rdf:type schema:DefinedTerm
188 sg:journal.1045337 schema:issn 2045-2322
189 schema:name Scientific Reports
190 schema:publisher Springer Nature
191 rdf:type schema:Periodical
192 sg:person.01011330553.01 schema:affiliation grid-institutes:grid.413967.e
193 schema:familyName Nguyen
194 schema:givenName Anh Ngoc
195 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01011330553.01
196 rdf:type schema:Person
197 sg:person.01020172546.14 schema:affiliation grid-institutes:grid.413967.e
198 schema:familyName Song
199 schema:givenName Jung-A
200 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01020172546.14
201 rdf:type schema:Person
202 sg:person.01032313303.24 schema:affiliation grid-institutes:grid.413967.e
203 schema:familyName Oh
204 schema:givenName Heung-Bum
205 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01032313303.24
206 rdf:type schema:Person
207 sg:person.01034610220.06 schema:affiliation grid-institutes:grid.413967.e
208 schema:familyName Choe
209 schema:givenName Han
210 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01034610220.06
211 rdf:type schema:Person
212 sg:person.010772005243.79 schema:affiliation grid-institutes:grid.17635.36
213 schema:familyName Osborn
214 schema:givenName Mark J.
215 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.010772005243.79
216 rdf:type schema:Person
217 sg:person.01134421146.75 schema:affiliation grid-institutes:grid.413967.e
218 schema:familyName Nguyen
219 schema:givenName Minh Tan
220 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01134421146.75
221 rdf:type schema:Person
222 sg:person.01250647546.34 schema:affiliation grid-institutes:grid.413967.e
223 schema:familyName Vu
224 schema:givenName Thu Trang Thi
225 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01250647546.34
226 rdf:type schema:Person
227 sg:person.01270624653.49 schema:affiliation grid-institutes:grid.413967.e
228 schema:familyName Yoo
229 schema:givenName Jiwon
230 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01270624653.49
231 rdf:type schema:Person
232 sg:person.01361277155.62 schema:affiliation grid-institutes:grid.267370.7
233 schema:familyName Jang
234 schema:givenName Yeon Jin
235 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01361277155.62
236 rdf:type schema:Person
237 sg:person.013705732743.83 schema:affiliation grid-institutes:grid.413967.e
238 schema:familyName Kwon
239 schema:givenName Grace G.
240 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.013705732743.83
241 rdf:type schema:Person
242 sg:person.014147216547.18 schema:affiliation grid-institutes:grid.413967.e
243 schema:familyName Hang
244 schema:givenName Bich
245 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.014147216547.18
246 rdf:type schema:Person
247 sg:person.014503313343.96 schema:affiliation grid-institutes:grid.413967.e
248 schema:familyName Jang
249 schema:givenName Jaepyeong
250 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.014503313343.96
251 rdf:type schema:Person
252 sg:person.0613333753.38 schema:affiliation grid-institutes:grid.496741.9
253 schema:familyName Jin
254 schema:givenName Jonghwa
255 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0613333753.38
256 rdf:type schema:Person
257 sg:pub.10.1007/978-1-59745-196-3_11 schema:sameAs https://app.dimensions.ai/details/publication/pub.1051756381
258 https://doi.org/10.1007/978-1-59745-196-3_11
259 rdf:type schema:CreativeWork
260 sg:pub.10.1007/b93995 schema:sameAs https://app.dimensions.ai/details/publication/pub.1009557486
261 https://doi.org/10.1007/b93995
262 rdf:type schema:CreativeWork
263 sg:pub.10.1007/s00253-011-3427-8 schema:sameAs https://app.dimensions.ai/details/publication/pub.1034388615
264 https://doi.org/10.1007/s00253-011-3427-8
265 rdf:type schema:CreativeWork
266 sg:pub.10.1007/s11033-014-3812-3 schema:sameAs https://app.dimensions.ai/details/publication/pub.1026183238
267 https://doi.org/10.1007/s11033-014-3812-3
268 rdf:type schema:CreativeWork
269 sg:pub.10.1007/s12010-013-0140-3 schema:sameAs https://app.dimensions.ai/details/publication/pub.1030995382
270 https://doi.org/10.1007/s12010-013-0140-3
271 rdf:type schema:CreativeWork
272 sg:pub.10.1186/1472-6750-10-14 schema:sameAs https://app.dimensions.ai/details/publication/pub.1052630425
273 https://doi.org/10.1186/1472-6750-10-14
274 rdf:type schema:CreativeWork
275 sg:pub.10.1186/1475-2859-4-1 schema:sameAs https://app.dimensions.ai/details/publication/pub.1048360411
276 https://doi.org/10.1186/1475-2859-4-1
277 rdf:type schema:CreativeWork
278 grid-institutes:grid.17635.36 schema:alternateName Department of Pediatrics, Division of Blood and Marrow Transplantation, Center for Genome Engineering, Stem Cell Institute, University of Minnesota, Minneapolis, MN 55455 USA
279 schema:name Department of Pediatrics, Division of Blood and Marrow Transplantation, Center for Genome Engineering, Stem Cell Institute, University of Minnesota, Minneapolis, MN 55455 USA
280 Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea
281 rdf:type schema:Organization
282 grid-institutes:grid.267370.7 schema:alternateName Department of Physiology, Cell Dysfunction Research Center, University of Ulsan College of Medicine, Seoul, 05505 Korea
283 schema:name Department of Physiology, Cell Dysfunction Research Center, University of Ulsan College of Medicine, Seoul, 05505 Korea
284 rdf:type schema:Organization
285 grid-institutes:grid.413967.e schema:alternateName Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, 05505 Korea
286 Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea
287 schema:name Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, 05505 Korea
288 Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505 Korea
289 rdf:type schema:Organization
290 grid-institutes:grid.496741.9 schema:alternateName New Drug Development Center, Osong Medical Innovation Foundation, 123 Osong Saengmyung-Ro, Cheongju, Chungcheongbuk-Do 28160 Korea
291 schema:name New Drug Development Center, Osong Medical Innovation Foundation, 123 Osong Saengmyung-Ro, Cheongju, Chungcheongbuk-Do 28160 Korea
292 rdf:type schema:Organization
 




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


...