The order of expression is a key factor in the production of active transglutaminase in Escherichia coli by co-expression with ... View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2011-12

AUTHORS

Song Liu, Dongxu Zhang, Miao Wang, Wenjing Cui, Kangkang Chen, Guocheng Du, Jian Chen, Zhemin Zhou

ABSTRACT

BACKGROUND: Streptomyces transglutaminase (TGase) is naturally synthesized as zymogen (pro-TGase), which is then processed to produce active enzyme by the removal of its N-terminal pro-peptide. This pro-peptide is found to be essential for overexpression of soluble TGase in E. coli. However, expression of pro-TGase by E. coli requires protease-mediated activation in vitro. In this study, we developed a novel co- expression method for the direct production of active TGase in E. coli. RESULTS: A TGase from S. hygroscopicus was expressed in E. coli only after fusing with the pelB signal peptide, but fusion with the signal peptide induced insoluble enzyme. Therefore, alternative protocol was designed by co-expressing the TGase and its pro-peptide as independent polypeptides under a single T7 promoter using vector pET-22b(+). Although the pro-peptide was co-expressed, the TGase fused without the signal peptide was undetectable in both soluble and insoluble fractions of the recombinant cells. Similarly, when both genes were expressed in the order of the TGase and the pro-peptide, the solubility of TGase fused with the signal peptide was not improved by the co-expression with its pro-peptide. Interestingly, active TGase was only produced by the cells in which the pro-peptide and the TGase were fused with the signal peptide and sequentially expressed. The purified recombinant and native TGase shared the similar catalytic properties. CONCLUSIONS: Our results indicated that the pro-peptide can assist correct folding of the TGase inter-molecularly in E. coli, and expression of pro-peptide prior to that of TGase was essential for the production of active TGase. The co-expression strategy based on optimizing the order of gene expression could be useful for the expression of other functional proteins that are synthesized as a precursor. More... »

PAGES

112

Identifiers

URI

http://scigraph.springernature.com/pub.10.1186/1475-2859-10-112

DOI

http://dx.doi.org/10.1186/1475-2859-10-112

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1000744820

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/22196373


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/2008/0604", 
        "inDefinedTermSet": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/", 
        "name": "Genetics", 
        "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"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Amino Acid Sequence", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Bacterial Proteins", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Enzyme Precursors", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Escherichia coli", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Molecular Sequence Data", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Protein Sorting Signals", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Protein Transport", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Recombinant Fusion Proteins", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Streptomyces", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Transglutaminases", 
        "type": "DefinedTerm"
      }
    ], 
    "author": [
      {
        "affiliation": {
          "alternateName": "Jiangnan University", 
          "id": "https://www.grid.ac/institutes/grid.258151.a", 
          "name": [
            "School of Biotechnology, Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Lihu Avenue, Wuxi, China"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Liu", 
        "givenName": "Song", 
        "id": "sg:person.0761552653.86", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0761552653.86"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Jiangnan University", 
          "id": "https://www.grid.ac/institutes/grid.258151.a", 
          "name": [
            "School of Biotechnology, Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Lihu Avenue, Wuxi, China"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Zhang", 
        "givenName": "Dongxu", 
        "id": "sg:person.01237473145.01", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01237473145.01"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Jiangnan University", 
          "id": "https://www.grid.ac/institutes/grid.258151.a", 
          "name": [
            "School of Biotechnology, Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Lihu Avenue, Wuxi, China", 
            "School of Food Science and Technology, Jiangnan University, Lihu Avenue, Wuxi, China"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Wang", 
        "givenName": "Miao", 
        "id": "sg:person.01266055146.55", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01266055146.55"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Jiangnan University", 
          "id": "https://www.grid.ac/institutes/grid.258151.a", 
          "name": [
            "School of Biotechnology, Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Lihu Avenue, Wuxi, China"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Cui", 
        "givenName": "Wenjing", 
        "id": "sg:person.01303557140.44", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01303557140.44"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Jiangnan University", 
          "id": "https://www.grid.ac/institutes/grid.258151.a", 
          "name": [
            "School of Biotechnology, Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Lihu Avenue, Wuxi, China"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Chen", 
        "givenName": "Kangkang", 
        "id": "sg:person.0775520554.94", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0775520554.94"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Jiangnan University", 
          "id": "https://www.grid.ac/institutes/grid.258151.a", 
          "name": [
            "School of Biotechnology, Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Lihu Avenue, Wuxi, China"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Du", 
        "givenName": "Guocheng", 
        "id": "sg:person.01263147440.90", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01263147440.90"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Jiangnan University", 
          "id": "https://www.grid.ac/institutes/grid.258151.a", 
          "name": [
            "School of Biotechnology, Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Lihu Avenue, Wuxi, China", 
            "State Key Laboratory of Food Science and Technology, Jiangnan University, Lihu Avenue, Wuxi, China"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Chen", 
        "givenName": "Jian", 
        "id": "sg:person.01171210206.92", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01171210206.92"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Jiangnan University", 
          "id": "https://www.grid.ac/institutes/grid.258151.a", 
          "name": [
            "School of Biotechnology, Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Lihu Avenue, Wuxi, China"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Zhou", 
        "givenName": "Zhemin", 
        "id": "sg:person.0774720425.90", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0774720425.90"
        ], 
        "type": "Person"
      }
    ], 
    "citation": [
      {
        "id": "https://doi.org/10.1271/bbb.58.88", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1005551430"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1271/bbb1961.53.2613", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1007986290"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/s00253-004-1559-9", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1009268886", 
          "https://doi.org/10.1007/s00253-004-1559-9"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1006/prep.2000.1363", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1010297488"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/s00253-008-1688-7", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1011652088", 
          "https://doi.org/10.1007/s00253-008-1688-7"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/s00253-008-1688-7", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1011652088", 
          "https://doi.org/10.1007/s00253-008-1688-7"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1128/aem.71.12.8911-8919.2005", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1017702218"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.biotechadv.2004.11.003", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1020388916"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1271/bbb.64.1263", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1021253186"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1271/bbb.68.2058", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1021692720"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.enzmictec.2008.03.003", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1022019325"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.5661/bger-26-205", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1022425337"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1046/j.1365-2958.1996.381908.x", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1023177545"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1111/j.1574-6968.2011.02387.x", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1024207652"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/s00018-010-0344-4", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1027832987", 
          "https://doi.org/10.1007/s00018-010-0344-4"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/s00018-010-0344-4", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1027832987", 
          "https://doi.org/10.1007/s00018-010-0344-4"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.febslet.2008.07.049", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1031701399"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1128/aem.69.1.358-366.2003", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1033892926"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.tibtech.2008.06.006", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1034786742"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1002/btpr.538", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1034990825"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/s00253-003-1539-5", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1040145592", 
          "https://doi.org/10.1007/s00253-003-1539-5"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.foodchem.2007.04.020", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1046226945"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1271/bbb.61.830", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1046482878"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.enzmictec.2006.10.036", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1050088038"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1111/j.1365-2958.1995.tb02423.x", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1050201312"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1021/jf703567t", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1055924478"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1021/jf703567t", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1055924478"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1021/jf8008519", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1055924931"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1021/jf8008519", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1055924931"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1101/sqb.1970.035.01.066", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1060404102"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1177/1087057109356206", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1063973629"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1177/1087057109356206", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1063973629"
        ], 
        "type": "CreativeWork"
      }
    ], 
    "datePublished": "2011-12", 
    "datePublishedReg": "2011-12-01", 
    "description": "BACKGROUND: Streptomyces transglutaminase (TGase) is naturally synthesized as zymogen (pro-TGase), which is then processed to produce active enzyme by the removal of its N-terminal pro-peptide. This pro-peptide is found to be essential for overexpression of soluble TGase in E. coli. However, expression of pro-TGase by E. coli requires protease-mediated activation in vitro. In this study, we developed a novel co- expression method for the direct production of active TGase in E. coli.\nRESULTS: A TGase from S. hygroscopicus was expressed in E. coli only after fusing with the pelB signal peptide, but fusion with the signal peptide induced insoluble enzyme. Therefore, alternative protocol was designed by co-expressing the TGase and its pro-peptide as independent polypeptides under a single T7 promoter using vector pET-22b(+). Although the pro-peptide was co-expressed, the TGase fused without the signal peptide was undetectable in both soluble and insoluble fractions of the recombinant cells. Similarly, when both genes were expressed in the order of the TGase and the pro-peptide, the solubility of TGase fused with the signal peptide was not improved by the co-expression with its pro-peptide. Interestingly, active TGase was only produced by the cells in which the pro-peptide and the TGase were fused with the signal peptide and sequentially expressed. The purified recombinant and native TGase shared the similar catalytic properties.\nCONCLUSIONS: Our results indicated that the pro-peptide can assist correct folding of the TGase inter-molecularly in E. coli, and expression of pro-peptide prior to that of TGase was essential for the production of active TGase. The co-expression strategy based on optimizing the order of gene expression could be useful for the expression of other functional proteins that are synthesized as a precursor.", 
    "genre": "research_article", 
    "id": "sg:pub.10.1186/1475-2859-10-112", 
    "inLanguage": [
      "en"
    ], 
    "isAccessibleForFree": true, 
    "isFundedItemOf": [
      {
        "id": "sg:grant.7024541", 
        "type": "MonetaryGrant"
      }, 
      {
        "id": "sg:grant.4966884", 
        "type": "MonetaryGrant"
      }, 
      {
        "id": "sg:grant.5009460", 
        "type": "MonetaryGrant"
      }, 
      {
        "id": "sg:grant.5014154", 
        "type": "MonetaryGrant"
      }
    ], 
    "isPartOf": [
      {
        "id": "sg:journal.1030599", 
        "issn": [
          "1475-2859"
        ], 
        "name": "Microbial Cell Factories", 
        "type": "Periodical"
      }, 
      {
        "issueNumber": "1", 
        "type": "PublicationIssue"
      }, 
      {
        "type": "PublicationVolume", 
        "volumeNumber": "10"
      }
    ], 
    "name": "The order of expression is a key factor in the production of active transglutaminase in Escherichia coli by co-expression with its pro-peptide", 
    "pagination": "112", 
    "productId": [
      {
        "name": "readcube_id", 
        "type": "PropertyValue", 
        "value": [
          "038c0c43ac4b42f2e7614f17771f3b70e8ac0e650e486329ec02a97384deb4ff"
        ]
      }, 
      {
        "name": "pubmed_id", 
        "type": "PropertyValue", 
        "value": [
          "22196373"
        ]
      }, 
      {
        "name": "nlm_unique_id", 
        "type": "PropertyValue", 
        "value": [
          "101139812"
        ]
      }, 
      {
        "name": "doi", 
        "type": "PropertyValue", 
        "value": [
          "10.1186/1475-2859-10-112"
        ]
      }, 
      {
        "name": "dimensions_id", 
        "type": "PropertyValue", 
        "value": [
          "pub.1000744820"
        ]
      }
    ], 
    "sameAs": [
      "https://doi.org/10.1186/1475-2859-10-112", 
      "https://app.dimensions.ai/details/publication/pub.1000744820"
    ], 
    "sdDataset": "articles", 
    "sdDatePublished": "2019-04-10T19:08", 
    "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_8678_00000509.jsonl", 
    "type": "ScholarlyArticle", 
    "url": "http://link.springer.com/10.1186%2F1475-2859-10-112"
  }
]
 

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.10.1186/1475-2859-10-112'

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

curl -H 'Accept: application/n-triples' 'https://scigraph.springernature.com/pub.10.1186/1475-2859-10-112'

Turtle is a human-readable linked data format.

curl -H 'Accept: text/turtle' 'https://scigraph.springernature.com/pub.10.1186/1475-2859-10-112'

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

curl -H 'Accept: application/rdf+xml' 'https://scigraph.springernature.com/pub.10.1186/1475-2859-10-112'


 

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

252 TRIPLES      21 PREDICATES      66 URIs      31 LITERALS      19 BLANK NODES

Subject Predicate Object
1 sg:pub.10.1186/1475-2859-10-112 schema:about N128ecf983414492daafd942f8da71df9
2 N41324025c8654ed18c5cd25e9bbc4aa0
3 N56f6e09e92594eff8d52999b009b33a7
4 N61ed551b82364c9d864ab7ab42474823
5 N6a4668b3ef8f4403aa39aa2e302c0378
6 N932153e875764e8d86e46481e3be052b
7 Nbf8da9ecb5754aae9d292abd805dfe8f
8 Nc62d094b526345238e58534ed9eff303
9 Nd1419adad7204b01a388b6d9598d8f37
10 Nf066e34d78e941a79065f5b5b612b427
11 anzsrc-for:06
12 anzsrc-for:0604
13 schema:author N4cc5bdd69c6a4f649504e5e4186e98a5
14 schema:citation sg:pub.10.1007/s00018-010-0344-4
15 sg:pub.10.1007/s00253-003-1539-5
16 sg:pub.10.1007/s00253-004-1559-9
17 sg:pub.10.1007/s00253-008-1688-7
18 https://doi.org/10.1002/btpr.538
19 https://doi.org/10.1006/prep.2000.1363
20 https://doi.org/10.1016/j.biotechadv.2004.11.003
21 https://doi.org/10.1016/j.enzmictec.2006.10.036
22 https://doi.org/10.1016/j.enzmictec.2008.03.003
23 https://doi.org/10.1016/j.febslet.2008.07.049
24 https://doi.org/10.1016/j.foodchem.2007.04.020
25 https://doi.org/10.1016/j.tibtech.2008.06.006
26 https://doi.org/10.1021/jf703567t
27 https://doi.org/10.1021/jf8008519
28 https://doi.org/10.1046/j.1365-2958.1996.381908.x
29 https://doi.org/10.1101/sqb.1970.035.01.066
30 https://doi.org/10.1111/j.1365-2958.1995.tb02423.x
31 https://doi.org/10.1111/j.1574-6968.2011.02387.x
32 https://doi.org/10.1128/aem.69.1.358-366.2003
33 https://doi.org/10.1128/aem.71.12.8911-8919.2005
34 https://doi.org/10.1177/1087057109356206
35 https://doi.org/10.1271/bbb.58.88
36 https://doi.org/10.1271/bbb.61.830
37 https://doi.org/10.1271/bbb.64.1263
38 https://doi.org/10.1271/bbb.68.2058
39 https://doi.org/10.1271/bbb1961.53.2613
40 https://doi.org/10.5661/bger-26-205
41 schema:datePublished 2011-12
42 schema:datePublishedReg 2011-12-01
43 schema:description BACKGROUND: Streptomyces transglutaminase (TGase) is naturally synthesized as zymogen (pro-TGase), which is then processed to produce active enzyme by the removal of its N-terminal pro-peptide. This pro-peptide is found to be essential for overexpression of soluble TGase in E. coli. However, expression of pro-TGase by E. coli requires protease-mediated activation in vitro. In this study, we developed a novel co- expression method for the direct production of active TGase in E. coli. RESULTS: A TGase from S. hygroscopicus was expressed in E. coli only after fusing with the pelB signal peptide, but fusion with the signal peptide induced insoluble enzyme. Therefore, alternative protocol was designed by co-expressing the TGase and its pro-peptide as independent polypeptides under a single T7 promoter using vector pET-22b(+). Although the pro-peptide was co-expressed, the TGase fused without the signal peptide was undetectable in both soluble and insoluble fractions of the recombinant cells. Similarly, when both genes were expressed in the order of the TGase and the pro-peptide, the solubility of TGase fused with the signal peptide was not improved by the co-expression with its pro-peptide. Interestingly, active TGase was only produced by the cells in which the pro-peptide and the TGase were fused with the signal peptide and sequentially expressed. The purified recombinant and native TGase shared the similar catalytic properties. CONCLUSIONS: Our results indicated that the pro-peptide can assist correct folding of the TGase inter-molecularly in E. coli, and expression of pro-peptide prior to that of TGase was essential for the production of active TGase. The co-expression strategy based on optimizing the order of gene expression could be useful for the expression of other functional proteins that are synthesized as a precursor.
44 schema:genre research_article
45 schema:inLanguage en
46 schema:isAccessibleForFree true
47 schema:isPartOf N1dbdab222802494883ea986bcdce8576
48 Nebb62aad9b254e689238fbd3217de8f5
49 sg:journal.1030599
50 schema:name The order of expression is a key factor in the production of active transglutaminase in Escherichia coli by co-expression with its pro-peptide
51 schema:pagination 112
52 schema:productId N13421501b9814dca83c40226f87829f7
53 N267b30a8f538451bbe84d375bc0c2937
54 N9a1acf3f4e2848ff9eb365ac21077ed8
55 Na2cb306c6f6440299980110993a734e1
56 Ne0d2ff3caf8f43b48780cfa22bf0a678
57 schema:sameAs https://app.dimensions.ai/details/publication/pub.1000744820
58 https://doi.org/10.1186/1475-2859-10-112
59 schema:sdDatePublished 2019-04-10T19:08
60 schema:sdLicense https://scigraph.springernature.com/explorer/license/
61 schema:sdPublisher Nc0d0a575e00a4c229d75d8a8d7daa401
62 schema:url http://link.springer.com/10.1186%2F1475-2859-10-112
63 sgo:license sg:explorer/license/
64 sgo:sdDataset articles
65 rdf:type schema:ScholarlyArticle
66 N128ecf983414492daafd942f8da71df9 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
67 schema:name Molecular Sequence Data
68 rdf:type schema:DefinedTerm
69 N13421501b9814dca83c40226f87829f7 schema:name dimensions_id
70 schema:value pub.1000744820
71 rdf:type schema:PropertyValue
72 N1dbdab222802494883ea986bcdce8576 schema:issueNumber 1
73 rdf:type schema:PublicationIssue
74 N267b30a8f538451bbe84d375bc0c2937 schema:name nlm_unique_id
75 schema:value 101139812
76 rdf:type schema:PropertyValue
77 N41324025c8654ed18c5cd25e9bbc4aa0 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
78 schema:name Escherichia coli
79 rdf:type schema:DefinedTerm
80 N4cc5bdd69c6a4f649504e5e4186e98a5 rdf:first sg:person.0761552653.86
81 rdf:rest Nbf5b0c59f26347d0b1eeaba766590f7d
82 N56f6e09e92594eff8d52999b009b33a7 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
83 schema:name Enzyme Precursors
84 rdf:type schema:DefinedTerm
85 N61ed551b82364c9d864ab7ab42474823 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
86 schema:name Recombinant Fusion Proteins
87 rdf:type schema:DefinedTerm
88 N6379ea31faf94a619816c8eb0ba76fc1 rdf:first sg:person.0774720425.90
89 rdf:rest rdf:nil
90 N6a4668b3ef8f4403aa39aa2e302c0378 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
91 schema:name Protein Sorting Signals
92 rdf:type schema:DefinedTerm
93 N7c262194a67f49f08f669429f41554c4 rdf:first sg:person.01171210206.92
94 rdf:rest N6379ea31faf94a619816c8eb0ba76fc1
95 N932153e875764e8d86e46481e3be052b schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
96 schema:name Transglutaminases
97 rdf:type schema:DefinedTerm
98 N9a1acf3f4e2848ff9eb365ac21077ed8 schema:name pubmed_id
99 schema:value 22196373
100 rdf:type schema:PropertyValue
101 Na2cb306c6f6440299980110993a734e1 schema:name readcube_id
102 schema:value 038c0c43ac4b42f2e7614f17771f3b70e8ac0e650e486329ec02a97384deb4ff
103 rdf:type schema:PropertyValue
104 Nb17964834ca14b488ddd0a42b495e6ab rdf:first sg:person.0775520554.94
105 rdf:rest Nebeb25fd83724d2ea0f3a010a44ac93d
106 Nbe62ac5198114673b4abf11e193864a8 rdf:first sg:person.01303557140.44
107 rdf:rest Nb17964834ca14b488ddd0a42b495e6ab
108 Nbf5b0c59f26347d0b1eeaba766590f7d rdf:first sg:person.01237473145.01
109 rdf:rest Ncca042e1eade4a289c24cdc33876c29c
110 Nbf8da9ecb5754aae9d292abd805dfe8f schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
111 schema:name Streptomyces
112 rdf:type schema:DefinedTerm
113 Nc0d0a575e00a4c229d75d8a8d7daa401 schema:name Springer Nature - SN SciGraph project
114 rdf:type schema:Organization
115 Nc62d094b526345238e58534ed9eff303 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
116 schema:name Bacterial Proteins
117 rdf:type schema:DefinedTerm
118 Ncca042e1eade4a289c24cdc33876c29c rdf:first sg:person.01266055146.55
119 rdf:rest Nbe62ac5198114673b4abf11e193864a8
120 Nd1419adad7204b01a388b6d9598d8f37 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
121 schema:name Protein Transport
122 rdf:type schema:DefinedTerm
123 Ne0d2ff3caf8f43b48780cfa22bf0a678 schema:name doi
124 schema:value 10.1186/1475-2859-10-112
125 rdf:type schema:PropertyValue
126 Nebb62aad9b254e689238fbd3217de8f5 schema:volumeNumber 10
127 rdf:type schema:PublicationVolume
128 Nebeb25fd83724d2ea0f3a010a44ac93d rdf:first sg:person.01263147440.90
129 rdf:rest N7c262194a67f49f08f669429f41554c4
130 Nf066e34d78e941a79065f5b5b612b427 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
131 schema:name Amino Acid Sequence
132 rdf:type schema:DefinedTerm
133 anzsrc-for:06 schema:inDefinedTermSet anzsrc-for:
134 schema:name Biological Sciences
135 rdf:type schema:DefinedTerm
136 anzsrc-for:0604 schema:inDefinedTermSet anzsrc-for:
137 schema:name Genetics
138 rdf:type schema:DefinedTerm
139 sg:grant.4966884 http://pending.schema.org/fundedItem sg:pub.10.1186/1475-2859-10-112
140 rdf:type schema:MonetaryGrant
141 sg:grant.5009460 http://pending.schema.org/fundedItem sg:pub.10.1186/1475-2859-10-112
142 rdf:type schema:MonetaryGrant
143 sg:grant.5014154 http://pending.schema.org/fundedItem sg:pub.10.1186/1475-2859-10-112
144 rdf:type schema:MonetaryGrant
145 sg:grant.7024541 http://pending.schema.org/fundedItem sg:pub.10.1186/1475-2859-10-112
146 rdf:type schema:MonetaryGrant
147 sg:journal.1030599 schema:issn 1475-2859
148 schema:name Microbial Cell Factories
149 rdf:type schema:Periodical
150 sg:person.01171210206.92 schema:affiliation https://www.grid.ac/institutes/grid.258151.a
151 schema:familyName Chen
152 schema:givenName Jian
153 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01171210206.92
154 rdf:type schema:Person
155 sg:person.01237473145.01 schema:affiliation https://www.grid.ac/institutes/grid.258151.a
156 schema:familyName Zhang
157 schema:givenName Dongxu
158 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01237473145.01
159 rdf:type schema:Person
160 sg:person.01263147440.90 schema:affiliation https://www.grid.ac/institutes/grid.258151.a
161 schema:familyName Du
162 schema:givenName Guocheng
163 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01263147440.90
164 rdf:type schema:Person
165 sg:person.01266055146.55 schema:affiliation https://www.grid.ac/institutes/grid.258151.a
166 schema:familyName Wang
167 schema:givenName Miao
168 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01266055146.55
169 rdf:type schema:Person
170 sg:person.01303557140.44 schema:affiliation https://www.grid.ac/institutes/grid.258151.a
171 schema:familyName Cui
172 schema:givenName Wenjing
173 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01303557140.44
174 rdf:type schema:Person
175 sg:person.0761552653.86 schema:affiliation https://www.grid.ac/institutes/grid.258151.a
176 schema:familyName Liu
177 schema:givenName Song
178 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0761552653.86
179 rdf:type schema:Person
180 sg:person.0774720425.90 schema:affiliation https://www.grid.ac/institutes/grid.258151.a
181 schema:familyName Zhou
182 schema:givenName Zhemin
183 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0774720425.90
184 rdf:type schema:Person
185 sg:person.0775520554.94 schema:affiliation https://www.grid.ac/institutes/grid.258151.a
186 schema:familyName Chen
187 schema:givenName Kangkang
188 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0775520554.94
189 rdf:type schema:Person
190 sg:pub.10.1007/s00018-010-0344-4 schema:sameAs https://app.dimensions.ai/details/publication/pub.1027832987
191 https://doi.org/10.1007/s00018-010-0344-4
192 rdf:type schema:CreativeWork
193 sg:pub.10.1007/s00253-003-1539-5 schema:sameAs https://app.dimensions.ai/details/publication/pub.1040145592
194 https://doi.org/10.1007/s00253-003-1539-5
195 rdf:type schema:CreativeWork
196 sg:pub.10.1007/s00253-004-1559-9 schema:sameAs https://app.dimensions.ai/details/publication/pub.1009268886
197 https://doi.org/10.1007/s00253-004-1559-9
198 rdf:type schema:CreativeWork
199 sg:pub.10.1007/s00253-008-1688-7 schema:sameAs https://app.dimensions.ai/details/publication/pub.1011652088
200 https://doi.org/10.1007/s00253-008-1688-7
201 rdf:type schema:CreativeWork
202 https://doi.org/10.1002/btpr.538 schema:sameAs https://app.dimensions.ai/details/publication/pub.1034990825
203 rdf:type schema:CreativeWork
204 https://doi.org/10.1006/prep.2000.1363 schema:sameAs https://app.dimensions.ai/details/publication/pub.1010297488
205 rdf:type schema:CreativeWork
206 https://doi.org/10.1016/j.biotechadv.2004.11.003 schema:sameAs https://app.dimensions.ai/details/publication/pub.1020388916
207 rdf:type schema:CreativeWork
208 https://doi.org/10.1016/j.enzmictec.2006.10.036 schema:sameAs https://app.dimensions.ai/details/publication/pub.1050088038
209 rdf:type schema:CreativeWork
210 https://doi.org/10.1016/j.enzmictec.2008.03.003 schema:sameAs https://app.dimensions.ai/details/publication/pub.1022019325
211 rdf:type schema:CreativeWork
212 https://doi.org/10.1016/j.febslet.2008.07.049 schema:sameAs https://app.dimensions.ai/details/publication/pub.1031701399
213 rdf:type schema:CreativeWork
214 https://doi.org/10.1016/j.foodchem.2007.04.020 schema:sameAs https://app.dimensions.ai/details/publication/pub.1046226945
215 rdf:type schema:CreativeWork
216 https://doi.org/10.1016/j.tibtech.2008.06.006 schema:sameAs https://app.dimensions.ai/details/publication/pub.1034786742
217 rdf:type schema:CreativeWork
218 https://doi.org/10.1021/jf703567t schema:sameAs https://app.dimensions.ai/details/publication/pub.1055924478
219 rdf:type schema:CreativeWork
220 https://doi.org/10.1021/jf8008519 schema:sameAs https://app.dimensions.ai/details/publication/pub.1055924931
221 rdf:type schema:CreativeWork
222 https://doi.org/10.1046/j.1365-2958.1996.381908.x schema:sameAs https://app.dimensions.ai/details/publication/pub.1023177545
223 rdf:type schema:CreativeWork
224 https://doi.org/10.1101/sqb.1970.035.01.066 schema:sameAs https://app.dimensions.ai/details/publication/pub.1060404102
225 rdf:type schema:CreativeWork
226 https://doi.org/10.1111/j.1365-2958.1995.tb02423.x schema:sameAs https://app.dimensions.ai/details/publication/pub.1050201312
227 rdf:type schema:CreativeWork
228 https://doi.org/10.1111/j.1574-6968.2011.02387.x schema:sameAs https://app.dimensions.ai/details/publication/pub.1024207652
229 rdf:type schema:CreativeWork
230 https://doi.org/10.1128/aem.69.1.358-366.2003 schema:sameAs https://app.dimensions.ai/details/publication/pub.1033892926
231 rdf:type schema:CreativeWork
232 https://doi.org/10.1128/aem.71.12.8911-8919.2005 schema:sameAs https://app.dimensions.ai/details/publication/pub.1017702218
233 rdf:type schema:CreativeWork
234 https://doi.org/10.1177/1087057109356206 schema:sameAs https://app.dimensions.ai/details/publication/pub.1063973629
235 rdf:type schema:CreativeWork
236 https://doi.org/10.1271/bbb.58.88 schema:sameAs https://app.dimensions.ai/details/publication/pub.1005551430
237 rdf:type schema:CreativeWork
238 https://doi.org/10.1271/bbb.61.830 schema:sameAs https://app.dimensions.ai/details/publication/pub.1046482878
239 rdf:type schema:CreativeWork
240 https://doi.org/10.1271/bbb.64.1263 schema:sameAs https://app.dimensions.ai/details/publication/pub.1021253186
241 rdf:type schema:CreativeWork
242 https://doi.org/10.1271/bbb.68.2058 schema:sameAs https://app.dimensions.ai/details/publication/pub.1021692720
243 rdf:type schema:CreativeWork
244 https://doi.org/10.1271/bbb1961.53.2613 schema:sameAs https://app.dimensions.ai/details/publication/pub.1007986290
245 rdf:type schema:CreativeWork
246 https://doi.org/10.5661/bger-26-205 schema:sameAs https://app.dimensions.ai/details/publication/pub.1022425337
247 rdf:type schema:CreativeWork
248 https://www.grid.ac/institutes/grid.258151.a schema:alternateName Jiangnan University
249 schema:name School of Biotechnology, Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Lihu Avenue, Wuxi, China
250 School of Food Science and Technology, Jiangnan University, Lihu Avenue, Wuxi, China
251 State Key Laboratory of Food Science and Technology, Jiangnan University, Lihu Avenue, Wuxi, China
252 rdf:type schema:Organization
 




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


...