Engineering a Pichia pastoris nitrilase whole cell catalyst through the increased nitrilase gene copy number and co-expressing of ER oxidoreductin ... View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2020-02-04

AUTHORS

Qi Shen, Zhuang Yu, Pei-jin Lv, Qian Li, Shu-Ping Zou, Neng Xiong, Zhi-Qiang Liu, Ya-Ping Xue, Yu-Guo Zheng

ABSTRACT

1-Cyanocyclohexaneacetic acid (1-CHAA) is a critical intermediate for the synthesis of the antiepileptic agent gabapentin. Previously, our group has established a novel manufacturing route for 1-CHAA through bioconversion catalyzed by an Escherichia coli (E. coli) nitrilase whole cell catalyst. However, the nitrilase expressed in E. coli has several drawbacks such as a low level of reusability, which hampered its industrial application. Herein, we investigated the potential of using the methylotrophic yeast Pichia pastoris (P. pastoris) for producing the nitrilase whole cell catalyst. To achieve strains with high catalytic activities, we investigated the effects of the promoter choice, expressing cassette copy number, and co-expression of chaperone on the production of nitrilase. Our results demonstrated that the strain harboring the multicopy integrations of nitrilase gene under the control of the alcohol oxidase 1 (AOX1) promoter and co-expressing of ER oxidoreductin 1 (ERO1) exhibited an 18-fold enhancement in the nitrilase activity compared with the strain containing a single integration of nitrilase gene under the control of glyceraldehyde-3-phosphate (GAP) dehydrogenase promoter. This optimized P. pastoris strain, compared with the E. coli nitrilase whole cell catalyst, shows greatly improved levels of reusability and thermostability while has a similar high-substrate tolerance. More... »

PAGES

2489-2500

References to SciGraph publications

  • 2011-09-03. Purification and characterization of heterologously expressed nitrilases from filamentous fungi in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2014-01-04. Engineering of a Pichia pastoris Expression System for High-Level Secretion of HSA/GH Fusion Protein in APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
  • 2013-08-30. Efficient secretion of lipase r27RCL in Pichia pastoris by enhancing the disulfide bond formation pathway in the endoplasmic reticulum in JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
  • 2018-11-28. Highly efficient conversion of 1-cyanocycloalkaneacetonitrile using a “super nitrilase mutant” in BIOPROCESS AND BIOSYSTEMS ENGINEERING
  • 2011-05-10. Increasing gene dosage greatly enhances recombinant expression of aquaporins in Pichia pastoris in BMC BIOTECHNOLOGY
  • 2014-02-13. 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
  • 2012-08-12. The effect of gene copy number and co-expression of chaperone on production of albumin fusion proteins in Pichia pastoris in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2012-08-08. 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
  • 2014-04-18. Protein expression in Pichia pastoris: recent achievements and perspectives for heterologous protein production in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2016-11-17. Recent advances and challenges in the heterologous production of microbial nitrilases for biocatalytic applications in WORLD JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
  • 2013-12-20. Can too many copies spoil the broth? in MICROBIAL CELL FACTORIES
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/s00253-020-10422-4

    DOI

    http://dx.doi.org/10.1007/s00253-020-10422-4

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1124590256

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/32020278


    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/2008/0604", 
            "inDefinedTermSet": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/", 
            "name": "Genetics", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "Aminohydrolases", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "Catalysis", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "Gene Dosage", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "Oxidoreductases Acting on Sulfur Group Donors", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "Pichia", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "Promoter Regions, Genetic", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "Protein Engineering", 
            "type": "DefinedTerm"
          }
        ], 
        "author": [
          {
            "affiliation": {
              "alternateName": "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
              "id": "http://www.grid.ac/institutes/grid.469325.f", 
              "name": [
                "Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
                "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Shen", 
            "givenName": "Qi", 
            "id": "sg:person.014021504107.94", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.014021504107.94"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
              "id": "http://www.grid.ac/institutes/grid.469325.f", 
              "name": [
                "Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
                "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Yu", 
            "givenName": "Zhuang", 
            "id": "sg:person.011244547115.09", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.011244547115.09"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
              "id": "http://www.grid.ac/institutes/grid.469325.f", 
              "name": [
                "Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
                "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Lv", 
            "givenName": "Pei-jin", 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
              "id": "http://www.grid.ac/institutes/grid.469325.f", 
              "name": [
                "Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
                "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Li", 
            "givenName": "Qian", 
            "id": "sg:person.016422772515.43", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.016422772515.43"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
              "id": "http://www.grid.ac/institutes/grid.469325.f", 
              "name": [
                "Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
                "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Zou", 
            "givenName": "Shu-Ping", 
            "id": "sg:person.01156710533.23", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01156710533.23"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
              "id": "http://www.grid.ac/institutes/grid.469325.f", 
              "name": [
                "Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
                "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Xiong", 
            "givenName": "Neng", 
            "id": "sg:person.014740123500.86", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.014740123500.86"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
              "id": "http://www.grid.ac/institutes/grid.469325.f", 
              "name": [
                "Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
                "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Liu", 
            "givenName": "Zhi-Qiang", 
            "id": "sg:person.01256734541.85", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01256734541.85"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
              "id": "http://www.grid.ac/institutes/grid.469325.f", 
              "name": [
                "Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
                "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Xue", 
            "givenName": "Ya-Ping", 
            "id": "sg:person.0744724672.04", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0744724672.04"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
              "id": "http://www.grid.ac/institutes/grid.469325.f", 
              "name": [
                "Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China", 
                "Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People\u2019s Republic of China"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Zheng", 
            "givenName": "Yu-Guo", 
            "id": "sg:person.01144225405.06", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01144225405.06"
            ], 
            "type": "Person"
          }
        ], 
        "citation": [
          {
            "id": "sg:pub.10.1186/1475-2859-12-128", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1048533634", 
              "https://doi.org/10.1186/1475-2859-12-128"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s00253-012-4337-0", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1031502416", 
              "https://doi.org/10.1007/s00253-012-4337-0"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s11274-016-2173-6", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1021710199", 
              "https://doi.org/10.1007/s11274-016-2173-6"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s00253-014-5732-5", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1027836620", 
              "https://doi.org/10.1007/s00253-014-5732-5"
            ], 
            "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.10.1007/s00449-018-2049-2", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1110263506", 
              "https://doi.org/10.1007/s00449-018-2049-2"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s12010-013-0688-y", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1050438898", 
              "https://doi.org/10.1007/s12010-013-0688-y"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s10295-013-1328-9", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1039166515", 
              "https://doi.org/10.1007/s10295-013-1328-9"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s00253-011-3525-7", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1001069513", 
              "https://doi.org/10.1007/s00253-011-3525-7"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1186/1472-6750-11-47", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1028680395", 
              "https://doi.org/10.1186/1472-6750-11-47"
            ], 
            "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"
          }
        ], 
        "datePublished": "2020-02-04", 
        "datePublishedReg": "2020-02-04", 
        "description": "Abstract1-Cyanocyclohexaneacetic acid (1-CHAA) is a critical intermediate for the synthesis of the antiepileptic agent gabapentin. Previously, our group has established a novel manufacturing route for 1-CHAA through bioconversion catalyzed by an Escherichia coli (E. coli) nitrilase whole cell catalyst. However, the nitrilase expressed in E. coli has several drawbacks such as a low level of reusability, which hampered its industrial application. Herein, we investigated the potential of using the methylotrophic yeast Pichia pastoris (P. pastoris) for producing the nitrilase whole cell catalyst. To achieve strains with high catalytic activities, we investigated the effects of the promoter choice, expressing cassette copy number, and co-expression of chaperone on the production of nitrilase. Our results demonstrated that the strain harboring the multicopy integrations of nitrilase gene under the control of the alcohol oxidase 1 (AOX1) promoter and co-expressing of ER oxidoreductin 1 (ERO1) exhibited an 18-fold enhancement in the nitrilase activity compared with the strain containing a single integration of nitrilase gene under the control of glyceraldehyde-3-phosphate (GAP) dehydrogenase promoter. This optimized P. pastoris strain, compared with the E. coli nitrilase whole cell catalyst, shows greatly improved levels of reusability and thermostability while has a similar high-substrate tolerance.", 
        "genre": "article", 
        "id": "sg:pub.10.1007/s00253-020-10422-4", 
        "inLanguage": "en", 
        "isAccessibleForFree": false, 
        "isPartOf": [
          {
            "id": "sg:journal.1083533", 
            "issn": [
              "0175-7598", 
              "1432-0614"
            ], 
            "name": "Applied Microbiology and Biotechnology", 
            "publisher": "Springer Nature", 
            "type": "Periodical"
          }, 
          {
            "issueNumber": "6", 
            "type": "PublicationIssue"
          }, 
          {
            "type": "PublicationVolume", 
            "volumeNumber": "104"
          }
        ], 
        "keywords": [
          "whole-cell catalyst", 
          "cell catalysts", 
          "nitrilase gene", 
          "Pichia pastoris", 
          "methylotrophic yeast Pichia pastoris", 
          "alcohol oxidase 1 promoter", 
          "yeast Pichia pastoris", 
          "high catalytic activity", 
          "glyceraldehyde-3-phosphate dehydrogenase promoter", 
          "multicopy integration", 
          "pastoris strain", 
          "high substrate tolerance", 
          "catalytic activity", 
          "promoter choice", 
          "nitrilase activity", 
          "dehydrogenase promoter", 
          "industrial applications", 
          "catalyst", 
          "pastoris", 
          "nitrilase", 
          "reusability", 
          "novel manufacturing route", 
          "copy number", 
          "single integration", 
          "gene copy number", 
          "bioconversion", 
          "manufacturing route", 
          "Herein", 
          "Escherichia coli", 
          "integration", 
          "coli", 
          "thermostability", 
          "route", 
          "applications", 
          "synthesis", 
          "drawbacks", 
          "enhancement", 
          "strains", 
          "promoter", 
          "potential", 
          "production", 
          "ER oxidoreductin 1", 
          "acid", 
          "genes", 
          "activity", 
          "critical intermediate", 
          "control", 
          "tolerance", 
          "number", 
          "results", 
          "improved levels", 
          "intermediates", 
          "low levels", 
          "effect", 
          "levels", 
          "choice", 
          "chaperones", 
          "antiepileptic agent gabapentin", 
          "group", 
          "gabapentin", 
          "Abstract1-Cyanocyclohexaneacetic acid", 
          "agent gabapentin", 
          "nitrilase whole cell catalyst", 
          "cassette copy number", 
          "production of nitrilase", 
          "oxidase 1 (AOX1) promoter", 
          "oxidoreductin 1", 
          "coli nitrilase whole cell catalyst", 
          "similar high-substrate tolerance", 
          "nitrilase gene copy number"
        ], 
        "name": "Engineering a Pichia pastoris nitrilase whole cell catalyst through the increased nitrilase gene copy number and co-expressing of ER oxidoreductin 1", 
        "pagination": "2489-2500", 
        "productId": [
          {
            "name": "dimensions_id", 
            "type": "PropertyValue", 
            "value": [
              "pub.1124590256"
            ]
          }, 
          {
            "name": "doi", 
            "type": "PropertyValue", 
            "value": [
              "10.1007/s00253-020-10422-4"
            ]
          }, 
          {
            "name": "pubmed_id", 
            "type": "PropertyValue", 
            "value": [
              "32020278"
            ]
          }
        ], 
        "sameAs": [
          "https://doi.org/10.1007/s00253-020-10422-4", 
          "https://app.dimensions.ai/details/publication/pub.1124590256"
        ], 
        "sdDataset": "articles", 
        "sdDatePublished": "2022-01-01T18:56", 
        "sdLicense": "https://scigraph.springernature.com/explorer/license/", 
        "sdPublisher": {
          "name": "Springer Nature - SN SciGraph project", 
          "type": "Organization"
        }, 
        "sdSource": "s3://com-springernature-scigraph/baseset/20220101/entities/gbq_results/article/article_851.jsonl", 
        "type": "ScholarlyArticle", 
        "url": "https://doi.org/10.1007/s00253-020-10422-4"
      }
    ]
     

    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.10.1007/s00253-020-10422-4'

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

    curl -H 'Accept: application/n-triples' 'https://scigraph.springernature.com/pub.10.1007/s00253-020-10422-4'

    Turtle is a human-readable linked data format.

    curl -H 'Accept: text/turtle' 'https://scigraph.springernature.com/pub.10.1007/s00253-020-10422-4'

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

    curl -H 'Accept: application/rdf+xml' 'https://scigraph.springernature.com/pub.10.1007/s00253-020-10422-4'


     

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

    260 TRIPLES      22 PREDICATES      114 URIs      95 LITERALS      14 BLANK NODES

    Subject Predicate Object
    1 sg:pub.10.1007/s00253-020-10422-4 schema:about N3a4f060ef5e74d5e9e59a6e95e1d312f
    2 N6fdaac820d724726983de39a2270bf94
    3 N86fc3395fcf343f39e5c554b52424e54
    4 N8d1c586dae75497fa3c5d82f18551efa
    5 N94475116e1fd4e46af3fd4a7f27a43d6
    6 Nd3116cbba94d492d87c3a84d5152cac0
    7 Ndfcb1ad648d6402da0841a3f9993e6fb
    8 anzsrc-for:06
    9 anzsrc-for:0604
    10 schema:author N29b0d890149e4921a33f1a62d4712a6d
    11 schema:citation sg:pub.10.1007/s00253-011-3525-7
    12 sg:pub.10.1007/s00253-012-4337-0
    13 sg:pub.10.1007/s00253-014-5732-5
    14 sg:pub.10.1007/s00449-018-2049-2
    15 sg:pub.10.1007/s10295-013-1328-9
    16 sg:pub.10.1007/s11274-016-2173-6
    17 sg:pub.10.1007/s12010-013-0688-y
    18 sg:pub.10.1186/1472-6750-11-47
    19 sg:pub.10.1186/1475-2859-11-103
    20 sg:pub.10.1186/1475-2859-12-128
    21 sg:pub.10.1186/1475-2859-13-23
    22 schema:datePublished 2020-02-04
    23 schema:datePublishedReg 2020-02-04
    24 schema:description Abstract1-Cyanocyclohexaneacetic acid (1-CHAA) is a critical intermediate for the synthesis of the antiepileptic agent gabapentin. Previously, our group has established a novel manufacturing route for 1-CHAA through bioconversion catalyzed by an Escherichia coli (E. coli) nitrilase whole cell catalyst. However, the nitrilase expressed in E. coli has several drawbacks such as a low level of reusability, which hampered its industrial application. Herein, we investigated the potential of using the methylotrophic yeast Pichia pastoris (P. pastoris) for producing the nitrilase whole cell catalyst. To achieve strains with high catalytic activities, we investigated the effects of the promoter choice, expressing cassette copy number, and co-expression of chaperone on the production of nitrilase. Our results demonstrated that the strain harboring the multicopy integrations of nitrilase gene under the control of the alcohol oxidase 1 (AOX1) promoter and co-expressing of ER oxidoreductin 1 (ERO1) exhibited an 18-fold enhancement in the nitrilase activity compared with the strain containing a single integration of nitrilase gene under the control of glyceraldehyde-3-phosphate (GAP) dehydrogenase promoter. This optimized P. pastoris strain, compared with the E. coli nitrilase whole cell catalyst, shows greatly improved levels of reusability and thermostability while has a similar high-substrate tolerance.
    25 schema:genre article
    26 schema:inLanguage en
    27 schema:isAccessibleForFree false
    28 schema:isPartOf N557d13ffb84440eabe60ba13b12c35b7
    29 Nb9a1ff41e24e4feea6c2ce96f10a923f
    30 sg:journal.1083533
    31 schema:keywords Abstract1-Cyanocyclohexaneacetic acid
    32 ER oxidoreductin 1
    33 Escherichia coli
    34 Herein
    35 Pichia pastoris
    36 acid
    37 activity
    38 agent gabapentin
    39 alcohol oxidase 1 promoter
    40 antiepileptic agent gabapentin
    41 applications
    42 bioconversion
    43 cassette copy number
    44 catalyst
    45 catalytic activity
    46 cell catalysts
    47 chaperones
    48 choice
    49 coli
    50 coli nitrilase whole cell catalyst
    51 control
    52 copy number
    53 critical intermediate
    54 dehydrogenase promoter
    55 drawbacks
    56 effect
    57 enhancement
    58 gabapentin
    59 gene copy number
    60 genes
    61 glyceraldehyde-3-phosphate dehydrogenase promoter
    62 group
    63 high catalytic activity
    64 high substrate tolerance
    65 improved levels
    66 industrial applications
    67 integration
    68 intermediates
    69 levels
    70 low levels
    71 manufacturing route
    72 methylotrophic yeast Pichia pastoris
    73 multicopy integration
    74 nitrilase
    75 nitrilase activity
    76 nitrilase gene
    77 nitrilase gene copy number
    78 nitrilase whole cell catalyst
    79 novel manufacturing route
    80 number
    81 oxidase 1 (AOX1) promoter
    82 oxidoreductin 1
    83 pastoris
    84 pastoris strain
    85 potential
    86 production
    87 production of nitrilase
    88 promoter
    89 promoter choice
    90 results
    91 reusability
    92 route
    93 similar high-substrate tolerance
    94 single integration
    95 strains
    96 synthesis
    97 thermostability
    98 tolerance
    99 whole-cell catalyst
    100 yeast Pichia pastoris
    101 schema:name Engineering a Pichia pastoris nitrilase whole cell catalyst through the increased nitrilase gene copy number and co-expressing of ER oxidoreductin 1
    102 schema:pagination 2489-2500
    103 schema:productId N399ed13a5d7146128e3d6febeea669ea
    104 N608d403e4f20481493618eb098c8bde6
    105 Naf2989fa7916481fb160bdd49c57afb7
    106 schema:sameAs https://app.dimensions.ai/details/publication/pub.1124590256
    107 https://doi.org/10.1007/s00253-020-10422-4
    108 schema:sdDatePublished 2022-01-01T18:56
    109 schema:sdLicense https://scigraph.springernature.com/explorer/license/
    110 schema:sdPublisher Nce51e4ad289f4b46be2314fc58c399d3
    111 schema:url https://doi.org/10.1007/s00253-020-10422-4
    112 sgo:license sg:explorer/license/
    113 sgo:sdDataset articles
    114 rdf:type schema:ScholarlyArticle
    115 N127b7bc94a3445c09e20ae12c8bde1b0 rdf:first N181f211689e14509825d756abcda4c17
    116 rdf:rest N7419aa2c4f834bcea3fc15f7e6229698
    117 N181f211689e14509825d756abcda4c17 schema:affiliation grid-institutes:grid.469325.f
    118 schema:familyName Lv
    119 schema:givenName Pei-jin
    120 rdf:type schema:Person
    121 N1a5379b78f6b44ceb824acd81e50ac3d rdf:first sg:person.011244547115.09
    122 rdf:rest N127b7bc94a3445c09e20ae12c8bde1b0
    123 N29b0d890149e4921a33f1a62d4712a6d rdf:first sg:person.014021504107.94
    124 rdf:rest N1a5379b78f6b44ceb824acd81e50ac3d
    125 N399ed13a5d7146128e3d6febeea669ea schema:name pubmed_id
    126 schema:value 32020278
    127 rdf:type schema:PropertyValue
    128 N3a4f060ef5e74d5e9e59a6e95e1d312f schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    129 schema:name Oxidoreductases Acting on Sulfur Group Donors
    130 rdf:type schema:DefinedTerm
    131 N3a6476455a2a446e96a7381f011a88bf rdf:first sg:person.01256734541.85
    132 rdf:rest N59666d97936444dd8d89dbcd0fbc3783
    133 N557d13ffb84440eabe60ba13b12c35b7 schema:issueNumber 6
    134 rdf:type schema:PublicationIssue
    135 N56011677cd7342e6866cd214f0a03861 rdf:first sg:person.01156710533.23
    136 rdf:rest N8e025a0a9af54e18957995ed4df9190c
    137 N59666d97936444dd8d89dbcd0fbc3783 rdf:first sg:person.0744724672.04
    138 rdf:rest N9cfadcd23cba48a09c38dc0aa3c1ba3e
    139 N608d403e4f20481493618eb098c8bde6 schema:name dimensions_id
    140 schema:value pub.1124590256
    141 rdf:type schema:PropertyValue
    142 N6fdaac820d724726983de39a2270bf94 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    143 schema:name Gene Dosage
    144 rdf:type schema:DefinedTerm
    145 N7419aa2c4f834bcea3fc15f7e6229698 rdf:first sg:person.016422772515.43
    146 rdf:rest N56011677cd7342e6866cd214f0a03861
    147 N86fc3395fcf343f39e5c554b52424e54 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    148 schema:name Protein Engineering
    149 rdf:type schema:DefinedTerm
    150 N8d1c586dae75497fa3c5d82f18551efa schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    151 schema:name Promoter Regions, Genetic
    152 rdf:type schema:DefinedTerm
    153 N8e025a0a9af54e18957995ed4df9190c rdf:first sg:person.014740123500.86
    154 rdf:rest N3a6476455a2a446e96a7381f011a88bf
    155 N94475116e1fd4e46af3fd4a7f27a43d6 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    156 schema:name Aminohydrolases
    157 rdf:type schema:DefinedTerm
    158 N9cfadcd23cba48a09c38dc0aa3c1ba3e rdf:first sg:person.01144225405.06
    159 rdf:rest rdf:nil
    160 Naf2989fa7916481fb160bdd49c57afb7 schema:name doi
    161 schema:value 10.1007/s00253-020-10422-4
    162 rdf:type schema:PropertyValue
    163 Nb9a1ff41e24e4feea6c2ce96f10a923f schema:volumeNumber 104
    164 rdf:type schema:PublicationVolume
    165 Nce51e4ad289f4b46be2314fc58c399d3 schema:name Springer Nature - SN SciGraph project
    166 rdf:type schema:Organization
    167 Nd3116cbba94d492d87c3a84d5152cac0 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    168 schema:name Pichia
    169 rdf:type schema:DefinedTerm
    170 Ndfcb1ad648d6402da0841a3f9993e6fb schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    171 schema:name Catalysis
    172 rdf:type schema:DefinedTerm
    173 anzsrc-for:06 schema:inDefinedTermSet anzsrc-for:
    174 schema:name Biological Sciences
    175 rdf:type schema:DefinedTerm
    176 anzsrc-for:0604 schema:inDefinedTermSet anzsrc-for:
    177 schema:name Genetics
    178 rdf:type schema:DefinedTerm
    179 sg:journal.1083533 schema:issn 0175-7598
    180 1432-0614
    181 schema:name Applied Microbiology and Biotechnology
    182 schema:publisher Springer Nature
    183 rdf:type schema:Periodical
    184 sg:person.011244547115.09 schema:affiliation grid-institutes:grid.469325.f
    185 schema:familyName Yu
    186 schema:givenName Zhuang
    187 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.011244547115.09
    188 rdf:type schema:Person
    189 sg:person.01144225405.06 schema:affiliation grid-institutes:grid.469325.f
    190 schema:familyName Zheng
    191 schema:givenName Yu-Guo
    192 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01144225405.06
    193 rdf:type schema:Person
    194 sg:person.01156710533.23 schema:affiliation grid-institutes:grid.469325.f
    195 schema:familyName Zou
    196 schema:givenName Shu-Ping
    197 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01156710533.23
    198 rdf:type schema:Person
    199 sg:person.01256734541.85 schema:affiliation grid-institutes:grid.469325.f
    200 schema:familyName Liu
    201 schema:givenName Zhi-Qiang
    202 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01256734541.85
    203 rdf:type schema:Person
    204 sg:person.014021504107.94 schema:affiliation grid-institutes:grid.469325.f
    205 schema:familyName Shen
    206 schema:givenName Qi
    207 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.014021504107.94
    208 rdf:type schema:Person
    209 sg:person.014740123500.86 schema:affiliation grid-institutes:grid.469325.f
    210 schema:familyName Xiong
    211 schema:givenName Neng
    212 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.014740123500.86
    213 rdf:type schema:Person
    214 sg:person.016422772515.43 schema:affiliation grid-institutes:grid.469325.f
    215 schema:familyName Li
    216 schema:givenName Qian
    217 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.016422772515.43
    218 rdf:type schema:Person
    219 sg:person.0744724672.04 schema:affiliation grid-institutes:grid.469325.f
    220 schema:familyName Xue
    221 schema:givenName Ya-Ping
    222 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0744724672.04
    223 rdf:type schema:Person
    224 sg:pub.10.1007/s00253-011-3525-7 schema:sameAs https://app.dimensions.ai/details/publication/pub.1001069513
    225 https://doi.org/10.1007/s00253-011-3525-7
    226 rdf:type schema:CreativeWork
    227 sg:pub.10.1007/s00253-012-4337-0 schema:sameAs https://app.dimensions.ai/details/publication/pub.1031502416
    228 https://doi.org/10.1007/s00253-012-4337-0
    229 rdf:type schema:CreativeWork
    230 sg:pub.10.1007/s00253-014-5732-5 schema:sameAs https://app.dimensions.ai/details/publication/pub.1027836620
    231 https://doi.org/10.1007/s00253-014-5732-5
    232 rdf:type schema:CreativeWork
    233 sg:pub.10.1007/s00449-018-2049-2 schema:sameAs https://app.dimensions.ai/details/publication/pub.1110263506
    234 https://doi.org/10.1007/s00449-018-2049-2
    235 rdf:type schema:CreativeWork
    236 sg:pub.10.1007/s10295-013-1328-9 schema:sameAs https://app.dimensions.ai/details/publication/pub.1039166515
    237 https://doi.org/10.1007/s10295-013-1328-9
    238 rdf:type schema:CreativeWork
    239 sg:pub.10.1007/s11274-016-2173-6 schema:sameAs https://app.dimensions.ai/details/publication/pub.1021710199
    240 https://doi.org/10.1007/s11274-016-2173-6
    241 rdf:type schema:CreativeWork
    242 sg:pub.10.1007/s12010-013-0688-y schema:sameAs https://app.dimensions.ai/details/publication/pub.1050438898
    243 https://doi.org/10.1007/s12010-013-0688-y
    244 rdf:type schema:CreativeWork
    245 sg:pub.10.1186/1472-6750-11-47 schema:sameAs https://app.dimensions.ai/details/publication/pub.1028680395
    246 https://doi.org/10.1186/1472-6750-11-47
    247 rdf:type schema:CreativeWork
    248 sg:pub.10.1186/1475-2859-11-103 schema:sameAs https://app.dimensions.ai/details/publication/pub.1051848306
    249 https://doi.org/10.1186/1475-2859-11-103
    250 rdf:type schema:CreativeWork
    251 sg:pub.10.1186/1475-2859-12-128 schema:sameAs https://app.dimensions.ai/details/publication/pub.1048533634
    252 https://doi.org/10.1186/1475-2859-12-128
    253 rdf:type schema:CreativeWork
    254 sg:pub.10.1186/1475-2859-13-23 schema:sameAs https://app.dimensions.ai/details/publication/pub.1051324720
    255 https://doi.org/10.1186/1475-2859-13-23
    256 rdf:type schema:CreativeWork
    257 grid-institutes:grid.469325.f schema:alternateName Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People’s Republic of China
    258 schema:name Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, 310014, Hangzhou, People’s Republic of China
    259 Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, People’s Republic of China
    260 rdf:type schema:Organization
     




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


    ...