Mutational analysis of the hyc-operon determining the relationship between hydrogenase-3 and NADH pathway in Enterobacter aerogenes.

Saved in:
Bibliographic Details
Title: Mutational analysis of the hyc-operon determining the relationship between hydrogenase-3 and NADH pathway in Enterobacter aerogenes.
Authors: Pi, Jian1,2, Jawed, Muhammad1,2, Wang, Jun1,2, Xu, Li1,2 xuli@hust.edu.cn, Yan, Yunjun1,2 yanyunjun@hust.edu.cn
Source: Enzyme & Microbial Technology. Jan2016, Vol. 82, p1-7. 7p.
Subjects: Operons, Hydrogenase, Genetic mutation, NAD (Coenzyme), Enterobacter aerogenes, Hydrogen production
Abstract: In this study, the hydrogenase-3 gene cluster ( hycDEFGH ) was isolated and identified from Enterobacter aerogenes CCTCC AB91102. All gene products were highly homologous to the reported bacterial hydrogenase-3 (Hyd-3) proteins. The genes hycE , hycF , hycG encoding the subunits of hydrogenase-3 were targeted for genetic knockout to inhibit the FHL hydrogen production pathway via the Red recombination system, generating three mutant strains AB91102-E (Δ hycE ), AB91102-F (Δ hycF ) and AB91102-G (Δ hycG ). Deletion of the three genes affected the integrity of hydrogenase-3. The hydrogen production experiments with the mutant strains showed that no hydrogen was detected compared with the wild type (0.886 mol/mol glucose), demonstrating that knocking out any of the three genes could inhibit NADH hydrogen production pathway. Meanwhile, the metabolites of the mutant strains were significantly changed in comparison with the wild type, indicating corresponding changes in metabolic flux by mutation. Additionally, the activity of NADH-mediated hydrogenase was found to be nil in the mutant strains. The chemostat experiments showed that the NADH/NAD + ratio of the mutant strains increased nearly 1.4-fold compared with the wild type. The NADH-mediated hydrogenase activity and NADH/NAD + ratio analysis both suggested that NADH pathway required the involvement of the electron transport chain of hydrogenase-3. [ABSTRACT FROM AUTHOR]
Copyright of Enzyme & Microbial Technology is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
Database: Engineering Source
FullText Text:
  Availability: 0
Header DbId: egs
DbLabel: Engineering Source
An: 111498234
AccessLevel: 6
PubType: Academic Journal
PubTypeId: academicJournal
PreciseRelevancyScore: 0
IllustrationInfo
Items – Name: Title
  Label: Title
  Group: Ti
  Data: Mutational analysis of the hyc-operon determining the relationship between hydrogenase-3 and NADH pathway in Enterobacter aerogenes.
– Name: Author
  Label: Authors
  Group: Au
  Data: <searchLink fieldCode="AR" term="%22Pi%2C+Jian%22">Pi, Jian</searchLink><relatesTo>1,2</relatesTo><br /><searchLink fieldCode="AR" term="%22Jawed%2C+Muhammad%22">Jawed, Muhammad</searchLink><relatesTo>1,2</relatesTo><br /><searchLink fieldCode="AR" term="%22Wang%2C+Jun%22">Wang, Jun</searchLink><relatesTo>1,2</relatesTo><br /><searchLink fieldCode="AR" term="%22Xu%2C+Li%22">Xu, Li</searchLink><relatesTo>1,2</relatesTo><i> xuli@hust.edu.cn</i><br /><searchLink fieldCode="AR" term="%22Yan%2C+Yunjun%22">Yan, Yunjun</searchLink><relatesTo>1,2</relatesTo><i> yanyunjun@hust.edu.cn</i>
– Name: TitleSource
  Label: Source
  Group: Src
  Data: <searchLink fieldCode="JN" term="%22Enzyme+%26+Microbial+Technology%22">Enzyme & Microbial Technology</searchLink>. Jan2016, Vol. 82, p1-7. 7p.
– Name: Subject
  Label: Subjects
  Group: Su
  Data: <searchLink fieldCode="DE" term="%22Operons%22">Operons</searchLink><br /><searchLink fieldCode="DE" term="%22Hydrogenase%22">Hydrogenase</searchLink><br /><searchLink fieldCode="DE" term="%22Genetic+mutation%22">Genetic mutation</searchLink><br /><searchLink fieldCode="DE" term="%22NAD+%28Coenzyme%29%22">NAD (Coenzyme)</searchLink><br /><searchLink fieldCode="DE" term="%22Enterobacter+aerogenes%22">Enterobacter aerogenes</searchLink><br /><searchLink fieldCode="DE" term="%22Hydrogen+production%22">Hydrogen production</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: In this study, the hydrogenase-3 gene cluster ( hycDEFGH ) was isolated and identified from Enterobacter aerogenes CCTCC AB91102. All gene products were highly homologous to the reported bacterial hydrogenase-3 (Hyd-3) proteins. The genes hycE , hycF , hycG encoding the subunits of hydrogenase-3 were targeted for genetic knockout to inhibit the FHL hydrogen production pathway via the Red recombination system, generating three mutant strains AB91102-E (Δ hycE ), AB91102-F (Δ hycF ) and AB91102-G (Δ hycG ). Deletion of the three genes affected the integrity of hydrogenase-3. The hydrogen production experiments with the mutant strains showed that no hydrogen was detected compared with the wild type (0.886 mol/mol glucose), demonstrating that knocking out any of the three genes could inhibit NADH hydrogen production pathway. Meanwhile, the metabolites of the mutant strains were significantly changed in comparison with the wild type, indicating corresponding changes in metabolic flux by mutation. Additionally, the activity of NADH-mediated hydrogenase was found to be nil in the mutant strains. The chemostat experiments showed that the NADH/NAD + ratio of the mutant strains increased nearly 1.4-fold compared with the wild type. The NADH-mediated hydrogenase activity and NADH/NAD + ratio analysis both suggested that NADH pathway required the involvement of the electron transport chain of hydrogenase-3. [ABSTRACT FROM AUTHOR]
– Name: AbstractSuppliedCopyright
  Label:
  Group: Ab
  Data: <i>Copyright of Enzyme & Microbial Technology is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
PLink https://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=egs&AN=111498234
RecordInfo BibRecord:
  BibEntity:
    Identifiers:
      – Type: doi
        Value: 10.1016/j.enzmictec.2015.08.011
    Languages:
      – Code: eng
        Text: English
    PhysicalDescription:
      Pagination:
        PageCount: 7
        StartPage: 1
    Subjects:
      – SubjectFull: Operons
        Type: general
      – SubjectFull: Hydrogenase
        Type: general
      – SubjectFull: Genetic mutation
        Type: general
      – SubjectFull: NAD (Coenzyme)
        Type: general
      – SubjectFull: Enterobacter aerogenes
        Type: general
      – SubjectFull: Hydrogen production
        Type: general
    Titles:
      – TitleFull: Mutational analysis of the hyc-operon determining the relationship between hydrogenase-3 and NADH pathway in Enterobacter aerogenes.
        Type: main
  BibRelationships:
    HasContributorRelationships:
      – PersonEntity:
          Name:
            NameFull: Pi, Jian
      – PersonEntity:
          Name:
            NameFull: Jawed, Muhammad
      – PersonEntity:
          Name:
            NameFull: Wang, Jun
      – PersonEntity:
          Name:
            NameFull: Xu, Li
      – PersonEntity:
          Name:
            NameFull: Yan, Yunjun
    IsPartOfRelationships:
      – BibEntity:
          Dates:
            – D: 01
              M: 01
              Text: Jan2016
              Type: published
              Y: 2016
          Identifiers:
            – Type: issn-print
              Value: 01410229
          Numbering:
            – Type: volume
              Value: 82
          Titles:
            – TitleFull: Enzyme & Microbial Technology
              Type: main
ResultId 1