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@ARTICLE{Rahmen:280732,
      author       = {Rahmen, N. and Schlupp, C. D. and Mitsunaga, H. and Fulton,
                      Alexander and Aryani, T. and Esch, L. and Schaffrath, U. and
                      Fukuzaki, E. and Jaeger, Karl-Erich and Büchs, J.},
      title        = {{A} particular silent codon exchange in a recombinant gene
                      greatly influences host cell metabolic activity},
      journal      = {Microbial cell factories},
      volume       = {14},
      issn         = {1475-2859},
      address      = {London},
      publisher    = {Biomed Central},
      reportid     = {FZJ-2016-00489},
      pages        = {156},
      year         = {2015},
      abstract     = {BackgroundRecombinant protein production using Escherichia
                      coli as expression host is highly efficient, however, it
                      also induces strong host cell metabolic burden. Energy and
                      biomass precursors are withdrawn from the host’s
                      metabolism as they are required for plasmid replication,
                      heterologous gene expression and protein production. Rare
                      codons in a heterologous gene may be a further drawback.
                      This study aims to investigate the influence of particular
                      silent codon exchanges within a heterologous gene on host
                      cell metabolic activity. Silent mutations were introduced
                      into the coding sequence of a model protein to introduce all
                      synonymous arginine or leucine codons at two randomly
                      defined positions, as well as substitutions leading to
                      identical amino acid exchanges with different synonymous
                      codons. The respective E. coli clones were compared during
                      cultivation in a mineral autoinduction medium using
                      specialized online and offline measuring techniques to
                      quantitatively analyze effects on respiration, biomass and
                      protein production, as well as on carbon source consumption,
                      plasmid copy number, intracellular nucleobases and mRNA
                      content of each clone.ResultsHost stain metabolic burden
                      correlates with recombinant protein production. Upon
                      heterologous gene expression, tremendous differences in
                      respiration, biomass and protein production were observed.
                      According to their different respiration activity the E.
                      coli clones could be classified into two groups, Type A and
                      Type B. Type A clones tended to higher product formation,
                      Type B clones showed stronger biomass formation. Whereas
                      codon usage and intracellular nucleobases had no influence
                      on the Type-A–Type-B-behavior, plasmid copy number, mRNA
                      content and carbon source consumption strongly differed
                      between the two groups.ConclusionsParticular silent codon
                      exchanges in a heterologous gene sequence led to differences
                      in initial growth of Type A and Type B clones. Thus, the
                      biomass concentration at the time point of induction varied.
                      In consequence, not only plasmid copy number and expression
                      levels differed between the two groups, but also the
                      kinetics of lactose and glycerol consumption. Even though
                      the underlying molecular mechanisms are not yet identified
                      we observed the astonishing phenomenon that particular
                      silent codon exchanges within a heterologous gene
                      tremendously affect host cell metabolism and recombinant
                      protein production. This could have great impact on codon
                      optimization of heterologous genes, screening procedures for
                      improved variants, and biotechnological protein production
                      processes.},
      cin          = {IMET},
      ddc          = {610},
      cid          = {I:(DE-Juel1)IMET-20090612},
      pnm          = {89581 - Biotechnology (POF2-89581)},
      pid          = {G:(DE-HGF)POF2-89581},
      typ          = {PUB:(DE-HGF)16},
      UT           = {WOS:000362283400004},
      doi          = {10.1186/s12934-015-0348-8},
      url          = {https://juser.fz-juelich.de/record/280732},
}