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@PHDTHESIS{Zambanini:686218,
      author       = {Zambanini, Thiemo},
      othercontributors = {Blank, Lars M. and Schirawski, Jan},
      title        = {{W}hat can we do with smut? {O}rganic acid production from
                      glycerol with {U}stilaginaceae; 1. {A}uflage},
      volume       = {5},
      school       = {RWTH Aachen University},
      type         = {Dissertation},
      address      = {Aachen},
      publisher    = {Apprimus Verlag},
      reportid     = {RWTH-2017-02718},
      isbn         = {978-3-86359-492-3},
      series       = {Applied microbiology},
      pages        = {XV, 132 Seiten : Illustrationen, Diagramme},
      year         = {2017},
      note         = {Druckausgabe: 2017. - Auch veröffentlicht auf dem
                      Publikationsserver der RWTH Aachen University; Dissertation,
                      RWTH Aachen University, 2016},
      abstract     = {The rapidly growing need for energy and fuel combined with
                      an increased awareness of the detrimental influence of
                      fossil resources has evoked the investigation of renewable
                      alternatives, such as biodiesel. The production process of
                      biodiesel, however, results in a huge waste stream of crude
                      glycerol, reducing the economic and ecological advantage.The
                      overall aim of this thesis was to establish a production
                      process for organic acids from biodiesel derived glycerol
                      with Ustilaginaceae. The biodiversity within this family is
                      well known and in a screening of 126 Ustilaginaceae, two
                      promising candidates for organic acid production were found
                      - Ustilago trichophora for malic acid production and U.
                      vetiveriae for itaconic acid production. Glycerol uptake and
                      growth rate of both strains were improved by adaptive
                      laboratory evolution. Selection of the best growing single
                      colony for each strain and medium and process optimization
                      drastically improved the production values.The itaconic acid
                      titer of U. vetiveriae TZ1 was increased to 35 g L 1
                      produced at a production rate of 0.09 g L 1 h 1.
                      Simultaneously about 60 g L 1 malic acid were formed. In
                      first metabolic engineering approaches overexpressing the
                      mitochondrial transporter Mtt1 and the regulator of the
                      itaconic acid gene cluster Ria1, both from U. maydis, the
                      production values could be shifted in favor of itaconic acid
                      increasing it by 1.5 and 2.0 fold, respectively.
                      Simultaneously, the malic acid titer was reduced by 96 $\%$
                      and 61 $\%,$ respectively.For U. trichophora TZ1 the malic
                      acid titer was improved to nearly 200 g L 1 produced within
                      264 h reaching a maximal production rate of 1.53 g L 1 h 1.
                      Since the knowledge on this obscure U. trichophora was
                      scarce, potential native target genes for metabolic
                      engineering were identified after de novo genome sequencing.
                      To enable the improvement of malic acid production with U.
                      trichophora TZ1 by metabolic engineering, existing tools,
                      such as antibiotic markers and promoters, were investigated
                      and adapted to be suitable for creation of overexpression
                      mutants. Using these tools, overexpression mutants for two
                      different malate dehydrogenases (Mdh1 and Mdh2),
                      pyruvate-carboxylase (Pyc) and two different malic acid
                      transporters (Ssu1 and Ssu2) were generated. While
                      overexpression of Pyc did not improve malic acid production,
                      transformants overexpressing Mdh1 and Mdh2 and malic acid
                      transporters Ssu1 and Ssu2 showed an up to 38 $\%$ increased
                      malic acid production rate and an up to 54 $\%$ increased
                      yield in shake flasks compared to U. trichophora TZ1. In
                      bioreactor cultivations with the mutant overexpressing Ssu2,
                      an increased production rate could not be observed. Due to a
                      drastically lowered optical density, however, this strain
                      had a 29 $\%$ higher specific production rate. Additionally,
                      the product yield was improved by 1.4 fold.These results
                      clearly strengthen the applicability of Ustilaginaceae as
                      industrially valuable production organisms. By this
                      valorization of biodiesel derived crude glycerol, the
                      overall biodiesel bio refinery concept is improved on an
                      economic as well as ecological level.},
      cin          = {161710 / 160000},
      ddc          = {570},
      cid          = {$I:(DE-82)161710_20140620$ / $I:(DE-82)160000_20140620$},
      pnm          = {031A217F - Strategische Allianz ZeroCarbFP - Funktionale
                      Biomasse aus kohlenstoffreichen Abfallströmen, TP 6},
      pid          = {G:(DE-82)BMBF-031A217F},
      typ          = {PUB:(DE-HGF)3 / PUB:(DE-HGF)11},
      doi          = {10.18154/RWTH-2017-02718},
      url          = {https://publications.rwth-aachen.de/record/686218},
}