Pichler Team

October 2013
October 2013
March 2015
March 2015

Research Goals

We are interested in processes that involve biological membranes. We engineer the membranes of microbes, i.e. bacteria and yeasts, regarding lipid and membrane protein composition, either to understand the complex interactions of these hydrophobic compounds or to apply the modified microorganisms for interesting goals of industrial biotechnology. Our particular focus in the yeast lipid field are sterols, their biosynthesis and homeostasis, and their interactions with membrane proteins.

Pichia pastoris, a methylotrophic yeast, is our favourite host, which we employ for expression and secretion of recombinant soluble proteins as well as for the biosynthesis of valuable metabolites, e.g. plant terpenoids, through heterologous expression of membrane proteins. We have uncovered several endogenous yeast proteins and mechanisms that render protein secretion or membrane protein expression more efficient and sustainable. Based on our expertise in medium to high-throughput screening approaches involving microbes and a background in yeast cell biology, we expand the knowledge on membrane-related processes and extend the molecular toolboxes for applications on industrial scale.

Rational and evolutionary enzyme engineering approaches allow us to make novel enzymes or enzyme families fit for biocatalytic applications, as isolated enzymes or in whole-cell applications. Therefore, we study enzyme characteristics by classical protein analytics and devise novel enzyme assays. We usually team up with structural biologists, biochemists as well as analytical and organic chemists.


To top

Current Projects

  • January 2014 – ongoing
    ACIB projects 4.5-6 and 24.011 - Engineering hydratases

    We are engineering hydratases for industrial fitness. This project is carried out with company partners at DSM Chemical Technology R&D B.V., Innovative Synthesis (Geleen, The Netherlands).


  • February 2015 – ongoing
    Industrial collaboration project - Terpenoid screening

    We are developing a medium for a high throughput terpenoid screening procedure in collaboration with Isobionics (Geleen, The Netherlands).


To top

Past Projects

  • September 2005 – June 2009
    Applied Biocatalysis projects 2.11 and P09 - Recombinant esterase for alpha-substituted aliphatic esters – improving the yield

    We have identified a novel isoform of the pig liver esterase protein family, markedly improved expression of pig liver esterases in P. pastoris and E. coli, and have evolutionarily engineered pig liver esterases for biocatalytic applications. This project has been carried out in close collaboration with Prof. Helmut Schwab, Prof. Karl Gruber and company partners at DSM Chemical Technology R&D B.V., Innovative Synthesis (Geleen, The Netherlands) and DPx Fine Chemicals (Linz, Austria).


  • July 2009 – December 2013
    ACIB projects PÜ4.1 and 4.2-1 - Enzyme engineering for complex reactions

    We have generated a promising yeast whole-cell biocatalyst that makes (+)-nootkatone from simple carbon sources. To foster plant terpenoid biosynthesis in yeasts, and to improve the industrial applicability of membrane-attached cytochrome P450 enzymes we have identified several endogenous helper factors in P. pastoris and S. cerevisiae. This project has been carried out in close collaboration with company partners at DSM Chemical Technology R&D B.V., Innovative Synthesis (Geleen, The Netherlands) and DPx Fine Chemicals (Linz, Austria).


  • October 2009 – April 2014
    DK Molecular Enzymology (FWF) PhD project Dr. Christine Winkler - Probing the secretory pathway of the yeast Pichia pastoris

    We have devised a novel, medium-throughput, random mutagenesis/screening protocol for improved protein secretion in P. pastoris. A couple of highly interesting target genes have been identified, partially characterized and filed for patenting. This project has been carried out in close collaboration with Prof. Helmut Schwab.


  • January 2010 – December 2015
    ACIB project 3.2-1 - Pichia pastoris cell factory for protein production

    We have applied multiple approaches to identify key genes that determine protein secretion rates in P. pastoris. Upregulation of several target genes has improved recombinant protein titers and yields. This project has been carried out in close collaboration with Prof. Diethard Mattanovich (Vienna), Prof. Guenther Daum and company partners at Biocrates (Innsbruck, Austria), Biomin (Tulln, Austria), Boehringer Ingelheim-RCV (Vienna, Austria), LONZA (Visp, Switzerland), Sandoz (Kundl, Austria) and VTU (Grambach, Austria).


  • January 2010 – June 2015
    ACIB projects 4.2-2 and 54.041 - Enzyme engineering and substrate channeling for vitamin production

    We have developed and employed screenings for improved production of vitamin precursors. Remarkably, we have managed to engineer a very complex enzyme that has evaded structural elucidation to date. This project was carried out in close collaboration with company partners at DSM Nutritional Products (Kaiseraugst, Switzerland).


  • January 2015 – February 2017
    ACIB project 23.021 - Engineering of protein secretion in Pichia pastoris

    We were following up on several approaches to enhance recombinant protein titers and yields. This project was carried out in close collaboration with Prof. Diethard Mattanovich (Vienna), Prof. Guenther Daum and company partners at Biomin (Tulln, Austria), Boehringer Ingelheim-RCV (Vienna, Austria), LONZA (Visp, Switzerland), Sandoz (Kundl, Austria) and VTU (Grambach, Austria).


To top

Current Theses

  • PhD Theses

    • Matthias Engleder
      Engineering hydratases
    • Melanie Hirz
      Cell engineering in Pichia pastoris
    • Sandra Moser
      Triterpenoid synthesis in yeast
  • Master Theses

    • Carolin Kunz
    • Alena Voit
    • Philippe Oberhemm
    • Edmund Dittmann
    • Kerstin Stadler
    • Klara Lanz
    • Martin Senekowitsch
    • Katharina Ebner
    • Johannes Blamauer
    • Zan Caf-Feldin
    • Chiam Hashem
    • Katharina Nagl
    • Felix Stocker-Reicher
    • Harald Blasl
    • Johanna Zwick
    • Sanja Tunjic

To top

Completed Theses

  • PhD Theses

    • Dr. Martin Trinker (né Kietzmann)
      January 2009
      Pig liver esterase: Molecular development of a biocatalyst for the industrial production of high-value pharmaceuticals
    • Dr. Veeresh Juturu
      June 2010
      Enhancing the expression of recombinant proteins in the methylotrophic yeast Pichia pastoris
    • Dr. Anita Emmerstorfer
      April 2014
      Improving heterologous cytochrome P450 function in S. cerevisiae
    • Dr. Christine Winkler
      April 2014
      Probing the secretory pathway of the yeast Pichia pastoris
  • Master Theses

    • 2005
      Dr. Veeresh Juturu
    • 2007
      Anja Egger Dr. Michael Tscherner Dr. Eva Hoffmann
    • 2008
      Dr. Andreas Braun Dr. Julia Feichtinger
    • 2009
      Dr. Amin El-Heliebi Dr. Christine Winkler 
    • 2010
      Dr. Andreas Küberl Michael Felber Dr. Anita Emmerstorfer Dr. Birgit Ploier
    • 2011
      Dr. Thomas Vogl Dr. Corinna Odar Peter Augustin Gerald Richter
    • 2012
      Melanie Hirz
    • 2013
      Ariane Fankl Peter Neugebauer
    • 2014
      Katharina Littringer Sandra Moser Miriam Wimmer-Teubenbacher
    • 2015
      Matthias Engleder Florian Ungerböck Johannes Fuchs
      Sabine Schrempf Bernd Albrecht Mia Potocnjak
    • 2016
      Simon Arhar Stefanie Gabriel Manfred Briendl Elisabeth Hirschböck
    • 2017
      Birgit Grill Christoph Schilling Melissa Horvat Valentin Steinwender

To top

Partners

Scientific partners

  • Prof. Erich Leitner - Institute of Analytical and Food Chemistry, Graz University of Technology, Austria
  • Dr. Gernot Strohmeier - Institute of Organic Chemistry, Graz University of Technology, Austria
  • Prof. Guenther Zellnig - Institute of Plant Physiology, University of Graz, Austria
  • Prof. Harro Bouwmeester - Laboratory of Plant Physiology, University of Wageningen, The Netherlands
  • Prof. Howard Riezman - Department of Biochemistry, University of Geneva, Switzerland
  • Prof. Karl Gruber - Institute of Molecular Biosciences, University of Graz, Austria
  • Prof. Koichi Funato - Department of Biofunctional Science and Technology, Graduate School of Biosphere Science Hiroshima University, Japan
  • Prof. Maria A Vanoni - Department of Biomolecular Sciences and Biotechnology, University of Milano, Italy
  • Dr. Michael Haertlein - Institut Laue-Langevin, Grenoble, France
  • Prof. Steven JD Karlish - Department of Biological Chemistry, Weizmann Institute of Science, Rehovot, Israel

Industrial collaborations

  • ACIB, Austrian Centre of Industrial Biotechnology (Graz, Vienna, Innsbruck, Austria)
  • DSM Chemical Technology R&D B.V., Innovative Synthesis (Geleen, The Netherlands)
  • DPx Fine Chemicals (Linz, Austria)
  • Biomin (Tulln, Austria)
  • Boehringer Ingelheim RCV (Vienna, Austria) 
  • LONZA (Visp, Switzerland)
  • Sandoz (Kundl, Austria)
  • VTU (Grambach, Austria)

To top

Selected Publications

2016
Anita Emmerstorfer and Harald Pichler Production of aromatic plant terpenoids in recombinant baker´s yeastBiotechnology of Plant Secondary Metabolism79 - 89 Show publication in TUGRAZonline
Harald Pichler Oleate hydratase - structure function relationships and applicationOleate hydratase - structure function relationships and application33 Show publication in PURE
Anita Emmerstorfer, Sandra Moser and Harald Pichler Screening for improved isoprenoid biosynthesis in microorganisms Show publication in PURE
Tamara Wriessnegger, Sandra Moser, Anita Emmerstorfer-Augustin, Erich Leitner, Monika Müller, Iwona Kaluzna, Martin Schürmann, Mink Daniel and Harald Pichler Enhancing cytochrome P450-mediated conversions in P. pastoris through RAD52 over-expression and optimizing the cultivation conditions Show publication in PURE
Anita Emmerstorfer and Harald Pichler Production of aromatic plant terpenoids in recombinant baker´s yeastProduction of aromatic plant terpenoids in recombinant baker´s yeast79-89 Show publication in PURE
2015
Matthias Engleder, Tea Pavkov-Keller, Anita Emmerstorfer, Hromic Altijana, Sabine Schrempf, Georg Steinkellner, Tamara Wriessnegger, Erich Leitner, Gernot Strohmeier, Iwona Kaluzna, Mink Daniel, Schürmann Martin, Silvia Wallner, Peter Macheroux, Karl Gruber and Harald Pichler Structure-based mechanism of oleate hydratase from Elizabethkingia meningoseptica Show publication in TUGRAZonline
Atsuko Ikeda, Tetsuya Muneoka, Suguru Murakami, Ayaka Hirota, Yukari Yabuki , Takefumi Karashima, Kota Nakazono, Masahiro Tsuruno, Harald Pichler, Katsuhiko Shirahige, Yukiko Kodama, Toshi Shimamoto, Keiko Mizuta and Kouichi Funato Sphingolipids regulate telomere clustering by affecting transcriptional levels of genes involved in telomere homeostasis Show publication in TUGRAZonline
Anita Emmerstorfer, Miriam Wimmer-Teubenbacher, Tamara Wriessnegger, Erich Leitner, Monika Müller, Iwona Kaluzna, Martin Schürmann, Daniel Mink, Günther Zellnig, Helmut Schwab and Harald Pichler Over-expression of ICE2 stabilizes cytochrome P450 reductase levels in Saccharomyces cerevisiae and Pichia pastoris Show publication in TUGRAZonline
Matthias Engleder, Tea Pavkov-Keller, Anita Emmerstorfer, Hromic Altijana, Sabine Schrempf, Georg Steinkellner, Tamara Wriessnegger, Erich Leitner, Gernot Strohmeier, Iwona Kaluzna, Mink Daniel, Schürmann Martin, Silvia Wallner, Peter Macheroux, Karl Gruber and Harald Pichler Structure-based mechanism of oleate hydratase from Elizabethkingia meningoseptica Show publication in PURE
Anita Emmerstorfer, Miriam Wimmer-Teubenbacher, Tamara Wriessnegger, Erich Leitner, Monika Müller, Iwona Kaluzna, Martin Schürmann, Daniel Mink, Günther Zellnig, Helmut Schwab and Harald Pichler Over-expression of ICE2 stabilizes cytochrome P450 reductase levels in Saccharomyces cerevisiae and Pichia pastoris Show publication in PURE
Atsuko Ikeda, Tetsuya Muneoka, Suguru Murakami, Ayaka Hirota, Yukari Yabuki, Takefumi Karashima, Kota Nakazono, Masahiro Tsuruno, Harald Pichler, Katsuhiko Shirahige, Yukiko Kodama, Toshi Shimamoto, Keiko Mizuta and Kouichi Funato Sphingolipids regulate telomere clustering by affecting transcriptional levels of genes involved in telomere homeostasis Show publication in PURE
2014
Claudia Ruth, Markus Buchetics , Viktorija Vidimce, Daniela Kotz, Stefan Naschberger, Diethard Mattanovich, Harald Pichler and Brigitte Gasser Pichia pastoris Aft1 - a novel transcription factor, enhancing recombinant protein secretion Show publication in TUGRAZonline
Anita Emmerstorfer, Tamara Wriessnegger, Melanie Hirz and Harald Pichler Overexpression of membrane proteins from higher eukaryotes in yeasts Show publication in TUGRAZonline
Tamara Wriessnegger, Peter Andreas Augustin, Matthias Engleder, Erich Leitner, Monika Müller, Iwona Kaluzna, Martin Schürmann, Daniel Mink, Günther Zellnig, Helmut Schwab and Harald Pichler Production of the sesquiterpenoid (+)-nootkatone by metabolic engineering of Pichia pastors Show publication in TUGRAZonline
Michael Felber, Harald Pichler and Claudia Ruth Strains and Molecular Tools for Recombinant Protein Production in Pichia pastorisYeast Metabolic Engineering: Methods and Protocols87 - 111 Show publication in TUGRAZonline
Ahmad Mudassar, Melanie Hirz, Harald Pichler and Helmut Schwab Protein expression in Pichia pastoris: recent achievements and perspectives for heterologous protein production Show publication in TUGRAZonline
Anita Emmerstorfer, Tamara Wriessnegger, Melanie Hirz and Harald Pichler Overexpression of membrane proteins from higher eukaryotes in yeasts Show publication in PURE
Tamara Wriessnegger, Peter Andreas Augustin, Matthias Engleder, Erich Leitner, Monika Müller, Iwona Kaluzna, Martin Schürmann, Daniel Mink, Günther Zellnig, Helmut Schwab and Harald Pichler Production of the sesquiterpenoid (+)-nootkatone by metabolic engineering of Pichia pastors Show publication in PURE
Claudia Ruth, Markus Buchetics, Viktorija Vidimce, Daniela Kotz, Stefan Naschberger, Diethard Mattanovich, Harald Pichler and Brigitte Gasser Pichia pastoris Aft1 - a novel transcription factor, enhancing recombinant protein secretion Show publication in PURE
Ahmad Mudassar, Melanie Hirz, Harald Pichler and Helmut Schwab Protein expression in Pichia pastoris: recent achievements and perspectives for heterologous protein production Show publication in PURE
Michael Felber, Harald Pichler and Claudia Ruth Strains and Molecular Tools for Recombinant Protein Production in Pichia pastorisStrains and Molecular Tools for Recombinant Protein Production in Pichia pastoris87-111 Show publication in PURE
2013
Aashrita Rajagopalan, Markus Schober, Anita Emmerstorfer, Lucas Hammerer, Anna Migglautsch, Birgit Seisser, Silvia M Glück, Frank Niehaus, Jürgen Eck, Harald Pichler, Karl Gruber and Wolfgang Kroutil Enzymatic aerobic alkene cleavage catalyzed by a Mn3+-dependent proteinase A homologue. Show publication in TUGRAZonline
Christine Winkler, Mudassar Ahmad, Peter Neugebauer, Helmut Schwab and Harald Pichler Probing the secretory pathway of the yeast Pichia pastorisBook of abstracts38 - 38 Show publication in TUGRAZonline
Tamara Wriessnegger, Anita Emmerstorfer, Monika Müller, Iwona Kaluzna, Helmut Schwab and Harald Pichler Terpenoid production in yeastsBook of abstracts26 - 26 Show publication in TUGRAZonline
Melanie Hirz, Gerald Richter, Erich Leitner, Tamara Wriessnegger and Harald Pichler A novel cholesterol-producing Pichia pastoris strain is an ideal host for functional expression of human Na,K-ATPase α3β1 isoform Show publication in TUGRAZonline
Tamara Wriessnegger and Harald Pichler Yeast metabolic engineering - targeting sterol metabolism and terpenoid formation Show publication in TUGRAZonline
Aashrita Rajagopalan, Markus Schober, Anita Emmerstorfer, Lucas Hammerer, Anna Migglautsch, Birgit Seisser, Silvia M Glück, Frank Niehaus, Jürgen Eck, Harald Pichler, Karl Gruber and Wolfgang Kroutil Enzymatic aerobic alkene cleavage catalyzed by a Mn3+-dependent proteinase A homologue. Show publication in PURE
Tamara Wriessnegger and Harald Pichler Yeast metabolic engineering - targeting sterol metabolism and terpenoid formation Show publication in PURE
Melanie Hirz, Gerald Richter, Erich Leitner, Tamara Wriessnegger and Harald Pichler A novel cholesterol-producing Pichia pastoris strain is an ideal host for functional expression of human Na,K-ATPase α3β1 isoform Show publication in PURE
Tamara Wriessnegger, Anita Emmerstorfer, Monika Müller, Iwona Kaluzna, Helmut Schwab and Harald Pichler Terpenoid production in yeastsTerpenoid production in yeasts26-26 Show publication in PURE
Christine Winkler, Mudassar Ahmad, Peter Neugebauer, Helmut Schwab and Harald Pichler Probing the secretory pathway of the yeast Pichia pastorisProbing the secretory pathway of the yeast Pichia pastoris38-38 Show publication in PURE
2012
Martina Geier, Andreas Braun, Anita Emmerstorfer, Harald Pichler and Anton Glieder Production of human cytochrome P450 2D6 drug metabolites with recombinant microbes--a comparative study Show publication in TUGRAZonline
Harald Pichler and Tamara Wriessnegger Pichia pastoris as whole-cell biocatalyst for (+)-nootkatone productionPichia 201248 - 48 Show publication in TUGRAZonline
Martina Geier, Andreas Braun, Anita Emmerstorfer, Harald Pichler and Anton Glieder Production of human cytochrome P450 2D6 drug metabolites with recombinant microbes--a comparative study Show publication in PURE
Harald Pichler and Tamara Wriessnegger Pichia pastoris as whole-cell biocatalyst for (+)-nootkatone productionPichia pastoris as whole-cell biocatalyst for (+)-nootkatone production48-48 Show publication in PURE
2011
Cleiton M Souza, Tatjana M Schwabe, Harald Pichler, Birgit Ploier, Erich Leitner, Xue Li Guan, Markus W Wenk, Isabelle Riezman and Howard Riezman A stable yeast strain efficiently producing cholesterol instead of ergosterol is functional for tryptophan uptake, but not weak organic acid resistance Show publication in TUGRAZonline
Gernot Strohmeier, Harald Pichler, Oliver May and Mandana Gruber Application of Designed Enzymes in Organic Synthesis Show publication in TUGRAZonline
Andreas Küberl, Jessica Schneider, Gerhard Thallinger, Ingund Anderl, Daniel Wibberg, Tanja Hajek, Sebastian Jaenicke, Karina Brinkrolf, Alexander Goesmann, Rafael Szczepanowski, Alfred Pühler, Helmut Schwab, Anton Glieder and Harald Pichler High-quality genome sequence of Pichia pastoris CBS7435 Show publication in TUGRAZonline
Gernot Strohmeier, Harald Pichler, Oliver May and Mandana Gruber Application of Designed Enzymes in Organic Synthesis Show publication in PURE
Cleiton M Souza, Tatjana M Schwabe, Harald Pichler, Birgit Ploier, Erich Leitner, Xue Li Guan, Markus W Wenk, Isabelle Riezman and Howard Riezman A stable yeast strain efficiently producing cholesterol instead of ergosterol is functional for tryptophan uptake, but not weak organic acid resistance Show publication in PURE
Andreas Küberl, Jessica Schneider, Gerhard Thallinger, Ingund Anderl, Daniel Wibberg, Tanja Hajek, Sebastian Jaenicke, Karina Brinkrolf, Alexander Goesmann, Rafael Szczepanowski, Alfred Pühler, Helmut Schwab, Anton Glieder and Harald Pichler High-quality genome sequence of Pichia pastoris CBS7435 Show publication in PURE

To top

Team Leader
image/svg+xml

Harald Pichler
Assoc.Prof. Dipl.-Ing. Dr.techn.