Kourist Team

H16 mutant PHB (click to enlarge)

Research Goals

Major interest is in hydrolytic enzymes with a focus on esterases, and on lyases. The group has successfully implemented several industrial processes employing hydroxynitrile lyases and esterases.

Besides the molecular characterization, development of methods and strategies for engineering of enzymes by designed evolution is persued. Highly integrated approaches, combining rational and evolutionary routes are followed.

This work includes strong interaction with structural biology and organic chemistry as well as analytical chemistry. Highlights of this research are the identification of an enzyme having hydroxynitrile lyase activity belonging to a novel structural class of proteins and inverting the enantioselectivity of an esterase and of a hydroxynitrile lyase by protein engineering.


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Current Theses

  • Steffen Gruber:
  • Zalina Magomedova:
  • Katharina Mädje: New engineered hydroxynitrile lyases
  • Melanie Hirz:
  • Aline Telzerow: Widening the substrate scope of transaminases by protein engineering
  • Corrina Odar:
  • Michael Lukesch:
  • Felix Tobola:

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Completed Theses

  • Ahmad Mudassar: Development of tools and methods for genetic engineering of the recombinant protein production host Pichia pastoris
  • Thomas Vogl: Synthetic biology to improve protein expression in Pichia pastoris
  • Christine Winkler: Probing the secretory pathway of the yeast Pichia pastoris
  • Anita Emmerstorfer: Improving heterologous cytochrome P450 function in S. cerevisiae
  • Ivan Hajnal: Characterisation, Identification of Novel Functions and Engineering of Substrate Specificity of Cupin Superfamily Enzymes
  • Aleksandra Mitrovic: Discovery and Optimization of Fungal Lignocellulolytic Enzymes
  • Ingund Rosales Rodriguez: Improvement of Pichia pastoris AOX1 promoter expression system
  • Andreas Braun: Expression of human liver cytochrome P450 enzymes in the non-conventional yeast Yarrowia lipolytica for the evaluation of two-liquid biphasic whole-cell biotransformation of steroids
  • Andrea Mellitzer: Expression and engineering of eukaryotic enzymes for polymer modification using Pichia pastoris
  • Martina Geier: Redesign of Oxygenases for Biocatalysis
  • Laura Hannele Näätsaari: Horseradish peroxidase isoenzyme discovery and production in a new improved Pichia pastoris expression system
  • Martin Tengg: Methyltransferases for Novel Biocatalytic C-C Bond Formation
  • Claudia Ruth: Expression of recombinant proteins in Pichia pastoris employing synthetic promoters
  • Brigitte Höller: New Oxidoreductases for Biocatalysis
  • Sandra Abad: New strategies and tools for protein expression and engineering using Pichia pastoris
  • Veeresh Juturu: Enhancing the expression of recombinant proteins in the methylotrophic yeast Pichia pastoris
  • Ulrike Schreiner: Molecular engineering of nitrilases
  • Alexandra Andryushkova: Establishing fission yeast as a host for eukaryotic genomic libraries
  • Maria Elisabeth Freigassner: Alternative oxidases: Elucidation of structural and functional differences between plants and fungi
  • Martin Kietzmann: Pig Liver Esterase: Molecular Development of a Biocatalyst for the Industrial Production of High-Value Pharmaceuticals
  • Beate Pscheidt: Directed Evolution of PaHNL5
  • Matthias Schittmayer: New Oxidoreductases for metabolite synthesis
  • Ruth Breitschädel: Establishment of a new High-Throughput screening system applicable to different hosts and its application for mining endophytic metagenomic libraries
  • Romana Wiedner: Molecular engineering of the industrial biocatalyst hydroxynitrile lyase from Hevea brasiliensis
  • Franz Stefan Hartner: Engineering Pichia pastoris for whole-cell biotransformation
  • Zhibin Liu: Directed Evolution of Industrial Enzymes employing Pichia pastoris
  • Zahid Hussain: Molecular Recruitment of Novel Enzymes from the Endophytic Bacteria
  • Brigitte Pohn: Development of fazile high-throughput screening systems for analyzing enzyme activities
  • Daniel Luschnig: Development of novel bacterial Penicillin V amidases for industrial applications
  • Alexander Kern: Determination of increased yields and productivity in Pichia pastoris by strategic transformation of key genes from Aspergillus niger
  • Roland Weis: High-Throughput Protein Engineering in Pichia Pastoris
  • Sandra Majer: Expression of human growth hormone and human growth hormone binding protein
  • Birgit Brunner: Directed Evolution as a Method to Create Enantioselective Cyclohexanone Monooxygenases

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Selected Publications

2017
Tea Pavkov-Keller, Kerstin Steiner, Mario Faber, Martin Tengg, Helmut Schwab, Mandana Gruber-Khadjawi and Karl Gruber Crystal Structure and Catalytic Mechanism of CouO, a Versatile C-Methyltransferase from Streptomyces rishiriensis Show publication in PURE
2016
Zalina Magomedova, Andreea Grecu, Christoph W Sensen, Helmut Schwab and Petra Heidinger Characterization of two novel alcohol short-chain dehydrogenases/reductases from Ralstonia eutropha H16 capable of stereoselective conversion of bulky substrates Show publication in PURE
Katrin Niegelhell, Michael Süßenbacher, Thomas Ganner, Daniel Schwendenwein, Helmut Schwab, Franz Stelzer, Harald Plank and Stefan. Spirk Enzymes as Biodevelopers for Nano- And Micropatterned Bicomponent Biopolymer Thin Films. Show publication in PURE
Martin Tengg, Harald Stecher, Lisa Offner, Katharina Plasch, Felix Anderl, Hansjörg Weber, Helmut Schwab and Mandana Gruber Methyltransferases Show publication in PURE
2015
Ivan Hajnal, Kurt Faber, Helmut Schwab, Melanie Hall and Kerstin Steiner Oxidative Alkene Cleavage Catalysed by Manganese-​Dependent Cupin TM1459 from Thermotoga maritima Show publication in TUGRAZonline
Thomas Vogl, Mudassar Ahmad, Florian Krainer, Helmut Schwab and Anton Glieder Restriction site free cloning (RSFC) plasmid family for seamless, sequence independent cloning in Pichia pastoris Show publication in TUGRAZonline
Petra Köfinger, Steffen Gruber and Helmut Schwab Versatile plasmid-based expression systems for Gram-negative bacteria-General essentials exemplified with the bacterium Ralstonia eutropha H16. Show publication in TUGRAZonline
Katharina Schmoelzer, Tibor Czabany, Christiane Luley, Tea Pavkov-Keller, Doris Ribitsch, Helmut Schwab, Karl Gruber, Hansjörg Weber and Bernd Nidetzky Complete switch from α-2,3- to α-2,6-regioselectivity in Pasteurella dagmatis β-D-galactoside sialyltransferase by active-site redesign Show publication in TUGRAZonline
Romana Wiedner, Bettina Kothbauer, Tea Pavkov-Keller, Mandana Gruber, Karl Gruber, Helmut Schwab and Kerstin Steiner Improving the Properties of Bacterial R-selective Hydroxynitrile Lyases for Industrial Applications 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
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
Romana Wiedner, Bettina Kothbauer, Tea Pavkov-Keller, Mandana Gruber, Karl Gruber, Helmut Schwab and Kerstin Steiner Improving the Properties of Bacterial R-selective Hydroxynitrile Lyases for Industrial Applications Show publication in PURE
Thomas Vogl, Mudassar Ahmad, Florian Krainer, Helmut Schwab and Anton Glieder Restriction site free cloning (RSFC) plasmid family for seamless, sequence independent cloning in Pichia pastoris Show publication in PURE
Katharina Schmoelzer, Tibor Czabany, Christiane Luley, Tea Pavkov-Keller, Doris Ribitsch, Helmut Schwab, Karl Gruber, Hansjörg Weber and Bernd Nidetzky Complete switch from α-2,3- to α-2,6-regioselectivity in Pasteurella dagmatis β-D-galactoside sialyltransferase by active-site redesign Show publication in PURE
Ivan Hajnal, Kurt Faber, Helmut Schwab, Melanie Hall and Kerstin Steiner Oxidative Alkene Cleavage Catalysed by Manganese-​Dependent Cupin TM1459 from Thermotoga maritima Show publication in PURE
Petra Köfinger, Steffen Gruber and Helmut Schwab Versatile plasmid-based expression systems for Gram-negative bacteria-General essentials exemplified with the bacterium Ralstonia eutropha H16. Show publication in PURE
2014
Steffen Gruber, Jeremias Hagen, Helmut Schwab and Petra Köfinger Versatile and stable vectors for efficient gene expression in Ralstonia eutropha H16 Show publication in TUGRAZonline
Doris Ribitsch, Sabine Zitzenbacher, Peter Andreas Augustin, Katharina Schmoelzer, Christiane Luley, Marco Thomann, Christine Jung, Harald Sobek, Rainer Müller, Bernd Nidetzky and Helmut Schwab High-quality production of human α-2,6-sialyltransferase in Pichia pastoris requires control over N-terminal truncations by host-inherent protease activities Show publication in TUGRAZonline
Katharina Schmoelzer, Christiane Luley, Tibor Czabany, Doris Ribitsch, Helmut Schwab, Hansjörg Weber and Bernd Nidetzky Mechanistic study of CMP-Neu5Ac hydrolysis by α2,3-sialyltransferase from Pasteurella dagmatis Show publication in TUGRAZonline
Romana Wiedner, Mandana Gruber, Helmut Schwab and Kerstin Steiner Discovery of a novel (R)-selective bacterial cupin HNL from Acidobacterium capsulatum ATCC 51196 by a sequence-based approach Show publication in TUGRAZonline
Georg Steinkellner, Christian C. Gruber, Tea Pavkov-Keller, Alexandra Binter, Kerstin Steiner, Christoph Winkler, Andrzej Lyskowski, Orsolya Schwamberger, Monika Oberer, Helmut Schwab, Kurt Faber, Peter Macheroux and Karl Gruber Identification of promiscuous ene-reductase activity by mining structural databases using active site constellations 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
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
Andrzej Franciszek Lyskowski, Christian Gruber, Georg Steinkellner, M Schuermann, Helmut Schwab, Karl Gruber and Kerstin Steiner Crystal structure of an (R)-selective ω-transaminase from Aspergillus terreus (R)-selective ω-transaminase from Aspergillus terre Show publication in TUGRAZonline
Steffen Gruber, Jeremias Hagen, Helmut Schwab and Petra Köfinger Versatile and stable vectors for efficient gene expression in Ralstonia eutropha H16 Show publication in PURE
Katharina Schmoelzer, Christiane Luley, Tibor Czabany, Doris Ribitsch, Helmut Schwab, Hansjörg Weber and Bernd Nidetzky Mechanistic study of CMP-Neu5Ac hydrolysis by α2,3-sialyltransferase from Pasteurella dagmatis 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
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
Georg Steinkellner, Christian C. Gruber, Tea Pavkov-Keller, Alexandra Binter, Kerstin Steiner, Christoph Winkler, Andrzej Lyskowski, Orsolya Schwamberger, Monika Oberer, Helmut Schwab, Kurt Faber, Peter Macheroux and Karl Gruber Identification of promiscuous ene-reductase activity by mining structural databases using active site constellations Show publication in PURE
Doris Ribitsch, Sabine Zitzenbacher, Peter Andreas Augustin, Katharina Schmoelzer, Christiane Luley, Marco Thomann, Christine Jung, Harald Sobek, Rainer Müller, Bernd Nidetzky and Helmut Schwab High-quality production of human α-2,6-sialyltransferase in Pichia pastoris requires control over N-terminal truncations by host-inherent protease activities Show publication in PURE
Romana Wiedner, Mandana Gruber, Helmut Schwab and Kerstin Steiner Discovery of a novel (R)-selective bacterial cupin HNL from Acidobacterium capsulatum ATCC 51196 by a sequence-based approach Show publication in PURE
Andrzej Franciszek Lyskowski, Christian Gruber, Georg Steinkellner, M Schuermann, Helmut Schwab, Karl Gruber and Kerstin Steiner Crystal structure of an (R)-selective ω-transaminase from Aspergillus terreus (R)-selective ω-transaminase from Aspergillus terre Show publication in PURE
2013
Ivan Hajnal, Andrzej Franciszek Lyskowski, Ulf Hanefeld, Karl Gruber, Helmut Schwab and Kerstin Steiner Biochemical and structural characterization of a novel bacterial manganese-dependent hydroxynitrile lyase Show publication in TUGRAZonline
F.G. Eikmeyer, Petra Köfinger, Andrea Poschenel, S Jünemann , M Zakrzewski , Stefan Heinl, Elisabeth Mayrhuber, Reingard Grabherr, Alf Pühler, Helmut Schwab and Andreas Schlüter Metagenome analyses reveal the influence of the inoculant Lactobacillus buchneri CD034 on the microbial community involved in grass ensiling Show publication in TUGRAZonline
Kateryna Fesko, Kerstin Steiner, Rolf Breinbauer, Helmut Schwab, Martin Schürmann and Gernot Strohmeier Investigation of one-enzyme systems in the w-transaminase-catalyzed synthesis of chiral amines Show publication in TUGRAZonline
Katharina Schmoelzer, Doris Ribitsch, Tibor Czabany, Christiane Luley, Deja Kokot, Andrzej Franciszek Lyskowski, Sabine Zitzenbacher, Helmut Schwab and Bernd Nidetzky Characterization of a multifunctional α2,3-sialyltransferase from Pasteurella dagmatis 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
Enrique Herrero Acero, Doris Ribitsch, Anita Dellacher, Sabine Zitzenbacher, Annemarie Marold, Georg Steinkellner, Karl Gruber, Helmut Schwab and Georg Gübitz Surface engineering of a cutinase from Thermobifida cellulosilytica for improved polyester hydrolysis Show publication in TUGRAZonline
Doris Ribitsch, Orcal Yebra Antonio, Sabine Zitzenbacher, Susanne Nowitsch, Georg Steinkellner, Katrin Julia Greimel, Doliška Aleš, Gustav Oberdorfer, Christian Gruber, Karl Gruber, Helmut Schwab, Stana-Kleinschek Karin, Enrique Herrero Acero and Georg Gübitz Fusion of binding-domains to Thermobifida cellulosilytica cutinase to tune sorption characteristics and enhancing PET hydrolysis Show publication in TUGRAZonline
2012
Stefan Heinl, D Wibberg, F.G. Eikmeyer, R Szczepanowski , J Blom, B Linke, A Goesmann, Reingard Grabherr, Helmut Schwab, Alf Pühler and Andreas Schlüter Insights into the completely annotated genome of Lactobacillus buchneri CD034, a strain isolated from stable grass silage Show publication in TUGRAZonline
Kerstin Steiner and Helmut Schwab Recent advances in rational approaches for enzyme engineerings in rational approaches for enzyme engineering Show publication in TUGRAZonline
Enrique Herrero Acero, Doris Ribitsch, Katrin Julia Greimel, Anita Dellacher, Sabine Zitzenbacher, Annemarie Marold, Rosario Diaz Rodriguez, Gibson Stephen Nyanhongo, Helmut Schwab and Georg Gübitz Two-step enzymatic functionalisation of polyamide with phenolics Show publication in TUGRAZonline
Andrzej Lyskowski, Martin Tengg, Georg Steinkellner, Helmut Schwab, Mandana Gruber and Karl Gruber Crystallisation of the novel S-adenosyl-L-methionine dependent C-methyltransferase CouO from Streptomyces rishiriensis and preliminary diffraction data analysis. Acta Crystallogr., Sect. F: Struct. Biol. Cryst. Commun. Show publication in TUGRAZonline
Martin Tengg, Harald Stecher, Peter Remler, Inge Eiteljörg, Mandana Gruber and Helmut Schwab Molecular characterization of the C-Methyltransferase NovO of Streptomyces spheroides, a valuable enzyme for performing Friedel-Crafts alkylation Show publication in TUGRAZonline
Doris Ribitsch, Enrique Herrero Acero, Katrin Julia Greimel, Anita Dellacher, Sabine Zitzenbacher, Annemarie Marold, Rosario Diaz Rodriguez, Georg Steinkellner, Karl Gruber, Helmut Schwab and Georg Gübitz A New Esterase from Thermobifida halotolerans Hydrolyses Polyethylene Terephthalate (PET) and Polylactic Acid (PLA) Show publication in TUGRAZonline
Kerstin Steiner, Andrzej Franciszek Lyskowski, Ivan Hajnal, Georg Steinkellner, Helmut Schwab and Karl Gruber Crystallization of a novel metal-containing cupin from Acidobacterium sp. and preliminary diffraction data analysis Show publication in TUGRAZonline
Doris Ribitsch, Enrique Herrero Acero, Katrin Julia Greimel, Inge Eiteljörg, Eva Trotscha, Giuliano Freddi, Helmut Schwab and Georg Gübitz Characterization of a new cutinase from Thermobifida alba regarding PET-surface hydrolysis Show publication in TUGRAZonline
Zahid Hussain, Romana Wiedner, Kerstin Steiner, Tanja Hajek, Manuela Avi , Bianca Hecher, Angela Sessitsch and Helmut Schwab Characterization of two bacterial hydroxynitrile lyases with high similarity to cupin superfamily proteins Show publication in TUGRAZonline
Doris Ribitsch, Sonja Heumann, W. Karl, Jochen Gerlach, R. Leber, Ruth Birner-Grünberger, Karl Gruber, Inge Eiteljörg, Peter Remler, P. Siegert, J. Lange, Karl-Heinz Maurer, Gabriele Berg, Georg Gübitz and Helmut Schwab Extracellular serine proteases from Stenotrophomonas maltophilia: screening, isolation and heterologous expression in E.coli. Show publication in TUGRAZonline
2011
Enrique Herrero Acero, Doris Ribitsch, Georg Steinkellner, Karl Gruber , Katrin Julia Greimel, Inge Eiteljörg, Eva Trotscha, R. Wei, W. Zimmermann, M. Zinn, A. Cavaco-Paulo, G. Freddi, Helmut Schwab and Georg Gübitz Enzymatic Surface Hydrolysis of PET. Effect of Structural Diversity on Kinetic Properties of Cutinases from Thermobifida Show publication in TUGRAZonline
Doris Ribitsch, Sonja Heumann, Eva Trotscha, Enrique Herrero Acero, Katrin Julia Greimel, Inge Eiteljörg, Peter Remler, Regina Maria Leber, Sigrid Deller, Giuliano Freddi, I. Donelli, K.H. Maurer, P. Siegert, T. Weber, Helmut Schwab and Georg Gübitz Hydrolysis of polyethyleneterephthalate by para-nitrobenzylesterase from Bacillus subtilis 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
2010
Doris Ribitsch, W Karl, Ruth Birner-Grünberger, Karl Gruber, Inge Eiteljörg, Peter Remler, Susanne Wieland, Petra Siegert, Karl Maurer and Helmut Schwab Engineering of choline oxidase from Arthrobacter nicotianae for potential use as biological bleach in detergents. Show publication in TUGRAZonline
Doris Ribitsch, W Karl, Ruth Birner-Grünberger, Karl Gruber, Inge Eiteljörg, Peter Remler, Susanne Wieland, Petra Siegert, Karl Maurer and Helmut Schwab C-terminal truncation of a metagenome-derived detergent protease for effective expression in E. coli. Show publication in TUGRAZonline

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Research group leader
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Robert Kourist
Univ.-Prof. Dr.rer.nat.
Office hours: Wednesday, 9:00-11:00 (room BKEG05058)