Winkler Team

Group photo July 2017

Research Goals

Margit Winkler is fascinated by the power of natural biocatalysts for chemical synthesis. Many of the reactions that one may want to use for the preparation of organic molecules have not yet been developed, or have some limitations such as: the need for highly complicated and expensive catalysts, multiple steps or they simply suffer from low selectivity.

The overall theme of Margit's research is to find new and alternative ways to catalyse a transformation by using enzymes. Sometimes, the goal is to determine the scope and limitations of an enzyme and possibly its engineering towards desired characteristics to overcome these limitations. Other times, it is the discovery and detailed study of enzymes that are not well known yet, in order to understand them better and to determine whether and how they could become applicable for synthesis. And finally, a particular product may be in the focus of the studies and the goal is then to find the best way to prepare it.

Due to this broad portfolio of interests and needs, enzymes, substrates, products and methods that have been in the focus of the team over the years, it is essential to collaborate vividly and connect the knowledge of diverse fields. 


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Current Projects

  • May 2017 - April 2020 FFG Industrienahe Dissertation:  Carboxylic Acid Reductases for Enzymatic Reduction of organic Acids to Aldehydes (CARERAA)

    In this project, we investigate CARs in depth with a focus on process relevant parameters.


  • January 2107 - December 2018 - OeAD Project ZA 17/2017: Plant hydroxynitrile lyase diversity and application in industrial biocatalysis

    The goal of this project is to discover new enzymes from South African resources in collaboration with Prof. Karl Rumbold, University of the Witwatersrand, Johannesburg, South Africa.


  • November 2015 - October 2018 - FWF Project P 28477-B21: Catalytic cascades for selective aldehyde syntheses in vivo

    The main goal of this project is to understand the requirements and limitations of microbial cells for aldehyde synthesis. The synthesis of target products– chiral fragrance aldehydes – will be tackled by a combination of chemo-catalysis with a multi-enzymatic in vivo cascade reaction. This is a joint project with Dr. Florian Rudroff, Institute of Applied Synthetic Chemistry, TUVienna

    Project website: http://www.invivocascades.com


  • March 2015 - February 2019 - FWF Elise-Richter-Fellowship V415-B21: Biocatalytic Carboxylate reduction


     

    The aim of this project is to get a better understanding of carboxylate reducing enzymes to enable the development of cell factories for the selective enzymatic preparation of aldehydes. We would like to know more about the structure-function relationship of this enzyme class and to identify more of these enzymes on amino acid level.


  • April 2015 - March 2019 - Horizon2020 Project ROBOX: Grant agreement #635734



    The project title is Expanding the industrial use of Robust Oxidative Biocatalysts for the conversion and production of alcohols and our interest is to apply the combined knowledge and the analytical toolbox for the analysis of active pharmaceutical ingredients (APIs) to characterize oxidizing enzymes from the project partners.

    This project is carried out within acib in close collaboration with Prof. Anton Glieder (TUGraz) and DSM Chemical Technology R&D BV.

    The research for this work has received funding from the European Union (EU) project ROBOX (grant agreement n° 635734) under  EU’s Horizon 2020 Programme  Research and Innovation actions H2020-LEIT BIO-2014-1

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Past Projects

  • December 2012 - November 2015 - FP7 Project KYROBIO: Grant agreement no: 289646



    The project title is The discovery, development and demonstration of biocatalysts for use in the industrial synthesis of chiral chemicals and our interest was to identify a new class of hydroxynitrile lyases. We found a very promising enzyme in the fern Davallia teyermanii and characterized it in detail.

    This project was carried out within acib in close collaboration with Prof. Karl Gruber (UniGraz), Prof. Ruth Birner-Grünberger (ZMF and Omics Centre) and Dr. Gerhard Thallinger (TUGraz).


  • April 2014 - June 2015 - acib Project 84.011: Reverse nitrilase: From ammonium plus carboxylic acid to amine precursors

    The aim was to explore the substrate scope of enzymes that can convert a carboxylic acid to a nitrile.


  • January 2010 - December 2014 - acib Project 1.4-2: Enzymatic API Modification; Non-CYP drug metabolizing enzymes

    The aim was to make human drug metabolizing enzymes available in microbial hosts so that it is possible to prepare authentic human drug metabolites on the preparative scale.

    This project is carried out within acib in close collaboration with Novartis and F.Hoffmann-LaRoche.


  • January 2010 - December 2014 - acib Project 1.5-1: Amines and Peptides; Generation of efficient production strains for chiral amine synthesis

    The aim was to provide robust enzymes and yeast strains for the production of chiral amines.

    This project is carried out within acib in close collaboration with Prof. Anton Glieder, Dr. Andrea Camattari (TUGraz) and Ingenza Ltd.


  • January 2010 - December 2014 - acib Projects 1.2 and 4.12: Enzymatic Nitrile Reduction

    In this project we cloned, mutagenized and provided nitrile reductase enzymes to project partners with the aim to broaden its substrate specificity.

    This project is carried out within acib in close collaboration with Prof. Norbert Klempier, Prof. Bernd Nidetzky (TUGraz) and Prof. Karl Gruber (UniGraz).


  • January 2010 - June 2013 - acib Project 4.2-3: Enzyme engineering for complex reactions; Identification, expression and engineering of new complex microbial oxidoreductases

    The aim was to find enzymes for the reduction or oxidation of industrially relevant compounds.

    This project is carried out within acib in close collaboration with Prof. Helmut Schwab, Prof. Peter Macheroux (TUGraz) and Lonza AG.


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Current Theses


  • Melissa Horvat
    PhD thesis

    Working title:Carboxylic Acid Reductases for Enzymatic Reduction of organic Acids to Aldehydes

  • Lukas Kargl
    Bachelor thesis

    Working title:Kinetic characterization of CYP505 expressing biocatalysts
  • Daniel Schwendenwein
    PhD thesis
    Working title:Catalytic cascades for aldehyde synthesis in vivo

  • Philippe Oberhemm
    Master thesis
    Working title: Characterization of new carboxylate reductase enzymes

  • Holly Stolterfoht
    Master thesis
    Working title: Characterization and engineering of Carboxylate reductase

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Completed Theses


  • Katharina Dokulil
    Master Thesis
    Cloning, expression and characterization of 7-cyano-7-deazaguanine synthases
    May 2016
  • Thorsten Bachler
    Bachelor Thesis

    Human enzymes for organic synthesis

    April 2016

  • Elisa Lanfranchi
    PhD Thesis
    New Hydroxynitrile Lyase from white Rabbit's Foot Fern: The route from discovery to perspectives for applications
    Nov 2015
  • Kamila Napora-Wijata
    PhD Thesis
    Identification and expression of new microbial oxidoreductases
    February 2014
  • Thomas Prossliner
    Bachelor Thesis
    Characterization of a short-chain dehydrogenase/reductase from unconventional yeast Yarrowia lipolytica
    May 2013

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Partners


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Selected Publications

2017
Thomas Bayer, Thomas Wiesinger, Sofia Milker, Margit Winkler, Marko D. Mihovilovic and Florian Rudroff IN VIVO SYNTHESIS OF POLYHYDROXYLATED COMPOUNDS FROM A ‘HIDDEN RESERVOIR’ OF TOXIC ALDEHYDE SPECIES Show publication in PURE
Elisa Lanfranchi, Tea Pavkov-Keller, Eva-Maria Koehler, Matthias Diepold, Kerstin Steiner, Barbara Darnhofer, Jürgen Hartler, Tom Van Den Bergh, Henk Jan Joosten, Mandana Gruber-Khadjawi, Gerhard Thallinger, Ruth Birner-Gruenberger, Karl Gruber, Margit Winkler and Anton Glieder Enzyme discovery beyond homology: a unique hydroxynitrile lyase in the Bet v1 superfamily Show publication in PURE
Holly Stolterfoht, Daniel Schwendenwein, Christoph Wilhelm Sensen, Florian Rudroff and Margit Winkler Four distinct types of E.C. 1.2.1.30 enzymes can catalyze the reduction of carboxylic acids to aldehydes Show publication in PURE
2016
Margit Winkler and Christoph K. Winkler TRAMETES VERSICOLOR CARBOXYLATE REDUCTASE UNCOVERED Show publication in TUGRAZonline
Filippo Fiorentini, Martina Geier, Claudia Binda, Margit Winkler, Kurt Faber, Mélanie Hall and Andrea Matteiv Biocatalytic Characterization of Human FMO5: Unearthing Baeyer-Villiger Reactions in Humans Show publication in TUGRAZonline
Daniel Schwendenwein, Giuseppe Fiume, Hansjörg Weber, Florian Rudroff and Margit Winkler Selective Enzymatic Transformation to Aldehydes in vivo by Fungal Carboxylate Reductase from Neurospora crassa Show publication in PURE
Katrin Weinhandl, Melanie Ballach, Margit Winkler, Ahmad Mudassar, Anton Glieder, Ruth Birner-Gruenberger, Ian Fotheringham and Franck Escalettes Pichia pastoris mutants as host strains for efficient secretion of recombinant Branched Chain Aminotransferase (BCAT) Show publication in PURE
Marcia Ferreira Antunes, Fabian K. Eggimann, Matthias Kittelmann, Stephan Lütz, Steven P. Hanlon, Beat Wirz, Thorsten Bachler and Margit Winkler Human xanthine oxidase recombinant in E. coli: A whole cell catalyst for preparative drug metabolite synthesis Show publication in PURE
Margit Winkler and Christoph K. Winkler TRAMETES VERSICOLOR CARBOXYLATE REDUCTASE UNCOVERED Show publication in PURE
Filippo Fiorentini, Martina Geier, Claudia Binda, Margit Winkler, Kurt Faber, Mélanie Hall and Andrea Matteiv Biocatalytic Characterization of Human FMO5: Unearthing Baeyer-Villiger Reactions in Humans Show publication in PURE
2015
Margit Winkler Drug Metabolite Synthesis with Human Enzymes Show publication in TUGRAZonline
Margit Winkler, Katharina Dokulil, Hansjörg Weber, Tea Pavkov-Keller and Birgit Wilding The Nitrile forming Enzyme 7-Cyano-7-deazaguanine Synthase from Geobacillus kaustophilus: A reverse Nitrilase? Show publication in TUGRAZonline
Martina Geier, Thorsten Bachler, Steven P Hanlon, Fabian K Eggimann, Matthias Kittelmann, Hansjörg Weber, Stephan Lütz, Beat Wirz and Margit Winkler Human FMO2-based microbial whole-cell catalysts for drug metabolite synthesis Show publication in TUGRAZonline
Elisa Lanfranchi, Eva-Maria Köhler, Barbara Darnhofer, Kerstin Steiner, Ruth Birner-Grünberger, Anton Glieder and Margit Winkler Bioprospecting for Hydroxynitrile Lyases by Blue Native PAGE Coupled HCN Detection Show publication in TUGRAZonline
Corinna Odar, Margit Winkler and Birgit Wiltschi FLUORINATED AMINO ACIDS – A RARITY IN NATURE YET A PROSPECT FOR PROTEIN ENGINEERING Show publication in TUGRAZonline
Elisa Lanfranchi, Eva-Maria Köhler, Barbara Darnhofer, Kerstin Steiner, Ruth Birner-Grünberger, Anton Glieder and Margit Winkler Bioprospecting for Hydroxynitrile Lyases by Blue Native PAGE Coupled HCN Detection Show publication in PURE
Margit Winkler Drug Metabolite Synthesis with Human Enzymes Show publication in PURE
Corinna Odar, Margit Winkler and Birgit Wiltschi FLUORINATED AMINO ACIDS – A RARITY IN NATURE YET A PROSPECT FOR PROTEIN ENGINEERING Show publication in PURE
Martina Geier, Thorsten Bachler, Steven P Hanlon, Fabian K Eggimann, Matthias Kittelmann, Hansjörg Weber, Stephan Lütz, Beat Wirz and Margit Winkler Human FMO2-based microbial whole-cell catalysts for drug metabolite synthesis Show publication in PURE
Margit Winkler, Katharina Dokulil, Hansjörg Weber, Tea Pavkov-Keller and Birgit Wilding The Nitrile forming Enzyme 7-Cyano-7-deazaguanine Synthase from Geobacillus kaustophilus: A reverse Nitrilase? Show publication in PURE
2014
Katrin Weinhandl, Margit Winkler, Anton Glieder and Andrea Camattari Extracellular transaminases for biocatalysis New BiotechnologyS198 - S198 Show publication in TUGRAZonline
Margit Winkler, Martina Geier, Thorsten Bachler, Steven P Hanlon, Matthias Kittelmann, Anton Glieder, Stephan Lütz and Beat Wirz Human flavin monooxygenase 2: Heterologous expression in E. coli and API modificationNew BiotechnologyS82 - S82 Show publication in TUGRAZonline
Margit Winkler, Kamilla Napora-Wijata, Birgit Wilding and Norbert Klempier Unconventional Enzymatic Reductions: Nitriles and Carboxylates as SubstratesNew BiotechnologyS1 - S2 Show publication in TUGRAZonline
Steven P. Hanlon, Matthias Kittelmann and Margit Winkler SEMI-PREPARATIVE SCALE DRUG METABOLITE SYNTHESIS WITH HUMAN FLAVIN MONOOXYGENASESPractical Methods in Biocatalysis and Biotransformationsn/a - n/a Show publication in TUGRAZonline
Diogo Rodrigues, Matthias Kittelmann, Fabian Eggimann, Thorsten Bachler, Sandra Abad, Andrea Camattari, Anton Glieder, Margit Winkler and Stephan Lütz PRODUCTION OF RECOMBINANT HUMAN ALDEHYDE OXIDASE IN E. COLI AND OPTIMIZATION OF ITS APPLICATION FOR PREPARATIVE SYNTHESIS OF OXIDIZED DRUG METABOLITES Show publication in TUGRAZonline
Kamila Napora, Gernot Strohmeier and Margit Winkler BIOCATALYTIC REDUCTION OF CARBOXYLIC ACIDS Show publication in TUGRAZonline
Katrin Weinhandl, Margit Winkler, Anton Glieder and Andrea Camattari Carbon source dependent promoters in yeasts Show publication in TUGRAZonline
Margit Winkler, Martina Geier, Thorsten Bachler, Steven P Hanlon, Matthias Kittelmann, Anton Glieder, Stephan Lütz and Beat Wirz Human flavin monooxygenase 2: Heterologous expression in E. coli and API modificationNew BiotechnologyS82-S82 Show publication in PURE
Margit Winkler, Kamilla Napora-Wijata, Birgit Wilding and Norbert Klempier Unconventional Enzymatic Reductions: Nitriles and Carboxylates as SubstratesNew BiotechnologyS1-S2 Show publication in PURE
Katrin Weinhandl, Margit Winkler, Anton Glieder and Andrea Camattari Extracellular transaminases for biocatalysisNew BiotechnologyS198-S198 Show publication in PURE
Katrin Weinhandl, Margit Winkler, Anton Glieder and Andrea Camattari Carbon source dependent promoters in yeasts Show publication in PURE
Kamila Napora, Gernot Strohmeier and Margit Winkler BIOCATALYTIC REDUCTION OF CARBOXYLIC ACIDS Show publication in PURE
Diogo Rodrigues, Matthias Kittelmann, Fabian Eggimann, Thorsten Bachler, Sandra Abad, Andrea Camattari, Anton Glieder, Margit Winkler and Stephan Lütz PRODUCTION OF RECOMBINANT HUMAN ALDEHYDE OXIDASE IN E. COLI AND OPTIMIZATION OF ITS APPLICATION FOR PREPARATIVE SYNTHESIS OF OXIDIZED DRUG METABOLITES Show publication in PURE
Steven P. Hanlon, Matthias Kittelmann and Margit Winkler SEMI-PREPARATIVE SCALE DRUG METABOLITE SYNTHESIS WITH HUMAN FLAVIN MONOOXYGENASESPractical Methods in Biocatalysis and Biotransformations135-139 Show publication in PURE
2013
Elisa Lanfranchi, Kerstin Steiner, Anton Glieder, Ivan Hajnal, Roger A. Sheldon, Sander van Pelt and Margit Winkler RECENT DEVELOPMENTS IN HYDROXYNITRILE LYASES FOR INDUSTRIAL BIOTECHNOLOGY Show publication in TUGRAZonline
Birgit Wilding, Margit Winkler, Barbara Petschacher, Regina Kratzer, Sigrid Egger, Georg Steinkellner, Andrzej Franciszek Lyskowski, Bernd Nidetzky, Karl Gruber and Norbert Klempier Targeting the Substrate Binding Site of E. coli Nitrile Reductase QueF by Modeling, Substrate and Enzyme Engineering Show publication in TUGRAZonline
Kamila Napora, Tanja Wrodnigg, Patrick Kosmus, Martin Thonhofer, Karen Robins and Margit Winkler YARROWIA LIPOLYTICA DEHYDROGENASE/ REDUCTASE: AN ENZYME TOLERANT FOR LIPOPHILIC COMPOUNDS AND CARBOHYDRATE SUBSTRATES Show publication in TUGRAZonline
Elisa Lanfranchi, Kerstin Steiner, Anton Glieder, Ivan Hajnal, Roger A. Sheldon, Sander van Pelt and Margit Winkler RECENT DEVELOPMENTS IN HYDROXYNITRILE LYASES FOR INDUSTRIAL BIOTECHNOLOGY Show publication in PURE
Birgit Wilding, Margit Winkler, Barbara Petschacher, Regina Kratzer, Sigrid Egger, Georg Steinkellner, Andrzej Franciszek Lyskowski, Bernd Nidetzky, Karl Gruber and Norbert Klempier Targeting the Substrate Binding Site of E. coli Nitrile Reductase QueF by Modeling, Substrate and Enzyme Engineering Show publication in PURE
Kamila Napora, Tanja Wrodnigg, Patrick Kosmus, Martin Thonhofer, Karen Robins and Margit Winkler YARROWIA LIPOLYTICA DEHYDROGENASE/ REDUCTASE: AN ENZYME TOLERANT FOR LIPOPHILIC COMPOUNDS AND CARBOHYDRATE SUBSTRATES Show publication in PURE
2012
Margit Winkler and Andrea Camattari Recombinant Human Flavin Monooxygenases: Expression and Moclobemide N-oxide Synthesis on Semi-Preparative ScaleNew BiotechnologyS236 - S237 Show publication in TUGRAZonline
Katrin Weinhandl, Margit Winkler, Anton Glieder and Andrea Camattari A NOVEL MULTI-ENZYMATIC ASSAY FOR TRANSAMINASE ACTIVITY: HIGH THROUGHPUT SCREENING APPLICATIONS IN PICHIA PASTORIS Show publication in TUGRAZonline
Birgit Wilding, Margit Winkler, Regina Kratzer, Barbara Petschacher, Anton Glieder and Norbert Klempier NITRILE REDUCTASE FROM GEOBACILLUS KAUSTOPHILUS: A POTENTIAL CATALYST FOR A NEW NITRILE BIOTRANSFORMATION REACTION Show publication in TUGRAZonline
Margit Winkler Organ Function in Shake Flasks: Recombinant Human Enzymes for the Pharmaceutical Industry Show publication in TUGRAZonline
Steven P. Hanlon, Andrea Camattari, Sandra Abad, Anton Glieder, Beat Wirz, Matthias Kittelmann, Stephan Lütz and Margit Winkler EXPRESSION OF RECOMBINANT HUMAN FLAVIN MONOOXYGENASE AND MOCLOBEMIDE N-OXIDE SYNTHESIS ON MULTI-MG SCALE Show publication in TUGRAZonline
Margit Winkler, Anton Glieder and Kerstin Steiner C–X Bond Formation: Hydroxynitrile lyases: From Nature to ApplicationComprehensive Chirality350 - 371 Show publication in TUGRAZonline
Margit Winkler and David O'Hagan Synthesis of Fluorinated Neurotransmitter AnaloguesFluorine in Pharmaceutical and Medicinal Chemistry : From Biophysical Aspects to Clinical Applications299 - 331 Show publication in TUGRAZonline
Margit Winkler Organ Function in Shake Flasks: Recombinant Human Enzymes for the Pharmaceutical Industry Show publication in PURE
Katrin Weinhandl, Margit Winkler, Anton Glieder and Andrea Camattari A NOVEL MULTI-ENZYMATIC ASSAY FOR TRANSAMINASE ACTIVITY: HIGH THROUGHPUT SCREENING APPLICATIONS IN PICHIA PASTORIS Show publication in PURE
Birgit Wilding, Margit Winkler, Regina Kratzer, Barbara Petschacher, Anton Glieder and Norbert Klempier NITRILE REDUCTASE FROM GEOBACILLUS KAUSTOPHILUS: A POTENTIAL CATALYST FOR A NEW NITRILE BIOTRANSFORMATION REACTION Show publication in PURE
Margit Winkler, Anton Glieder and Kerstin Steiner C–X Bond Formation: Hydroxynitrile lyases: From Nature to ApplicationComprehensive Chirality350-371 Show publication in PURE
Steven P. Hanlon, Andrea Camattari, Sandra Abad, Anton Glieder, Beat Wirz, Matthias Kittelmann, Stephan Lütz and Margit Winkler Expression of recombinant human flavin monooxygenase and moclobemide-N-oxide synthesis on multi-mg scale Show publication in PURE
Margit Winkler and David O'Hagan Synthesis of Fluorinated Neurotransmitter AnaloguesFluorine in Pharmaceutical and Medicinal Chemistry : From Biophysical Aspects to Clinical Applications299-331 Show publication in PURE
Margit Winkler and Andrea Camattari Recombinant Human Flavin Monooxygenases: Expression and Moclobemide N-oxide Synthesis on Semi-Preparative ScaleNew BiotechnologyS236-S237 Show publication in PURE
2011
Sandra Abad, Jozef Nahalka, Margit Winkler, Gabriele Bergler, Robert Speight, Anton Glieder and Bernd Nidetzky HIGH LEVEL EXPRESSION OF RHODOTORULA GRACILIS D-AMINO ACID OXIDASE IN PICHIA PASTORIS Show publication in TUGRAZonline
Matthias Schittmayer, Anton Glieder, Michael Karl Uhl, Andreas Winkler, Simone Zach, Jörg Schrittwieser, Wolfgang Kroutil, Peter Macheroux, Karl Gruber, Spiros Kambourakis, J. David Rozzell and Margit Winkler Old Yellow Enzyme catalyzed dehydrogenation of saturated ketones Show publication in TUGRAZonline
Sandra Abad, Jozef Nahalka, Margit Winkler, Gabriele Bergler, Robert Speight, Anton Glieder and Bernd Nidetzky HIGH LEVEL EXPRESSION OF RHODOTORULA GRACILIS D-AMINO ACID OXIDASE IN PICHIA PASTORIS Show publication in PURE
Matthias Schittmayer, Anton Glieder, Michael Karl Uhl, Andreas Winkler, Simone Zach, Jörg Schrittwieser, Wolfgang Kroutil, Peter Macheroux, Karl Gruber, Spiros Kambourakis, J. David Rozzell and Margit Winkler Old Yellow Enzyme catalyzed dehydrogenation of saturated ketones Show publication in PURE
2010
Ulrike Schreiner, Bianca Hecher, Sascha Obrowsky, Kerstin Waich, Norbert Klempier, Georg Steinkellner , Karl Gruber, J. David Rozzell, Anton Glieder and Margit Winkler Directed evolution of Alcaligenes faecalis nitrilase Show publication in TUGRAZonline
Daniel E Torres Pazmiño, Margit Winkler, Anton Glieder and Marco W Fraaije Monooxygenases as biocatalysts: classification, mechanistic aspects and biotechnological applications Show publication in TUGRAZonline
Ulrike Schreiner, Georg Steinkellner, Anton Glieder, J. David Rozzell and Margit Winkler IMPROVED FITNESS OF ARABIDOPSIS THALIANA NITRILASE 2 Show publication in TUGRAZonline
Ulrike Schreiner, Bianca Hecher, Sascha Obrowsky, Kerstin Waich, Norbert Klempier, Georg Steinkellner, Karl Gruber, J. David Rozzell, Anton Glieder and Margit Winkler Directed evolution of Alcaligenes faecalis nitrilase Show publication in PURE
Daniel E Torres Pazmiño, Margit Winkler, Anton Glieder and Marco W Fraaije Monooxygenases as biocatalysts: classification, mechanistic aspects and biotechnological applications Show publication in PURE
Ulrike Schreiner, Georg Steinkellner, Anton Glieder, J. David Rozzell and Margit Winkler IMPROVED FITNESS OF ARABIDOPSIS THALIANA NITRILASE 2 Show publication in PURE

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