the winkler lab

Biocatalysis and Biotechnology for Chemical Synthesis

We are fascinated by the power of natural catalysts (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 our 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.

SELECTED PROJECTS

A new chemoenzymatic route from carboxylic acids to nitriles

Nitriles are important compounds in chemistry. Citronellyl nitrile, as an example, has a pleasant lemon-like smell, and is widely used in home-care products. The classical way to prepare nitriles is by the reaction of a precursor molecule with cyanide. However, cyanide is highly toxic. In Agatha Christie´s murder stories, the murderer takes the victim’s breath with “sparkling cyanide”: a bit of the inconspicuous white powder potassium cyanide mixed into sparkling wine. To avoid the use of cyanide in chemical syntheses, we propose to apply enzymes. They are non-toxic and typically operate at ecologically benign conditions.

 

toc-proof

The aim of this project is, to combine two enzymes (carboxylate reductase and aldoxime dehydratase) and a simple chemical reaction to sequentially convert carboxylic acids to aldehydes, oximes and finally to produce nitriles. Carboxylate reductase enzymes can selectively make an aldehyde from a carboxylic acid using cofactors as the energy source. These cofactors, however, are too expensive to be added to the reaction and therefore, we will use microorganisms, which easily produce these cofactors from sugar and air. The product of this first step– the aldehyde – is very reactive and will be trapped immediately in a chemical reaction as an oxime. This compound is then recognized by aldoxime dehydratase as a substrate. Aldoxime dehydratases are remarkable enzymes, because of their unique reaction mechanism, the removal of water from the oxime in aqueous solution. This is a mild alternative to the harsh chemical equivalent to this reaction which proceeds at high temperature and requires high energy input.

One of the key outcomes of this project will be a better understanding of the interplay between enzymes and chemicals within the complex system of a living microorganism. This knowledge allows us to spot bottlenecks in the reaction cascade and to find solutions how to overcome them. Our results will provide valuable input for other cascade reactions with aldehyde intermediates. From genome sequencing projects we know that hundreds of genes encoding aldoxime dehydratases are present in bacteria and fungi. However only a handful of these enzymes have been studied. Therefore, one of our goals is to ”revive” slumbering aldoxime dehydratase genes. An important additional outcome will be a deeper knowledge of the so far rarely studied enzyme class of aldoxime dehydratases and new variants of these highly interesting biocatalysts.

This Project is carried out in close collaboration with the Czech Academy of Sciences (Ludmila Martínková, Miroslav Patek) and the TU Wien (Florian Rudroff)

This project received funding from the Austrian Science Fund FWF (I 04607). 

Catalytic cascades for selective aldehyde syntheses in vivo

Many fragrances that we know from daily life share the same chemical functional group – the aldehyde. The most prominent example is probably vanillin, which is used in a huge number of food products, cosmetics and perfumes but there are also others like cinnamaldehyde (which gives the smell to cinnamon) and derivatives thereof. As many of these fragrances are difficult to obtain in sufficient amounts from natural sources, there is a lot of interest to find ways how to prepare them. Carboxylic acids could be promising precursors, but it is very difficult to reduce them selectively to aldehydes. Chemistry has some possibilities to do that, but they all suffer from drawbacks that enzymatic methods can circumvent. Enzymes are non-toxic and typically operate at ecologically benign conditions. So called carboxylate reductase enzymes can selectively make an aldehyde from an acid at the expense of cofactors.

These cofactors, however, are too expensive to be added to a reaction and therefore, we will use microorganisms, which produce these cofactors from sugar and air. The microorganism E. coli will be engineered such that they are able to catalyze a cascade reaction to fragrances like for example TropionalTM, which has an ‘ocean – like’ smell, from precursors that were synthesized with catalytic chemical methods. Ultimately, the aim is to integrate the chemical step into the biotransformation step and run the reaction in a chemoenzymatic one pot fashion.

This Project wa carried out in close collaboration with the TU Wien (Florian Rudroff).

This project received funding from the Austrian Science Fund FWF (P 28477-B21). 

Current team

Priv.-Doz. Dr. Margit Winkler
Principal Investigator

TU Graz, IMBT Team Pics

Valentina Jurkas
PostDoc

Lukas Schober
PhD student

Karin Reicher
Researcher

Holly Stolterfoht-Stock
Master student & Researcher

Hannah Breuer
Student coworker, Master student

Svitlana Münzer
Support

Alexandra Deibuk
Master student

Dominic Goj
Master student, student co-worker

Past team-members

rhdr

A.Pleh
Student coworker

Cham Hashem
Researcher

N.N.
Bachelor student

N.N.
PhD guest student

Elena Pietrich
Technician

Melissa Horvat
PhD student & PostDoc

Stephanie Gerhofer
Bachelor student

Hana Dobiasová
PhD guest student

Tanja-Saskia Larch
Master student & Researcher

Lukas Pfeifenberger
Master student

Victoria Weilch
Master student

Stella Ebner
Master student

K. Gimpl
Researcher

Masethabela Maria Maphatsoe
Guest PhD student

M. Glänzer
Master student

S. Steinberger
Master student

dsc_2959

Viktorija Vidimce-Risteski
Researcher

David Gradischnig
Student co-worker & Bachelor student

Anna Schwarz
Master student

Daniel Schwendenwein
persistent identifier: PhD student

Birgit Grill
Researcher

U. Pogorevcnik
Student co-worker

Susanne Fritsche
Master student

Inge Eiteljörg
Staff Scientist

Thorsten Bachler
Researcher

B. Kerschbaumer
Master student

M. Willim
Master student

Giuseppe Fiume
Guest student

Katrin Weinhandl
Researcher

P. Oberhemm
Master student

Elisa Lanfranchi
PhD student

Kamila Napora-Wijata
PhD student

E.-M. Köhler
Master student

K. Dokulil
Master student

G. Offenmüller
Student-Coworker

H. Mandl
Technician

gruppenfoto-juli-2016

PUBLICATIONS

Publication Tabs

Ludmila Martínková, Michal Grulich, Miroslav Pátek, Barbora Křístková, Margit Winkler Bio-based valorization of lignin-derived phenolic compounds: a review Biomolecules, 2023, 13:717. DOI: 10.3390/biom13050717
Lukas Schober, Dobiašová, Valentina Jurkaš, Fabio Parmeggiani, Florian Rudroff, Margit Winkler Enzymatic reactions towards aldehydes: an overview Flavour and Fragrance Journal, 2023, 38, 221-42. DOI: 10.1002/ffj.3739
Katharina Ebner, Lukas J. Pfeifenberger, Veronika Schusterbauer, Claudia Rinnofner, Anton Glieder, Margit Winkler Discovery and heterologous expression of unspecific Peroxygenases Catalysts, 2023, 13:, 206. DOI: 10.3390/catal13010206
Margit Winkler, Melissa Horvat, Astrid Schiefer, Victoria Weilch, Florian Rudroff, Miroslav Patek, Ludmila Martinková Organic Acid to Nitrile: a chemoenzymatic three-step route, Advanced Synthesis and Catalysis, 2023,  365, 37-42. DOI:10.1002/adsc.202201053
Barbora Křístková, Robert Rädisch, Natalia Kulik, Melissa Horvat, Lenka Rucká, Michal Grulich, Florian Rudroff, Miroslav Pátek, Margit Winkler and Ludmila Martínková Scanning aldoxime dehydratase sequence space and characterization of a new aldoxime dehydratase OxdFv from Fusarium vanettenii,Enz. Microb. Technol., 2023, 164: 110187. DOI: 110.1016/j.enzmictec.2022.110187
Chiam Hashem, Julius Hochrinner, Claudia Rinnofner, Moritz B. Bürgler, Harald Pichler, Margit Winkler From Linoleic acid to hexanal and hexanol by whole cell catalysis with a lipoxygenase, hydroperoxide lyase and reductase cascade in Komagataella phaffii, Frontiers in Molecular Biosciences, 2022, 9:, 965315. DOI:10.3389/fmolb.2022.965315
Bastian Daniel, Chiam Hashem, Marlene Leithold, Theo Sagmeister, Adrian Tripp, Holly Stolterfoht-Stock, Julia Messenlehner, Ronan Keegan, Christoph K. Winkler, Jonathan G. Ling, Sabry H.H. Younes, Gustav Oberdorfer, Farah D.A. Bakar, Karl Gruber, Tea Pavkov-Keller and Margit Winkler Structure of the Reductase Domain of a fungal Carboxylic Acid Reductase and its Substrate Scope in Thioester and Aldehyde Reduction, ACS Catalysis, 2022, 12, 15668-15674. DOI:10.1021/acscatal.2c04426
Margit Winkler, Jonathan Guyang Ling Biocatalytic carboxylate reduction – recent advances and new enzymes, ChemCatChem, 2022, 14, e202200441. DOI:10.1002/cctc.202200441
Robert Rädisch, Miroslav Patek, Barbora Křistková, Margit Winkler, Vladimír Křen, Ludmila Martinková Metabolism of Aldoximes and Nitriles in Plant-associated Bacteria and its Potential in Plant-Bacteria Interactions Microorganisms , 2022, 10: 549. DOI: 10.3390/microorganisms10030549
Masethabela Maria Maphatsoe, Chiam Hashem, Jonathan Guyang Ling, Melissa Horvat, Karl Rumbold, Farah Diba Abu Bakar, Margit Winkler Characterization and immobilization of Pycnoporus cinnabarinus carboxylic acid reductase, PcCAR2, J. Biotechnol., 2022, 345, 47-54. DOI:10.1016/j.jbiotec.2021.12.010
Melissa Horvat, Victoria Weilch, Robert Rädisch, Sebastian Hecko, Astrid Schiefer, Florian Rudroff, Birgit Wilding, Norbert Klempier, Miroslav Patek, Ludmila Martinková, Margit Winkler Chemoenzymatic one-pot reaction from carboxylic acid to nitrile via oxime Catalysis Science & Technology , 2022, 12, 62-66. DOI: 10.1039/D1CY01694F
Birgit Grill, Melissa Horvat, Helmut Schwab, Ralf Gross, Kai Donsbach, Margit Winkler Gordonia hydrophobica Nitrile Hydratase for Amide preparation from nitriles Catalysts, 2021, 11, 1287. DOI: 10.3390/catal11111287
Daniel Schwendenwein, Anna K. Ressmann, Marcello Entner, Viktor Savic, Margit Winkler, Florian Rudroff Chemo-Enzymatic Cascade for the Generation of Fragrance Aldehydes Catalysts, 2021, 11, 932. DOI: 10.3390/catal11080932
Hot Paper Sebastian Arndt, Birgit Grill, Helmut Schwab, Georg Steinkellner, Urška Pogorevčnik, Dominik Weis, Alexander M. Nauth, Karl Gruber, Till Opatz, Kai Donsbach, Siegfried R. Waldvogel, Margit Winkler The sustainable synthesis of levetiracetam by an enzymatic dynamic kinetic resolution and an ex-cell anodic oxidation Green Chemistry, 2021, 23, 388-395. DOI: 10.1039/D0GC03358H
Anna Schwarz, Sebastian Hecko, Florian Rudroff, Jeffrey Kohrt, Roger M. Howard, Margit Winkler, Cell-free in vitro reduction of carboxylates to aldehydes: with crude enzyme preparations to a key pharmaceutical building block Biotechnology Journal, 2020,  15, 2000315. DOI: 10.1002/biot.202000315
Melissa Horvat, Tanja-Saskia Larch, Florian Rudroff, Margit Winkler Amino benzamidoxime (ABAO) based assay to identify efficient aldehyde-producing Pichia pastoris clones, Advanced Synthesis and Catalysis, 2020,  362(21), 4673-4679. DOI:10.1002/adsc.202000558
Yating Hu, Zhiwei Zhu, David Gradischnig, Margit Winkler, Jens Nielsen, Verena Siewers Engineering carboxylic acid reductase for selective synthesis of medium-chain fatty alcohols in yeast PNAS2020,  117, 22974-22983. DOI: 10.1073/pnas.2010521117
Chiam Hashem, Holly Stolterfoht, Claudia Rinnofner, Stefan Steinberger, Margit Winkler, Harald Pichler Secretion of Pseudomonas aeruginosa lipoxygenase by Pichia pastoris upon glycerol feed Biotechnology Journal, 2020,  15, 2000089. DOI: 10.1002/biot.202000089
Frederick Mortzfeld, Chiam Hashem, Kvetoslava Vrankova, Margit Winkler, Florian Rudroff Pyrazines – valuable flavour & fragrance compounds: Biocatalytic synthesis and industrial applications. Biotechnology Journal, 2020,  15, 2000064. DOI: 10.1002/biot.202000064
Melissa Horvat, Margit Winkler In vivo reduction of medium- to long-chain fatty acids by CAR enzymes: limitations and solutions, ChemCatChem, 2020,  12, 5076-5090. DOI:10.1002/cctc.202000895
Birgit Grill, Maximilian Glänzer, Helmut Schwab, Kerstin Steiner, Daniel Pienaar, Dean Brady, Kai Donsbach, Margit Winkler Functional expression and characterization of a panel of cobalt and iron dependent nitrile hydratases, Molecules, 2020,  25, 2521. DOI:10.3390/molecules25112521
Melissa Horvat, Giuseppe Fiume, Susanne Fritsche, Margit Winkler Discovery of carboxylic acid reductase (CAR) from Thermothelomyces thermophila and its evaluation for vanillin synthesis, J. Biotechnol., 2019,  304, 44-51. DOI:10.1016/j.jbiotec.2019.08.007
Melissa Horvat, Susanne Fritsche, Robert Kourist, Margit Winkler Characterization of type IV carboxylate reductases (CARs) for whole cell mediated preparation of 3-hydroxytyrosol, ChemCatChem, 2019,  11, 4171-4181. DOI:10.1002/cctc.201900333
Daniel Schwendenwein, Anna K. Ressmann, Mark Dörr, Matthias Höhne, Marko D. Mihovilovic, Uwe T. Bornscheuer, Florian Rudroff, Margit Winkler Random Mutagenesis Driven Improvement of Carboxylate Reductase Activity by a Substrate Independent High-Throughput Assay in Living Cells, Adv. Synth. Catal., 2019,  361, 2544-2549. DOI:10.1002/adsc.201900155
Anna K. Ressmann, Daniel Schwendenwein, Simon Leonhartsberger, Marko D. Mihovilovic, Uwe T. Bornscheuer, Margit Winkler, Florian Rudroff Substrate Independent High-Throughput Assay for the Quantification of Aldehydes, Adv. Synth. Catal. 2019,  361, 2538-2543. DOI:10.1002/adsc.201900154
Gernot Strohmeier, Inge C. Eiteljörg, Anna Schwarz, Margit Winkler Enzymatic one step reduction of carboxylate to aldehyde with cell free regeneration of ATP and NADPH, Chem. Eur. J., 2019,  25, 6119-6123. DOI:10.1002/chem.201901147
Claudia Rinnofner, Bianca Kerschbaumer, Hansjörg Weber, Anton Glieder, Margit Winkler Cytochrome P450 mediated hydroxylation of ibuprofen using Pichia pastoris as biocatalyst, Biocatal. Agricult. Biotechnol., 2019, 525-528. DOI:10.1016/j.bcab.2018.12.022
Anna K. Migglautsch, Melissa Willim, Bettina Schweda, Anton Glieder, Rolf Breinbauer, Margit Winkler Aliphatic hydroxylation and epoxidation of capsaicin by cytochrome P450 CYP505X, Tetrahedron, 2018, 74, 6199-6204. DOI:10.1016/j.tet.2018.08.049
Margit Winkler, Martina Geier, Steven P. Hanlon, Bernd Nidetzky, Anton Glieder Human Enzymes for Organic Synthesis, Angew. Chem. Intl. Ed., 2018, 57, 13406-13423. DOI:10.1002/anie.201800678
Holly Stolterfoht, Georg Steinkellner, Daniel Schwendenwein, Tea Pavkov-Keller, Karl Gruber, Margit Winkler Identification of key residues of Carboxylate reductases, Front. Microbiol., 2018, 9: 250. DOI:10.3389/fmicb.2018.00250
Elisa Lanfranchi, Birgit Grill, Zainab Raghoebar, Sander Van Pelt, Roger A. Sheldon, Kerstin Steiner, Anton Glieder, Margit Winkler Production of hydroxynitrile lyase from D. tyermannii in Komagataella phaffii and its immobilization to generate a robust biocatalyst, ChemBioChem, 2018, 19 , 312-316. DOI:10.1002/cbic.201700419
 Margit Winkler Carboxylic acid Reductse Enzymes (CARs) Current Opinion in Chemical Biology, 2018, 43, 23-29. DOI:10.1016/j.cbpa.2017.10.006
Elisa Lanfranchi, Tea Pavkov-Keller, Eva-Maria Koehler, Matthias Diepold, Kerstin Steiner, Barbara Darnhofer, Juergen Hartler, Tom Van Den Bergh, Henk-Jan Joosten, Mandana Gruber-Khadjawi, Gerhard G. Thallinger, Ruth Birner-Gruenberger, Karl Gruber, Margit Winkler, Anton Glieder Enzyme discovery beyond homology: a unique hydroxynitrile lyase in the bet v1 superfamily Sci. Reports, 2017, 9:46738. DOI:10.1038/srep46738
 Thomas Bayer, Thomas Wiesinger, Sofia Milker, Margit Winkler, Marko D. Mihovilovic, Florian Rudroff In vivo synthesis of polyhydroxylated compounds from a 'hidden reservoir' of toxic aldehyde species, ChemCatChem, 2017, 9, 2919-2923. DOI: 10.1002/cctc.201700469
Holly Stolterfoht, Daniel Schwendenwein, Christoph W. Sensen, Florian Rudroff, Margit Winkler Four distinct types of E.C. 1.2.1.30 enzymes can catalyze the reduction of carboxylic acids to aldehydes, J. Biotechnol., 2017, 257, 222-232. DOI:10.1016/j.jbiotec.2017.02.014
Daniel Schwendenwein, Giuseppe Fiume, Hansjörg Weber, Florian Rudroff, Margit Winkler Selective enzymatic transformation to aldehydes in vivo by fungal carboxylate reductase from Neurospora crassa, Adv. Syn. Catal., 2016, 358, 3414-3421. DOI: 10.1002/adsc.201600914
Márcia Ferreira Antunes, Fabian K. Eggimann, Matthias Kittelmann, Stephan Lütz, Steven P. Hanlon, Beat Wirz, Thorsten Bachler, Margit Winkler Human xanthin oxidase recombinant in E. coli: a whole cell catalyst for preparative drug metabolite synthesis, J. Biotechnol., 2016, 235, 3-10. DOI:10.1016/j.jbiotec.2016.03.045
Margit Winkler, Christoph K. Winkler Trametes versicolor carboxylate reductase uncovered, Chem. Monthly, 2016, 147, 575-578. DOI:10.1007/s00706-016-1676-z
Margit Winkler, Katharina Dokulil, Hansjörg Weber, Tea Pavkov-Keller, Birgit Wilding The nitrile forming enzyme 7-Cyano-7-Deazaguanine synthase from Geobacillus kaustophilus: a reverse nitrilase? ChemBioChem, 2015, 147, 2373-2378. DOI:10.1002/cbic.201500335
Martina Geier, Thorsten Bachler, Steven P. Hanlon, Fabian K. Eggimann, Matthias Kittelmann, Hansjörg Weber, Stephan Lütz, Beat Wirz, Margit Winkler Human FMO2-based microbial whole-cell catalysts for drug metabolite synthesis, Microbial cell factories, 2015, 14: 82. DOI:10.1186/s12934-015-0262-0
Elisa Lanfranchi, Eva-Maria Köhler, Barbara Darnhofer, Kerstin Steiner, Ruth Birner-Gruenberger, Anton Glieder, Margit Winkler Bioprospecting for hydroxynitrile lyases by blue native PAGE coupled HCN detection, Curr. Biotechnol. , 2015,  4 , 111-117. DOI: 10.2174/2211550104666150506225048
Kamila Napora-Wijata, Gernot A. Strohmeier, Margit Winkler Biocatalytic reduction of carboxylic acids, Biotechnol. J. , 2014,  9 , 822-843. DOI: 10.1002/biot.201400012
Kamila Napora-Wijata, Karen Robins, Antonio Osorio-Lozada, Margit Winkler Whole cell carboxylate reduction for the synthesis of 3-hydroxytyrosol, ChemCatChem , 2014,  6 , 1089-1095. DOI: 10.1002/cctc.201300913
Elisa Lanfranchi, Kerstin Steiner, Anton Glieder, Ivan Hajnal, Roger A. Sheldon, Sander van Pelt, Margit Winkler Recent developments in hydroxynitrile lyases for industrial biotechnology, Rec. Patents Biotechnol. , 2013,  7 , 197-206.
Kamila Napora-Wijata, Gernot A. Strohmeier, Manoj N. Sonavane, Manuela Avi, Karen Robins, Margit Winkler Enantiocomplementary Yarrowia lipolytica oxidoreductases: alcohol dehydrogenase 2 and short chain dehydrogenase/reductase, Biomolecules , 2013, 3, 449-460.
Kamila Napora, Tanja M. Wrodnigg, Patrick Kosmus, Martin Thonhofer, Karen Robins, Margit Winkler Yarrowia lipolytica dehydrogenase/reductase: an enzyme tolerant for lipophilic compounds and carbohydrate substrates, Bioorg. Med. Chem. Lett. , 2013, 23, 3393-3395.
Steven P. Hanlon, Andrea Camattari, Sandra Abad, Anton Glieder, Beat Wirz, Matthias Kittelmann, Stephan Lütz, Margit Winkler Expression of recombinant human flavin monooxygenase and moclobemide N-oxide synthesis on multi-mg scale, Chem. Commun. , 2012, 48, 6001-6003.
Matthias Schittmayer, Anton Glieder, Michael K. Uhl, Andreas Winkler, Simone Zach, Jörg H. Schrittwieser, Wolfgang Kroutil, Peter Macheroux, Karl Gruber, Spiros Kambourakis, J. David Rozzell, Margit Winkler Old yellow enzyme catalyzed dehydrogenation of saturated ketones, Adv. Syn. Catal. , 2011, 252, 268-274.
Ulrike Schreiner, Bianca Hecher, Sascha Obrowsky, Kerstin Waich, Norbert Klempier, Georg Steinkellner, Karl Gruber, J. David Rozzell, Anton Glieder, Margit Winkler Directed evolution of Alcaligenes faecalis nitrilase, Enz. Microb. Technol. , 2010, 47, 140-146.
Ulrike Schreiner, Georg Steinkellner, J. David Rozzell, Anton Glieder, Margit Winkler Improved fitness of Alcaligenes faecalis nitrilase 2, ChemCatChem , 2010, 2, 267-269.
Margit Winkler, Thomas Moraux, Hesham A. Khairy, Roderick H. Scott, Alexandra M. Z. Slawin, David O’Hagan Synthesis and vanilloid receptor (TRPV1) activity of the enantiomers of α-fluorinated capsaicin, ChemBioChem , 2009, 10, 823-828.
Margit Winkler, Norbert Klempier Enantioseparation of nonproteinogenic amino acids, Anal. Bioanal. Chem. 2009, 939, 1789-1796.
Margit Winkler, Ondřej Kaplan, Vojtěch Vejvoda, Norbert Klempier, Ludmila Martínková Biocatalytic application of nitrilases from Fusarium solani O1 Aspergillus niger K10, J. Mol.Catal. B: Enzym. 2009, 59, 243-247.
Margit Winkler, Juozas Domarkas, Lutz F. Schweiger, David O’Hagan Fluorinase-coupled base swaps: Synthesis of [18F]-5'-deoxy-5'-fluorouridines, Angew. Chem. Int. Ed. 2008, 47, 10141-10143.
Margit Winkler, Norbert Klempier, Hansjörg Weber, Katrin R. Grünwald, Michaela Flock, Alk Dransfeld Influence of relative configuration of disubstituted cyclopentanes and -hexanes on 13C shifts, Magn. Res. Chem. 2008, 46, 865-871.
Margit Winkler, Astrid-C. Knall, Martin R. Kulterer, Norbert Klempier Nitrilases catalyze key step to conformationally constrained GABA analogous γ-amino acids in high optical purity, J. Org. Chem. 2007, 72, 7423-7426.
Margit Winkler, Dorith Meischler, Norbert Klempier Nitrilase-catalyzed enantioselective synthesis of pyrrolidine- and piperidinecarboxylic acids, Adv. Syn. Catal. 2007, 349, 1475-1480.
Margit Winkler, Anton Glieder Norbert Klempier Enzyme stabilizer DTT catalyzes nitrilase analogue hydrolysis of nitriles, Chem. Commun. 2006, 12, 1298-1300.
Margit Winkler, Ludmila Martínková, Astrid C. Knall, Stefan Krahulec, Norbert Klempier Synthesis and microbial transformation of β-amino nitriles, Tetrahedron 2005, 61, 4249-4260.
Invited
 Margit Winkler (2023) Disruptive news about enzymatic carboxylic acid reduction, Novel Enzymes 2023, Mar 28-31, Greifswald, Germany
 Margit Winkler (2022) Invited Enzymatic Functional Group Transformations around aldehydes and nitriles, Symposium “New Trends in Organic Synthesis”, Nov 28, Milano, Italy
 Margit Winkler (2022) Enzyme assisted routes to flavor aldehydes and nitriles, Bioflavour 2022, Sep 20-22, Frankfurt, Germany
Margit Winkler (2022) Invited Expanding the toolbox for biocatalytic aldehyde synthesis Biocat 2022 – 10th International Congress on Biocatalysis, Aug.28-Sep 1, Hamburg, Germany
Margit Winkler (2021) Invited Recent advances in enzymatic reduction of carboxylic acids 3rd CCBIO Symposium, Jun. 10, online meeting
Margit Winkler, Florian Rudroff (2020) Reaction of aldehydes with aminobenzamidoxime: Scope and Applications as Analytical Tool ÖGMBT Life Science Tuesday, Dec. 15, online meeting
 Margit Winkler (2019) Towards flavors and fragrances with enzymes and microbes Center of Biotechnology and Functional Food, Universiti Kebangsaan Malaysia, Nov. 15, Bangi, Malaysia
 Margit Winkler (2019) Invited Plenary From fatty acids to flavors and fragrances with enzymes or microbes International Congress of the Malaysian Society for Microbiology 2019 (ICMSM2019), Nov. 11-14, Negeri Sembilan, Malaysia
 Margit Winkler (2019) Invited Enzymatische Reduktion von (Fett)Säuren, DGF Tagung 2019, Nov 7, Hamburg, Germany
 Margit Winkler (2019) In vivo – in vitro – in sequence: aldehyde formation by CARs, Biotrans 2019, Jul 7-11, Groningen, The Netherlands
 Margit Winkler (2019) Invited Keynote Lecture Harnessing carboxylate reductases to valorize carboxylates Biotech France 2019, Jun. 26, Paris, France
 Margit Winkler (2019) Mit Enzymen und Mikroben zu Bioflavours Vortrag aus der Sensorik Netzwerk Österreich Vortragsreihe 2019, Jun.4, Vienna, Austria
Margit Winkler, Karl Rumbold (2018) Plant hydroxynitrile lyase diversity and application for industrial biotechnology, Austria - South Africa S&T Cooperation Closing Workshop, Dec 12, Pretoria, South Africa
 Margit Winkler (2018) Invited Discovery, characterization and application of carboxylic acid reductases Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Nov. 6, Bangi, Malaysia
 Margit Winkler (2018) A greener route to aldehydes: enzymatic carboxylate reduction Bioflavour 2018, Sep 18-21, Frankfurt, Germany
 Margit Winkler (2018) Carboxylic acid reductase enzymes (CARs) 18TH European Congress on Biotechnology, Jul 1-4, Geneva, Switzerland
 Daniel Schwendenwein (2018) Heliotropin preparation by a carboxylate reductase in resting cells 19th DocDay, Feb. 9, Graz, Austria
 Margit Winkler (2017) Invited Keynote Lecture Discovery of new enzymes by homology and beyond International Congress of the Malaysian Society for Microbiology 2017 (ICMSM2017), Dec.6, Bandar Baru Bangi, Malaysia
 Daniel Schwendenwein (2017) Heliotropin preparation by a carboxylate reductase in resting cells School of Molecular and Cell Biology, University of the Witwatersrand, Nov.23, Johannesburg, South Africa
 Margit Winkler (2017) Enzymatic Functional Group Transformations for Industrial Biotechnology CATSA 2017, Nov.21, Sun City, South Africa
 Daniel Schwendenwein (2017) Heliotropin preparation by a carboxylate reductase in resting cells 15th Weurman Flavour Research Symposium, Sep.18-22, Graz, Austria
 Margit Winkler (2016) Aldehyde preparation with new fungal carboxylate reductases 25. Nachwuchswissenschaftlersymposiums Bioorganische Chemie, Sep 21-23, Jena, Germany
 Margit Winkler (2016) New enzymes for the selective enzymatic reduction of carboxylic acids to aldehydes ICC05-AEM2016, Sep 4-8, Toyama, Japan
 Margit Winkler (2016) Carboxylate reductases from fungal Origin for the preparation of aldehyde fragrances, 17th European Congress on Biotechnology, Jul 3-6, Krakow, Poland
 Margit Winkler (2016) New Enzymes for Biocatalytic Syntheses, Symposium Molecular Biotechnology – from the Origins to New Horizons, April 14-15, Graz, Austria
 Margit Winkler (2016) Five Examples of Enzymatic Functional Group Transformations, Mikrobiologisches Seminar, Universität Stuttgart, Feb 3, Stuttgart, Germany.
 Margit Winkler (2016) Formation and transformation of carboxylic acid derivatives, Seminar of the Institute of Applied Synthetic Chemistry, Vienna University of Technology, Jan 14, Vienna, Austria.
 Margit Winkler (2016) Enzymatic Carboxylate Reduction, Institute Seminar of the Institute for Molecular Biotechnology, Graz University of Technology, Jan 13, Graz, Austria
Margit Winkler , Bastian Daniel, Chiam Hashem, Hannah G. Breuer, Ludmila Martinkova, Tea Pavkov-Keller (2023) Carbonyl Reduction by Carboxylic Acid Reductases: An unsurprising Surprise? Biotrans 2023, Jun 25-29, La Rochelle, France
Lukas Schober , Astrid Schiefer, Florian Rudroff, Margit Winkler (2023) Diversification of carotenoid cleaving enzymes serving as ‘ozonylases’ for generation of aldehydes from a lignin-based feedstock. Biotrans 2023, Jun 25- 29, LaRochelle, France
Valentina Jurkaš Hana Dobiašova, Melissa Horvat, Kvetoslava Vranková, Florian Rudroff, Margit Winkler (2022) Flavour of cheese, wine and durian: synthesis of odorous α-hydroxy pentanones with carboligases. European Summit of Industrial Biotechnology 2022, Nov 14- 16, Graz, Austria
Dominic Goj, Stella Ebner, Melissa Horvat, Margit Winkler (2022) Pichia pastoris as a producer of carboxylic acid reductases. European Summit of Industrial Biotechnology 2022, Nov 14- 16, Graz, Austria
Lukas Schober , Florian Rudroff, Margit Winkler (2022) MapADO: Expression and Characterisation of a novel metal-dependent alkene cleaving enzyme. Chemietage 2022, Sep 20- Sep 22, Vienna, Austria
Margit Winkler , Lukas Schober, Dominic Goj, Florian Rudroff (2022) Expanding the toolbox for biocatalytic aldehyde synthesis. Biocat 2022 – 10th International Congress on Biocatalysis, Aug 28- Sep 1, Hamburg, Germany
Melissa Horvat , Astrid Schiefer, Florian Rudroff, Miroslav Pátek, Ludmila Martinková, Margit Winkler (2022) One pot chemoenzymatic route from acid to nitrile. Gordon Research Conference Biocatalysis, Jul 10-15, Manchester, NH, USA
Melissa Horvat , Victoria Weilch, Robert Rädisch, Sebastian Hecko, Astrid Schiefer, Florian Rudroff, Miroslav Pátek, Ludmila Martinková, Margit Winkler (2021) From carboxylic acid to nitrile: a chemoenzymatic route in one pot. Biotrans 2021, Jul 19-21, online
Stella Ebner , Melissa Horvat, Tanja-Saskia Larch, Margit Winkler (2021) E. coli or yeast – which is the better host for microbial carboxylate reduction? Biotrans 2021, Jul 19-21, online
Margit Winkler, Melissa Horvat, Chiam Hashem (2021) With enzymes and microbes to flavor and fragrance molecules, 16th Weurman Flavour Research Symposium, May 3-7, online
Chiam Hashem, Holly Stolterfoht, Claudia Rinnofner, Stefan Steinberger, Margit Winkler, Harald Pichler (2019) Goodbye methanol induction: Secretion of Pseudomonas aeruginosa lipoxygenase by Pichia pastoris upon glycerol feed, European Summit of Industrial Biotechnology, Nov.18-20, Graz, Austria
Maximilian Glänzer , Birgit Grill, Helmut Schwab, Urška Pogorevčnik, Karin Reicher, Kai Donsbach, Daniel Pienaar, Margit Winkler (2019) Enzyme-catalyzed synthesis of amides from nitriles, Österreichische Chemietage, Sep 24-27, Linz, Austria
Florian Rudroff , Margit Winkler, Daniel Schwendenwein, Anna K. Ressmann, Marko D. Mihovilovic, Uwe T. Bornscheuer, Mark Dörr (2019) Random mutagenesis of carboxylate reductase enabled by a substrate independent HT-assay, Biotrans 2019, Jul 7-11, Groningen, The Netherlands
Holly Stolterfoht, David Gradischnig,Margit Winkler (2019) Chimeric carboxylate reductases, Biotrans 2019, Jul 7-11, Groningen, The Netherlands
Birgit Grill, Georg Steinkellner, Gernot Strohmeier, Urška Pogorevčnik, Karin Reicher, Kai Donsbach, Helmut Schwab,Margit Winkler (2019) Towards a new panel of nitrile hydratases, Biotech France, Jun 26-28, Paris, France
Daniel Schwendenwein Anna K. Ressmann, Florian Rudroff, Margit Winkler (2018) A novel assay for the detection of carboxylic acid reductase (CAR) activity in whole cells, 9TH International Congress on Biocatalysis, Aug 26-30, Hamburg, Germany
Florian Rudroff, Margit Winkler Daniel Schwendenwein, Anna K. Ressmann, Marko D. Mihovilovic, Uwe T. Bornscheuer, Mark Dörr (2018) The ABAO assay, a new tool for Carboxylic Acid Reductase engineering, Gordon Research Conference Biocatalysis, Jul 8-12, Biddeford, ME, USA
Florian Rudroff, Margit Winkler Marcello Entner, Daniel Schwendenwein, Anna K. Ressmann, Marko D. Mihovilovic (2018) Fragrance synthesis via a chemo- & multi enzyme cascade, Gordon Research Conference Biocatalysis, Jul 8-12, Biddeford, ME, USA
Daniel Schwendenwein, Florian Rudroff, Margit Winkler (2017) Cascade reactions for aldehyde generation in vivo, European Summit of Industrial Biotechnology, Nov.14-16, Graz, Austria
Florian Rudroff Anna K. Ressmann, Daniel Schwendenwein, Simon Leonhartsberger, Mark Dörr, Marko D. Mihovilovic, Uwe T. Bornscheuer, Margit Winkler, Michael J. Fink, Sofia Milker, Thomas Bayer, Thomas Wiesinger, Nikolin Oberleitner, (2017) Catalytic Cascades – ‘En Route’ to Applied Biochemical Cell Factories, European Summit of Industrial Biotechnology, Nov.14-16, Graz, Austria
Thorsten Bachler, Astrid Weninger, Anton Glieder Margit Winkler, (2017) Cytochrome P450 CYP505 mutants for biooxidations of active pharmaceutical ingredients, World Congress on Oxidation Catalysis, Jul.9-13, Krakow, Poland
Anna K. Ressmann, Simon Leonhartsberger, Holly Stolterfoht, Daniel Schwendenwein, Marko D. Mihovilovic, Margit Winkler, Florian Rudroff (2017) Whole Cell Substrate Screening of Carboxylic Acid Reductases, Biotrans 2017, Jul.9-13, Budapest, Hungary
Daniel Schwendenwein, Giuseppe Fiume, Harald Pichler, Margit Winkler (2017) Carboxylic Acid Reductase (CAR) mediated Biotransformations, Biotrans 2017, Jul.9-13, Budapest, Hungary
Daniel Schwendenwein, Holly Stolterfoht, Florian Rudroff, Margit Winkler (2017) Modulation of NcCAR and PPTase Expression levels for Aldehyde Preparation with whole cells, Biotrans 2017, Jul.9-13, Budapest, Hungary
Holly Stolterfoht, Daniel Schwendenwein, Florian Rudroff, Kinga Rudnicka, Margit Winkler (2017) Identification of Key-Residues for Enzymatic Carboxylate Reduction, BIOTRANS 2017, Jul.9-13, Budapest, Hungary
Thomas Bayer, Sofia Milker, Thomas Wiesinger, Margit Winkler, Marko D. Mihovilovic, Florian Rudroff (2017) Substrate funnelling complements flux optimization tools to maximize product titers of in vivo cascades, BIOTRANS 2017, Jul.9-13, Budapest, Hungary
Daniel Schwendenwein, Phillipe Oberhemm, Giuseppe Fiume, Florian Rudroff, Kerstin Steiner, Kinga Rudnicka, Margit Winkler (2016) Selective preparation of Aldehydes using carboxylate reductases, European Summit of Industrial Biotechnology, Nov.14-16, Graz, Austria
Giuseppe Fiume, Philippe Oberhemm, Daniel Schwendenwein, Margit Winkler (2016) A new fungal carboxylic acid reductase for the preparation of aldehydes from acids, 8th ÖGMBT Annual Meeting, Sep 12-14, Graz, Austria
Daniel Schwendenwein, Florian Rudroff, Margit Winkler (2016) Tuning the expression of Neurospora crassa carboxylic acid reductase towards an efficient whole cell catalyst for aldehyde preparation, 8th ÖGMBT Annual Meeting, Sep 12-14, Graz, Austria.
Christoph K. Winkler, Markus Plank, Margit Winkler, Kurt Faber (2016) Chemo-, regio- and stereoselectivity of carboxylic acid reductases, BIOCAT 2016– 8th International Congress on Biocatalysis, Aug 28-Sep 1, Hamburg, Germany
Daniel Schwendenwein, Florian Rudroff, Kerstin Steiner, Philippe Oberhemm, Giuseppe Fiume, Margit Winkler (2016) Aldehyde preparation with new fungal carboxylate reductases, Gordon Research Conference Biocatalysis, Jul 10-15, Biddeford, ME, USA
Thomas Bayer, Thomas Wiesinger, Sofia Milker, Margit Winkler, Florian Rudroff, Marko D. Mihovilovic (2016) Convergent Carbon Flux via Substrate Funelling for the In Vivo Production of Reactive Aldehyde Species, Gordon Research Conference Biocatalysis, Jul 10-15, Biddeford, ME, USA This Poster was outstanding and selected for a Talk
Margit Winkler, (2015) Enzyme catalyzed reduction of carboxylic acids, BIOTRANS 2015, Jul 26-30, Vienna, Austria This Poster was awarded a ‘Biotrans Poster Award’
Margit Winkler, Martina Geier, Thorsten Bachler, Steven P. Hanlon, Matthias Kittelmann, Anton Glieder, Stephan Lütz, Beat Wirz (2014) Human flavin monooxygenase 2: Heterologous expression in E. coli and API modification, 16th European Congress on Biotechnology, Jul 13-16, Edinburgh, Scotland
Elisa Lanfranchi , Kerstin Steiner, Barbara Darnhofer, Ruth Birner-Grünberger, Anton Glieder, Margit Winkler (2013) A new hydroxynitrile lyase from fern: from the plant to the sequence, Biotrans 2013, Jul 21-25, Manchester, United Kingdom
Kamila Napora, Natalia Pankiewicz, Manoj N. Sonavane, Stefan Krahulec, Patrizia Wildberger, Andrea Camattari, Harald Pichler, Manuela Avi, Margit Winkler (2012) Recombinant expression and characterization of a zinc-dependent alcohol dehydrogenase from Yarrowia lipolytica, BIOCAT 2012– 6th International Congress on Biocatalysis, Sep 2-6, Hamburg, Germany
Margit Winkler, Martin R. Kulterer, Benjamin Stickler, Norbert Klempier (2007) Nitrilase catalyzes key step to γ-amino acids in high optical purity, BIOTRANS 2007, Jul.8-13, Oviedo, Spain
Margit Winkler, Martin Schmuck, Norbert Klempier (2005) Enantioselective microbial transformation of ß-amino nitriles, BIOTRANS 2005, Jul.3-8, Delft, The Netherlands
Birgit Grill, Melissa Horvat, Helmut Schwab, Ralf Gross, Kai Donsbach, Margit Winkler Gordonia hydrophobica Nitrile Hydratase for Amide Preparation from Nitriles, In: Enzymes and Biocatalysis, Editors: Kuo C.H., Huang C.-Y., Shieh C.-J. and Dong C.-D. , 2022, MDPI, pp. 95-104. ISBN: 978-3-0365-5412-9
Margit Winkler, Anna Schwarz, Gernot A. Strohmeier, Jeffrey Kohrt, Roger M. Howard Direct Access to Aldehydes Using Commercially Available Carboxylic Acid Reductases, In: Applied Biocatalysis: The Chemist’s Enzyme Toolkit, Editors: Whittal J., and Sutton P., 2020, Wiley-VCH, pp. 270-276. ISBN: 978-1-119-48701-2
 Margit Winkler Blog article: How to prepare flavors and fragrances by enzymes https://www.acib.at/how-flavors-and-fragrances-are-produced-by-enzymes/, May 18th 2017 
 Martin Walpot Press release: Von der Säure zum Aldehyd, May 12th 2017 

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