COST Action WG D25/0002/02

  • Start of Action – 26/10/2001; End of Action – 25/04/2007
  • Theme Abstract

Nitrile- and amide-hydrolyzing enzymes as tools in organic chemistry

Nitriles and amides are widely exploited as building blocks in organic synthesis. The problems associated with their chemical hydrolysis (low yields, poor regio-, chemo- and stereoselectivity) can be overcome by the use of enzymes – nitrile hydratases, nitrilases and amidases. This project aims to introduce nitrile-converting microorganisms with new substrate specificities and improved regio-, chemo- and stereoselecti­vities. Physico-chemical modifications (immobilization, lyophilization, solubilization and treatment by chemical reagents of the enzymes) and medium-engineering (particularly application of organic cosolvents) will be utilized in order to enhance the potential of nitrile and amide biotransformations. The introduction of novel biocatalysts and processes is expected to enable major improvements in the production of fine and pharmaceutical chemicals. Besides, the project will provide background knowledge applicable in the reduction of environmental pollution caused by toxic nitrile contaminants.


  • Final Report

  1. New microbial strains and enzymes were discovered and largely made available to WG partners. These new biocatalysts comprise the whole spectrum of enzymes involved in hydrolytic nitrile degradation pathways (nitrile hydratase, amidase, nitrilase, aldoxime dehydratase; see Scheme 5) and secondary amide hydrolase.
  2. Different novel approaches proved to be viable for immobilization of nitrile- and amideconverting enzymes (retention of nitrilase in an ultrafiltration membrane bioreactor, crosslinked enzyme aggregate technology, adsorption to hydrophobic supports like Butyl Sepharose) and to their stabilization (via co-aggregation with polyethyleneimine). The first two methods have been applied during the STSMs (O. Kaplan to Univ. L´Aquila, D. Kubá_ to TU Delft).
  3. A broad spectrum of new substrates including chiral compounds (e.g., substituted cyclopentane- and cyclohexanecar­bonitriles, 3-hydroxynitriles, 2-oxonitriles, 2-halonitriles and 2-amino nitriles; hexosamine derivatives) bearing different protecting groups have been prepared as starting materials for the synthesis of useful synthons and their transformation accomplished in a chemo-, regio- or stereoselective way using the above enzymes.
  4. The above achievements have been largely published (17 articles, 2 book chapters publishhed or accepted; further three joint articles are in preparation – to be submitted by the end of 2007). In addition, WG2 presented about 25 joint conference communications. Please see below for a list of joint articles, book chapters and abstracts.
  5. The activities of the WG promoted the participation of young microbiologists, biochemists and organic chemists. Throughout WG2, the staffs contained a high ratio of postgraduate students and postdocs. Undergraduate students also participated in the research. The young scientists significantly contributed to the publication output of WG2. The participation in the D25 Action enabled them to present their works at workshops and WG meetings. Particularly, the short-term scientific missions were important for the enhancement of qualification of the young scientists. All the six STSMs were accomplished by PhD students and afforded a valuable output, as documented by the STSM reports. These efforts were also supported by a bilateral project between WG members: Bilateral collaboration between the Lab. of Biotransformation Prague and University Hohenheim has been granted with financial support from DAAD and the Acad. Sci. Czech Rep. for 2006–2007. The topic of this project (“Preparation of chiral synthons from nitriles”) is a continuation of the WG2 programme and enabled other four student exchanges in 2006–2007 (in total duration of about 12 weeks).


  • Kick-off meeting: October 19, 2002, University of Stuttgart

    Final Programme

    9.00 – 9.10 Welcome address (Christoph Syldatk)
    Morning session – Chair: Christoph Syldatk
    09.10 – 09.20 Introductory remarks – short-term scientific missions, workshops (Ludmila Martínková)
    09.20 – 09.40 Georg Gübitz (Graz University of Technology, Graz, Austria
    09.40 – 10.00 Artur Cavaco-Paulo (University of Minho, Guimarães, Portugal)
    10.00 – 10.20 Pedro F. B. Brandao (University of Stuttgart, Stuttgart, Germany) „Genetic diversity of nitrile hydratase and amidase enzymes in geographically distinct Rhodococcus erythropolis strains „
    10.20 – 10.50 Coffee Break
    Chair: Ludmila Martínková
    10.50 – 11.10 Lutz Fischer (University of Stuttgart-Hohenheim, Stuttgart, Germany)
    11.10 – 11.30 Fred van Rantwijk (Delft University of Technology, Delft, NL
    11.30 – 11.50 Pedro Lourenço (INETI, Lisbon, Portugal)
    11.50 – 13.00 Lunch Break
    Afternoon session – Chair: Lutz Fischer
    13.00 – 13.20 Maria Cantarella (University of L´Aquila, L´Aquila, Italy)
    13.20 – 13.40 Giovanni Nicolosi (Institute of Natural Substances CNR, Valverde, Italy)
    13.40 – 14.00 Ludmila Martínková (Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic)
    14.00 – 14.30 Coffee Break
    Chair: Fred van Rantwijk
    14.30 – 14.50 Jean Bolte (University of Blaise Pascal, SEESIB, Aubiere, France)
    14.50 – 15.10 Karel Bezouška (Charlers University, Prague, Czech Republic)
    15.10 – 15.30 Closing Remarks
    16.00 – 17.30 Excursion to the University of Hohenheim, Stuttgart (Laboratories, pilot plant facilities and botanical garden)


  • WG meeting: July 3, 2005, TU Delft (in conjunction with Biotrans 2005)

Delft University of Technology,
Laboratory of Biocatalysis and Organic Chemistry
Julianalaan 136,
2628 BL Delft,
The Netherlands

3rd July 2005

Metting programme

10.30–10.40 – Fred van Rantwijk (TU Delft, The Netherlands)
Morning session – Chair: Vicente Gotor (Univ. Oviedo, Spain)
10.40–11.00 – Ludmila Martinkova (Institute of Microbiology, Prague, Czech Rep.)
11.00–11.30 – 11.00–11.30 Andrzej Chmura, Bruno Fernandes, Ceasar Mateo, Fred van Rantwijk, Roger Sheldon (TU Delft, The Netherlands)
11.30–12.00 – Norbert Klempier, Margit Winkler, Astrid Knall (TU Graz, Austria)
12.00–12.30 – Maria Cantarella, Laura Cantarella, Agata Spera (Univ. L´Aquila, Italy)
12.30–13.30 Lunch
Afternoon session – Chair: Lutz Fischer (Univ. Hohenheim, Germany)
13.30–14.00 – Miguel Ferrero, Susana Fernández, Vicente Gotor (Univ. Oviedo, Spain)
14.00–14.30 – Ondřej Kaplan, Vojtech Vejvoda, Maria Cantarella, Ludmila Martinkova (Institute of Microbiology, Prague, Czech Rep.)
14.30–15.00 Discussions

Meeting participants photo


  • WG meeting: April 18, 2006, Graz, Austria

Meeting venue

Graz University of Technology,
Applied Biocatalysis Building (1st floor),
Petersgasse 14, A-8010 Graz, Austria

Metting programme

9.30–9.40 – Norbert Klempier (TU Graz, Austria)
9.40–9.45 – Ludmila Martinkova (Institute of Microbiology, Prague, Czech Rep.)
Session I – Chair: Ludmila Martínková
9.45–10.10 – Sander van Pelt, Andrzej Chmura, Fred van Rantwijk (TU Delft, The Netherlands)
10.10–10.35 – Norbert Klempier, Margit Winkler (TU Graz-Inst. Org. Chem., Austria)
10.35–11.00 – Maria Cantarella, Alberto Gallifuoco, Agatha Spera, Laura Cantarella, Ondrej Kaplan and Ludmila Martínková (Univ. L´Aquila, Italy, Univ. Cassino, Italy, Institute of Microbiology, Prague, Czech Rep.)
11.00–11.30 Coffee break
Session II – Chair: Sabine Lutz-Wahl
11.30–11.55 – Georg Gübitz (TU Graz, Austria)
11.55–12.20 – Carla Silva, Artur Cavaco-Paulo (Univ. Minho, Portugal)
11.20–12.45 – Vicente Gotor-Fernández, Eduardo Busto and Vicente Gotor (Univ. Oviedo)
12.45–14.30 Lunch
Session III – Chair: Fred van Rantwijk
14.30–14.55 – El Blidi Lahssen, Bolte Jean and Lemaire Marielle (B. Pascal Univ., Aubière, France)
14.55–15.20 – David Kubáč, Ondřej Kaplan, Vojtěch Vejvoda, Maria Cantarella, Laura Cantarella, Marielle Lemaire, Jean Bolte and Ludmila Martinkova (Institute of Microbiology, Prague, Czech Rep.; Univ. L´Aquila, Italy; Univ. Cassino, Italy; B. Pascal Univ., Aubière, France)
15.20–15.45 Discussion

Poster session (in conjunction with the MECP Symposium Graz ´06)

  • O.Kaplan, V. Vejvoda, M. Cantarella, L. Cantarella, V. Křen, L. Martínková (Institute of Microbiology, Prague, Czech Rep.; Univ. L´Aquila, Italy; Univ. Cassino, Italy):
  • M. Cantarella, L. Cantarella, A. Gallifuoco, A. Spera, O.Kaplan, L. Martínková (Univ. L´Aquila, Italy; Univ. Cassino, Italy; Institute of Microbiology, Prague, Czech Rep.):
  • D.Kubáč, O. Kaplan, M. Himl, L. Martínková (Institute of Microbiology Prague, Czech Rep.):
  • M. Wälz, M.L. Magri, S. Lutz-Wahl, L. Fischer (University of Hohenheim, Germany):
  • E. Sagot, B. Aboab, J. Bolte, T. Gefflaut (B. Pascal Univ., Aubière, France):