Functional Expression and Characterization of a Panel of Cobalt and Iron-Dependent Nitrile Hydratases

Birgit Grill, Maximilian Glänzer, Helmut Schwab, Steiner Kerstin, Daniel Pienaar, Dean Brady, Kai Donsbach, Margit Winkler*

*Korrespondierende/r Autor/in für diese Arbeit

Publikation: Beitrag in einer FachzeitschriftArtikel


Nitrile hydratases (NHase) catalyze the hydration of nitriles to the corresponding amides. We report on the heterologous expression of various nitrile hydratases. Some of these enzymes have been investigated by others and us before, but sixteen target proteins represent novel sequences. Of 21 target sequences, 4 iron and 16 cobalt containing proteins were functionally expressed from Escherichia coli BL21 (DE3) Gold. Cell free extracts were used for activity profiling and basic characterization of the NHases using the typical NHase substrate methacrylonitrile. Co-type NHases are more tolerant to high pH than Fe-type NHases. A screening for activity on three structurally diverse nitriles was carried out. Two novel Co-dependent NHases from Afipia broomeae and Roseobacter sp. and a new Fe-type NHase from Gordonia hydrophobica were very well expressed and hydrated methacrylonitrile, pyrazine-carbonitrile, and 3-amino-3-(p-toluoyl)propanenitrile. The Co-dependent NHases from Caballeronia jiangsuensis and Microvirga lotononidis, as well as two Fe-dependent NHases from Pseudomonades, were—in addition—able to produce the amide from cinnamonitrile. Summarizing, seven so far uncharacterized NHases are described to be promising biocatalysts.
PublikationsstatusVeröffentlicht - 28 Mai 2020

ASJC Scopus subject areas

  • !!Drug Discovery
  • Analytische Chemie
  • !!Chemistry (miscellaneous)
  • !!Molecular Medicine
  • !!Physical and Theoretical Chemistry
  • !!Pharmaceutical Science
  • Organische Chemie

Fields of Expertise

  • Human- & Biotechnology

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