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 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 and and a new Fe-type NHase from were very well expressed and hydrated methacrylonitrile, pyrazine-carbonitrile, and 3-amino-3-(-toluoyl)propanenitrile. The Co-dependent NHases from and , as well as two Fe-dependent NHases from , were-in addition-able to produce the amide from cinnamonitrile. Summarizing, seven so far uncharacterized NHases are described to be promising biocatalysts.
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| http://dx.doi.org/10.3390/molecules25112521 | DOI Listing |
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