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Recent advances and challenges in the heterologous production of microbial nitrilases for biocatalytic applications


The aim of this study is to review the current state of and highlight the challenges in the production of microbial nitrilases as catalysts for the mild hydrolysis of industrially important nitriles. Together with aldoxime dehydratase, the nitrile-hydrolyzing enzymes (nitrilase, nitrile hydratase) are key enzymes in the aldoxime–nitrile pathway which is widely distributed in bacteria and fungi. The availability of nitrilases has grown significantly over the past decade due to the use of metagenomic and database-mining approaches. Databases contain plenty of putative enzymes of this type, whose overproduction may improve the spectrum and the industrial utility of nitrilases. By exploiting this resource, the number of experimentally verified nitrilases has recently increased to several hundred. We especially focus on the efficient heterologous expression systems that are applicable for the overproduction of wild-type nitrilases and their artificial variants. Biocatalyst forms with industrial potential are also highlighted. The potential industrial applications of nitrilases are classified according to their target products (α-hydroxy acids, α- and β-amino acids, cyano acids, amides). The emerging uses of nitrilases and their subtypes (cyanide hydratases, cyanide dihydratases) in bioremediation is also summarized. The integration of nitrilases with other enzymes into artificial multienzymatic and chemoenzymatic pathways is considered a promising strategy for future applications.

Graphical Abstract

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Fig. 1
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Cyanide hydratase


Cross-linked enzyme aggregates


Cyanide dihydratase


Dry cell weight


Enantiomeric excess




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This study was funded by the Ministry of Education, Youth and Sports of the Czech Republic (project COST LD15107) and the Institute of Microbiology of the Academy of Sciences of the Czech Republic, v.v.i. (project RVO61388971).

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Correspondence to Ludmila Martínková.

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Martínková, L., Rucká, L., Nešvera, J. et al. Recent advances and challenges in the heterologous production of microbial nitrilases for biocatalytic applications. World J Microbiol Biotechnol 33, 8 (2017).

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  • Aldoxime–nitrile pathway
  • Biocatalytic applications
  • Database mining
  • Heterologous production
  • Metagenome mining
  • Nitrilase