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A Comparative Study of Nitrilases Identified by Genome Mining

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Abstract

Escherichia coli strains expressing different nitrilases transformed nitriles or KCN. Six nitrilases (from Aspergillus niger (2), A. oryzae, Neurospora crassa, Arthroderma benhamiae, and Nectria haematococca) were arylacetonitrilases, two enzymes (from A. niger and Penicillium chrysogenum) were cyanide hydratases and the others (from P. chrysogenum, P. marneffei, Gibberella moniliformis, Meyerozyma guilliermondi, Rhodococcus rhodochrous, and R. ruber) preferred (hetero)aromatic nitriles as substrates. Promising nitrilases for the transformation of industrially important substrates were found: the nitrilase from R. ruber for 3-cyanopyridine, 4-cyanopyridine and bromoxynil, the nitrilases from N. crassa and A. niger for (R,S)-mandelonitrile, and the cyanide hydratase from A. niger for KCN and 2-cyanopyridine.

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Abbreviations

aa:

Amino acid

NitAb:

Nitrilase from Arthroderma benhamiae CBS 112371 (GenBank: EFE30690)

NitAn:

Nitrilase from Aspergillus niger CBS 513.88 (GenBank: CAK46742)

NitAn1:

Nitrilase from Aspergillus niger K10 (CCF 3411 GenBank: ABX75546)

NitAn2:

Nitrilase from Aspergillus niger CBS 513.88 (GenBank: CAK47246)

NitAo:

Nitrilase from Aspergillus oryzae RIB40 (GenBank: BAE63579)

NitGm:

Nitrilase from Giberella moniliformis (GenBank: ABF83489)

NitMg:

Nitrilase from Meyerozyma guilliermondi (Pichia guilliermondi) ATCC 6260 (NCBI Reference Sequence: XP_001482890)

NitNc:

Nitrilase from Neurospora crassa OR74A (GenBank: CAD70472)

NitNh:

Nitrilase from Nectria haematococca mpVI 77-13-4 (GenBank: EEU45207)

NitPc1:

Nitrilase from Penicillium chrysogenum Wisconsin 54-1255 (NCBI Reference Sequence: XP_002562104)

NitPc2:

Nitrilase from Penicillium chrysogenum Wisconsin 54-1255 (NCBI Reference Sequence: XP_002565836)

NitPm:

Nitrilase from Penicillium marneffei ATCC18224 (NCBI Reference Sequence: XP_002144951)

NitRr1:

T365S/L366I variant of nitrilase from Rhodococcus ruber PA-34 (EMBL Bank: HF543938)

NitRr2:

T365S/L366I variant of nitrilase from Rhodococcus ruber NHB-2 (EMBL Bank: HF543939)

NitRr3:

T365S/L366I variant of nitrilase from Rhodococcus rhodochrous NDB 1165 (EMBL Bank: HF543940)

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Acknowledgments

This study was supported via projects P504/11/0394 (Czech Science Foundation), TA01021368 (Technology Agency of the Czech Republic) and internal project RVO61388971 (Institute of Microbiology).

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Author notes

  1. Ondřej Kaplan

    Present address: Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Heyrovského nám 2, 162 06, Prague, Czech Republic

Authors and Affiliations

  1. Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220, Prague, Czech Republic

    Ondřej Kaplan, Alicja B. Veselá, Alena Petříčková, Fabrizia Pasquarelli, Martina Pičmanová, Anna Rinágelová, Miroslav Pátek & Ludmila Martínková

  2. Department of Biochemistry, Faculty of Science, Charles University in Prague, Hlavova 8, 12840, Prague, Czech Republic

    Alicja B. Veselá & Alena Petříčková

  3. Department of Chemical Engineering, University of L’Aquila, via Campo di Pile, 67100, L’Aquila, Italy

    Fabrizia Pasquarelli

  4. Department of Biotechnology, Himachal Pradesh University, Summer Hill, Shimla, 171005, Himachal Pradesh, India

    Tek Chand Bhalla

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  1. Ondřej Kaplan
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Correspondence to Ludmila Martínková.

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12033_2013_9656_MOESM1_ESM.doc

Supplemental file 1 Nucleotide sequences of the nitrilase genes adapted for expression in Escherichia coli using the Generay (Shanghai, China) software (DOC 30 kb)

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Kaplan, O., Veselá, A.B., Petříčková, A. et al. A Comparative Study of Nitrilases Identified by Genome Mining. Mol Biotechnol 54, 996–1003 (2013). https://doi.org/10.1007/s12033-013-9656-6

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