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Biotransformation of 3-cyanopyridine to nicotinic acid using whole-cell nitrilase of Gordonia terrae mutant MN12

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Abstract

In the present study, the Gordonia terrae was subjected to chemical mutagenesis using ethyl methane sulfonate (EMS) and methyl methane sulfonate (MMS), N-methyl-N-nitro-N-nitrosoguanidine (MNNG), 5-bromouracil (5-BU) and hydroxylamine with the aim of improving the catalytic efficiency of its nitrilase for conversion of 3-cyanopyridine to nicotinic acid. A mutant MN12 generated with MNNG exhibited increase in nitrilase activity from 0.5 U/mg dcw (dry cell weight) (in the wild G. terrae) to 1.33 U/mg dcw. Further optimizations of culture conditions using response surface methodology enhanced the enzyme production to 1.2-fold. Whole-cell catalysis was adopted for bench-scale synthesis of nicotinic acid, and 100% conversion of 100 mM 3-cyanopyridine was achieved in potassium phosphate buffer (0.1 M, pH 8.0) at 40 °C in 15 min. The whole-cell nitrilase of the mutant MN12 exhibited higher rate of product formation and volumetric productivity, i.e., 24.56 g/h/g dcw and 221 g/L as compared to 8.95 g/h/g dcw and 196.8 g/L of the wild G. terrae. The recovered product was confirmed by HPLC, FTIR and NMR analysis with high purity (> 99.9%). These results indicated that the mutant MN12 of G. terrae as whole-cell nitrilase is a very promising biocatalyst for the large-scale synthesis of nicotinic acid.

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All data generated or analysed during this study are included in this published article.

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Acknowledgements

The authors acknowledge the Indian Council of Medical Research (ICMR), New Delhi, India (F. No. 3/1/3/JRF-2017/HRD-LS/50606/02), and the University Grants Commission (UGC), New Delhi, India (F. No. 18-1/2011 (BSR)/24th Feb 2014), for financial support in the form of Senior Research Fellowship (SRF) to Monika and Basic Scientific Research-Faculty Fellowship (BSR-FF) to Tek Chand Bhalla, respectively.

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  1. Department of Biotechnology, Himachal Pradesh University, Shimla, Himachal Pradesh, 171005, India

    Monika,  Sheetal, Neerja Thakur & Tek Chand Bhalla

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  1. Monika
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  2. Sheetal
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  3. Neerja Thakur
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  4. Tek Chand Bhalla
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Contributions

The first draft of the manuscript was written by Monika. Design of experiment, analysis and interpretation of data were performed by Monika, NT, Sheetal and TCB. TCB corrected and approved the final manuscript.

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Correspondence to Tek Chand Bhalla.

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Monika, Sheetal, Thakur, N. et al. Biotransformation of 3-cyanopyridine to nicotinic acid using whole-cell nitrilase of Gordonia terrae mutant MN12. Bioprocess Biosyst Eng 46, 195–206 (2023). https://doi.org/10.1007/s00449-022-02823-8

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