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A novel nitrilase from Ralstonia eutropha H16 and its application to nicotinic acid production

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Abstract

A novel aliphatic nitrilase, REH16, was found in Ralstonia eutropha H16 and overexpressed in Escherichia coli BL21(DE3), and its enzymatic properties were studied. The temperature and pH optima were 37 °C and 6.6, respectively, and the best thermostability of the nitrilase was observed at 25 °C, which preserved 95% of activity after 120 h of incubation. REH16 has a broad hydrolytic activity toward aliphatic and heterocyclic nitriles and showed high tolerance of 3-cyanopyridine; this enzyme could hydrolyze as high as 100 mM 3-cyanopyridine completely. To improve the 3-cyanopyridine conversion efficiency in an aqueous reaction system, water-miscible organic solvents were tested, and ethanol (10% v/v) was chosen as the optimal co-solvent. Finally, under optimized conditions, using the fed-batch reaction mode, total of 1050 mM 3-cyanopyridine was hydrolyzed completely in 20.8 h with eight substrate feedings, yielding 129.2 g/L production of nicotinic acid and thus showing a potential for industrial application.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant No. 21406068/B060804), the Fundamental Research Funds for the Central Universities.

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Authors and Affiliations

  1. State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, People’s Republic of China

    Haiyang Fan, Lifeng Chen, Hualei Wang, Yuhong Ren & Dongzhi Wei

  2. Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, 266071, Shandong, People’s Republic of China

    Huihui Sun

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  1. Haiyang Fan
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  2. Lifeng Chen
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  4. Hualei Wang
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Correspondence to Hualei Wang.

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Fan, H., Chen, L., Sun, H. et al. A novel nitrilase from Ralstonia eutropha H16 and its application to nicotinic acid production. Bioprocess Biosyst Eng 40, 1271–1281 (2017). https://doi.org/10.1007/s00449-017-1787-x

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