Abstract
Objectives
To identify a novel nitrilase with S-selectivity toward mandelonitrile that can produce (S)-mandelic acid in one step.
Results
A novel nitrilase PpL19 from Pseudomonas psychrotolerans L19 was discovered by genome mining. It showed S-selectivity with an enantiomeric excess of 52.7 % when used to hydrolyse (R, S)-mandelonitrile. No byproduct was observed. PpL19 was overexpressed in Escherichia coli BL21 (DE3) and formed inclusion bodies that were active toward mandelonitrile and stable across a broad range of temperature and pH. In addition, PpL19 hydrolysed nitriles with diverse structures; arylacetonitriles were the optimal substrates. Homology modelling and docking studies of both enantiomers of mandelonitrile in the active site of nitrilase PpL19 shed light on the enantioselectivity.
Conclusions
A novel nitrilase PpL19 from P. psychrotolerans L19 was mined and distinguished from other nitrilases as it was expressed as an active inclusion body and showed S-selectivity toward mandelonitrile.
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Acknowledgments
This work was supported by the National Natural Science Foundation of China (No. 21406068/B060804), China Postdoctoral Science Foundation funded project (No. 2014M560308) and National Major Science and Technology Projects of China (2012ZX09304009).
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Sun, H., Gao, W., Wang, H. et al. Expression, characterization of a novel nitrilase PpL19 from Pseudomonas psychrotolerans with S-selectivity toward mandelonitrile present in active inclusion bodies. Biotechnol Lett 38, 455–461 (2016). https://doi.org/10.1007/s10529-015-1992-0
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DOI: https://doi.org/10.1007/s10529-015-1992-0