Expression of nitrile hydratase gene of mutant 4D strain of Rhodococcus rhodochrous PA 34 in Pichia pastoris

Abstract

The nitrile hydratase (NHase) gene of Rhodococcus rhodochrous PA-34 mutant 4D has been amplified by PCR, cloned and expressed in Pichia pastoris KM-71 using pHIL-D2 expression vector. The recombinant P. pastoris KM-71 exhibited active expression of the nitrile hydratase gene of the mutant 4D and has shown very good potential for the transformation of 3-cyanopyridine to nicotinamide. The recombinant P. pastoris KM-71 exhibited maximum NHase activity when cultivated in YPD medium was supplemented with 0.4 mM cobalt ions. The recombinant P. pastoris KM-71 showed maximum nitrile hydratase enzyme production, when incubated at 30 °C for 15 h.

Keywords:

• Mutant
• nitrile hydratase
• recombinant P. pastoris
• Rhodococcus rhodochrous PA-34

Acknowledgements

The authors also wish to acknowledge the internet facilities provided by Sub-Distributed Information Centre, Department of Biotechnology, Himachal Pradesh University, Shimla-5, India.

Declaration of interest

The authors report no declarations of interest. The research work was supported by a research grant from Department of Biotechnology, Government of India, New Delhi, India.