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Applied Microbiology and Biotechnology

, Volume 98, Issue 1, pp 83–94 | Cite as

Optimization of arylacetonitrilase production from Alcaligenes sp. MTCC 10675 and its application in mandelic acid synthesis

  • S. K. Bhatia
  • P. K. Mehta
  • R. K. Bhatia
  • T. C. Bhalla
Biotechnological products and process engineering

Abstract

Alcaligenes sp. MTCC 10675 has been isolated from soil sample using enrichment method and has nitrilase catalytic system which is highly specific for the hydrolysis of arylaliphatic nitriles. Optimization of culture conditions using response surface methodology and inducer-mediated approach enhanced arylacetonitrilase production significantly (2.4-fold). Isobutyronitrile acted as an effective inducer for the induction of arylacetonitrilase, and it is highly specific for arylacetonitriles (phenyl acetonitrile and mandelonitrile). Arylacetonitrilase has no effect on its relative velocity (V r) up to 20 mM substrate (mandelonitrile) concentration and at 30 mM mandelonitrile, 23.4 % degree of inhibition (I d) was recorded. Half life of arylacetonitrilase of Alcaligenes sp. MTCC 10675 was 27.5 h at 25 °C. Hg2+, Ag+, Pb3+, and Co2+ were strong inhibitor of arylacetonitrilase activity which resulted into 100 %, 91 %, 84 %, and 83 % inhibition, respectively. Polar protic solvent (dichloromethane, dimethylsulphooxide, and n-butanol) reduce arylacetonitrilase activity up to 80–94 % at 10 % concentration. Alcaligenes sp. MTCC 10675 has higher biocatalytic activity, i.e., 3.9 gg-1 dcw, which is highest in comparison to till reported organism. Arylacetonitrilase-mediated hydrolysis of racemic mandelonitrile resulted into R-(-) mandelic acid with 99.0 % enantiomeric excess (e.e.)

Keywords

Arylacetonitrilase Isobutyronitrile Mandelonitrile Response surface methodology 

Notes

Acknowledgments

The authors acknowledge the Department of Biotechnology and University Grant Commission, India for financial support in the form of Senior Research Fellowship to Mr. Shashi Kant Bhatia, Praveen Kumar Mehta, and Ravi Kant Bhatia. The computational facility availed at Bioinformatics Centre, Himachal Pradesh University is also duly acknowledged.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • S. K. Bhatia
    • 1
  • P. K. Mehta
    • 1
  • R. K. Bhatia
    • 1
  • T. C. Bhalla
    • 1
  1. 1.Department of BiotechnologyHimachal Pradesh UniversityShimlaIndia

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