Characterization of nitrilase-producing thermophilic bacteria and exploring their potential for mandelic acid synthesis

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Research Articles | Published:

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Pub Email:
Doi: 10.1007/s42535-023-00603-7
First Page: 257
Last Page: 265
Views: 1141

Keywords: Thermophile, Nitrilase, n Geobacillusn , Mandelic acid, Enantioselectivity


The present study was undertaken to screen thermophilic nitrilase-producing bacteria from the hot water springs of Manikaran and Kheerganga, located in the Kullu District of Himachal Pradesh, India, to harness mandelic acid from mandelonitrile. Five nitrile degrading isolates from these thermophilic sites exhibiting appreciable mandelonitrile degrading ability were subjected to biochemical characterization, and all bacteria were gram-positive rods. Molecular characterization was done by 16s rDNA sequencing, and the phylogenetic tree depicted their relatedness. There were three bacteria: Bacillus licheniformis KMA 1, Lysinibacillus macroides KMA 2, and Bacillus pumilus KMA 3 from Kheerganga. On the other hand, two promising bacteria were obtained from the Manikaran hotspring, designated as Geobacillus stearothermophilus MAC I and Geobacillus icigianus MAC VI, displayed high thermotolerance and suitably grew at 60 °C. Geobacillus strains showed a similar enzymatic profile when tested against mandelonitrile as substrate. Therefore, only one strain, G. icigianus MAC VI, was selected for optimization and scale-up studies. Even at elevated temperatures of 100 °C, the enzyme retained its enzymatic prowess and could effectively degrade mandelonitrile; however, a dip in enzyme activity was also concurrently recorded. The standardization of process parameters further resulted in a 7.4-fold increase in the nitrilase activity of the selected strain. Furthermore, a delayed inducer feeding strategy was also incorporated to cause hyper-induction of nitrilase enzyme effectively. Scale-up production of mandelic acid was carried out in a batch reaction that yielded 8.4 g of R-mandelic acid with 74%ee.

Thermophile, Nitrilase, n              Geobacillusn            , Mandelic acid, Enantioselectivity

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Authors are highly thankful to the Honorable Vice-Chancellor, Prof. P.K. Khosla, for giving them the required facilities to conduct the research. This work was supported by the Department of Science and Technology, Government of India (grant number SB/YS/LS-19/2013).

Author Information

Sharma Mamta
School of Biotechnology, Sher-e-Kashmir University of Agricultural Sciences & Technology of Jammu, Chatha, Jammu, India

Attri Chandrika
Faculty of Applied Sciences and Biotechnology, Shoolini University, Solan, India

Seth Amit
Department of Life Sciences (Botany), Manipur University, Imphal, Manipur, India