Abstract
The individual and interactive effects of three independent variables i.e. carbon source (glucose), nitrogen source (sodium nitrate) and inducer (ϵ-caprolactam) on nitrilase production from Fusarium proliferatum were investigated using design of experiments (DOE) methodology. Response surface methodology (RSM) was followed to generate the process model and to obtain the optimal conditions for maximum nitrilase production. Based on central composite design (CCD) a quadratic model was found to fit the experimental data (p<0.0001) and maximum activity of 59.0U/g biomass was predicted at glucose concentration (53.22 g/l), sodium nitrate (2.31 g/l) and ϵ-caprolactam (3.58 g/l). Validation experiments were carried out under the optimized conditions for verification of the model. The nitrilase activity of 58.3U/g biomass obtained experimentally correlated to the predicted activity which proves the authenticity of the model. Overall 2.24 fold increase in nitrilase activity was achieved as compared to the activity before optimization (26U/g biomass).
Highlights
Nitriles, organic compounds containing (−CN) functional group, are widespread in the environment
The biocatalytic route is of particular interest because the nitrile-degrading enzymes like nitrilases, amidases and nitrile hydratases (−NHases) can convert diverse nitrile substrates into various amides and carboxylic acids under relatively
It was found that our strain Fusarium proliferatum AUF-2 produces maximum nitrilase (26U/g biomass) with glucose as carbon source, sodium nitrate as nitrogen source and ε-caprolactam as inducer
Summary
Organic compounds containing (−CN) functional group, are widespread in the environment. Occurring nitriles mainly comprise cyanoglycosides, cyanolipids and phenylacetonitrile (Marron et al 2012) that are produced as defensive metabolites in plants. Nitriles are extensively used as feedstocks, solvents, polymers, pharmaceuticals, pesticides and drug intermediates. Nitriles are important intermediates in the organic synthesis of amides, amines, carboxylic acids, esters and ketones (Gong et al 2012; Kobayashi and Shimizu 2000). Bio- and chemo-catalyzed transformations of nitrile substrates lead to the formation of several industrially important amides and organic acids, such as acrylamide, nicotinamide, acrylic and mandelic acids, methacrylic acid (Xue et al 2011). The biocatalytic route is of particular interest because the nitrile-degrading enzymes like nitrilases, amidases and nitrile hydratases (−NHases) can convert diverse nitrile substrates into various amides and carboxylic acids under relatively
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