Evaluation of the efficiency of thermostable L-asparaginase from B. licheniformis UDS-5 for acrylamide mitigation during preparation of French fries.

A thermostable L-asparaginase was produced from Bacillus licheniformis UDS-5 (GenBank accession number, OP117154). The production conditions were optimized by the Plackett Burman method, followed by the Box Behnken method, where the enzyme production was enhanced up to fourfold. It secreted L-asparaginase optimally in the medium, pH 7, containing 0.5% (w/v) peptone, 1% (w/v) sodium chloride, 0.15% (w/v) beef extract, 0.15% (w/v) yeast extract, 3% (w/v) L-asparagine at 50 °C for 96 h. The enzyme, with a molecular weight of 85 kDa, was purified by ion exchange chromatography and size exclusion chromatography with better purification fold and percent yield. It displayed optimal catalysis at 70 °C in 20 mM Tris-Cl buffer, pH 8. The purified enzyme also exhibited significant salt tolerance too, making it a suitable candidate for the food application. The L-asparaginase was employed at different doses to evaluate its ability to mitigate acrylamide, while preparing French fries without any prior treatment. The salient attributes of B. licheniformis UDS-5 L-asparaginase, such as greater thermal stability, salt stability and acrylamide reduction in starchy foods, highlights its possible application in the food industry.

Immobilization of α-amylase on GO-magnetite nanoparticles for the production of high maltose containing syrup.

Robust amylases with stability and catalysis at multitude of extremities are the need of an hour. Enzyme immobilization may prove beneficial at commercial scale to achieve such attributes. In the present study, a commercially available amylase was immobilized on graphene oxide (GO) - magnetite (Fe3O4) nanoparticles through covalent bonding. The structural and morphological characterizations were conducted by XRD, SEM and TEM. Further, FTIR and TGA confirmed the interaction between amylase, GO and nanoparticles. The variables, such as concentrations of GO (1.3 mg), Fe3O4 (58 µg), and amylase (4.5 mg) were optimized by the response surface methodology using central composite design. High loading capacity of 77.58 µg amylase over 1 µg GO-magnetite nanoparticles was achieved under optimum conditions. Biochemically, the pH optimum remained unaltered, i.e., pH 7, whereas, the alkalitolerance was increased by ~20% in relative activities upon immobilization. The half-life of soluble amylase was 13 h, which enhanced to 20 h upon immobilization in 20 mM phosphate buffer, pH 7 at 50 °C. Besides, the thermodynamic parameters supported the stability trends. The immobilized amylase could be used for 11 subsequent cycles. The mentioned attributes and the dextrose equivalent values during the production of high maltose containing syrup highlighted its commercialization.