Genipin crosslinked L-asparaginase CLEAs effectively mitigate acrylamide in deep-fried sweet potato chips

Abstract

Crosslinked enzyme aggregates of L-asparaginase (A-CLEAs) were prepared using genipin, a non-toxic crosslinker. The process parameters for the preparation of A-CLEAs were optimized using a 3-level factorial design, which predicted the optimized conditions to be 60 mg/mL BSA, 2 mg/mL genipin and a crosslinking time of 4 h. The model which was validated experimentally, resulted in a recovery of 90% activity of L-asparaginase. While the optimum pH shifted to a slightly acidic range, the optimum temperature remained unchanged. A-CLEAs displayed the best pH stability at pH 9 (48%) after 24 h of incubation. At 50 °C, the deactivation rate constant (Kd) of A-CLEA decreased by 24%. A-CLEAs showed excellent reusability, retaining 79% of their initial activity after 10 cycles of use. The formation of a dark-blue coloured pigmented A-CLEA and the presence of the carboxymethyl group of genipin in FTIR spectrum confirmed the crosslinking reaction. The increase in the β-sheet content of A-CLEA, correlated with its improved stability. The improved thermostability with regards to structural integrity was well established by TGA-DSC analyses. Morphologically, A-CLEAs showed a highly clustered ball-like appearance. As a model example, the ability of genipin crosslinked A-CLEAs to reduce acrylamide in sweet potato chips was checked. The asparagine content of sweet potato was found to be 2906 ± 15.02 mg/kg as quantified by HPTLC. Analysis of acrylamide by GC-FID showed a 94% reduction in sweet potato chips on contact of sweet potato slices with A-CLEAs for 40 min.