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Cross-linked enzyme aggregates of arylamidase from Cupriavidus oxalaticus ICTDB921: process optimization, characterization, and application for mitigation of acrylamide in industrial wastewater

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Abstract

Acrylamidase produced by Cupriavidus oxalaticus ICTDB921 was recovered directly from the fermentation broth by ammonium sulfate (40–50%) precipitation and then stabilized by cross-linking with glutaraldehyde. The optimum conditions for the preparation of cross-linked enzyme aggregates of acrylamidase (acrylamidase-CLEAs) were using 60 mM glutaraldehyde for 10 min at 35 °C and initial broth pH of 7.0. Acrylamidase-CLEAs were characterized by SDS-PAGE, FTIR, particle size analyzer and SEM. Cross-linking shifted the optimal temperature and pH from 70 to 50 °C and 5–7 to 6–8, respectively. It also altered the secondary structure fractions, pH and thermal stability along with the kinetic constants, Km and Vmax, respectively. A complete degradation of acrylamide ~ 1.75 g/L in industrial wastewater was achieved after 60 min in a batch process under optimum operating conditions, and the kinetics was best represented by Edward model (R2 = 0.70). Acrylamidase-CLEAs retained ~ 40% of its initial activity after three cycles for both pure acrylamide and industrial wastewater, and were stable for 15 days at 4 °C, retaining ~ 25% of its original activity.

Article highlights

  • Acrylamidase from broth was recovered as cross linked enzyme aggregates (CLEAs)

  • Acrylamidase-CLEAs retained good kinetic and pH/thermal stability

  • Acrylamidase-CLEAs possessed good storage stability and reusability

  • Acrylamidase-CLEAs efficiently degraded acrylamide from industrial waste water

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Acknowledgements

The authors would like to thank the Department of Biotechnology, Ministry of Science and Technology, Government of India, New Delhi, for providing the financial supports to carry out this research work.

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  1. Food Engineering and Technology Department, Institute of Chemical Technology, Matunga, Mumbai, 400019, India

    Nidhi H. Kulkarni, Abhijeet B. Muley, Dattatray K. Bedade & Rekha S. Singhal

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Kulkarni, N.H., Muley, A.B., Bedade, D.K. et al. Cross-linked enzyme aggregates of arylamidase from Cupriavidus oxalaticus ICTDB921: process optimization, characterization, and application for mitigation of acrylamide in industrial wastewater. Bioprocess Biosyst Eng 43, 457–471 (2020). https://doi.org/10.1007/s00449-019-02240-4

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