1. Cross-linked arginine deiminase aggregates for enhanced production of nutraceutical citrulline.
- Author
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Kawatra, Anubhuti, Chhabra, Deepak, and Gulati, Pooja
- Subjects
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ARGININE deiminase , *CITRULLINE , *INDUSTRIAL efficiency , *GENETIC algorithms , *INDUSTRIAL costs , *GLUTARALDEHYDE - Abstract
Arginine deiminase (ADI) has been widely employed for the biological production of nutraceutical citrulline. However, sub-optimal enzyme properties, operational instability, and difficulty in reuse impede its industrial usage as well as increase the production cost. The present study describes a simple, cost-effective carrier-free immobilization technology that can circumvent these limitations. ADI was precipitated and cross-linked with glutaraldehyde to obtain novel cross-linked arginine deiminase aggregates (ADI-CLEA). A neural network-based multiobjective genetic algorithm (MOGA-NN) was employed to optimize the immobilization of ADI. MOGA-NN optimization yielded ADI-CLEA with 0.30 IU/mL activity and 61 % recovery. Structural characterization using TEM, SEM, FTIR, DLS/ZETA revealed that the developed ADI-CLEA were monodisperse, ultra-porous, and highly stable. ADI-CLEA showed an improved pH /temperature profile and resistivity against denaturants than free enzyme. The efficiency of ADI-CLEA in the industrial production of nutraceutical citrulline was also studied. ADI-CLEA produced 215.4 g/L citrulline in first cycle of usage. Reusability and storage stability analysis further showed that ADI-CLEA could be reused for more than seven consecutive cycles and stable for up to 50 days. In conclusion, this novel formulation increases the stability/robustness of ADI, eases the product separation process and could be reused for economically viable bioproduction of nutraceutical citrulline in industries. [Display omitted] • Novel cross-linked enzyme aggregates of arginine deiminase (ADI-CLEA) were reported. • Immobilization conditions were optimized by network-based multiobjective genetic algorithm. • ADI-CLEA exhibited enhanced catalytic efficiency, stability and robustness. • Improved citrulline yield (215 g/L) was obtained by ADI-CLEA. • ADI-CLEA could be recycled for seven cycles without losing its activity. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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