Hybrid strategy for the selective recovery of the intracellular enzyme from the cytoplasmic location using hydrodynamic cavitation: Scalable approach

Abstract

Hydrodynamic cavitation technology is a promising method for the selective extraction of intracellular enzymes. But the scalability of the method is the major challenge for its large-scale operations. In the present work, we have studied a scalable approach for the selective recovery of alcohol dehydrogenase (ADH) enzyme from the cytoplasm of yeast cells has been proposed. The scalable hybrid strategy involves the pH pretreatment for translocation of the cytoplasmic enzyme to the periplasmic location, followed by enzyme recovery using hydrodynamic cavitation (orifice device, 0.2mm orifice diameter). The hybrid strategy of pH pretreatment (10L, 8pH, 12h) followed by the hydrodynamic cavitation-assisted cell disruption (10L, 6 bar, 5 passes) increases the selective release (12.66 U/mg, 6 bar, 5 passes) by 8.22 times and the specific yield (344.46 U/kJ, 6 bar, 5 passes) by11.98 times as was observed when it is compared with hydrodynamic cavitation assisted cell disruption without pre-treatment (selectivity of 1.54 U/mg and specific yield of 28.76 U/kJ at 6 bar, 5 passes).