A study on the refolding of lysozyme using fed-batch and step-addition strategies

摘要

The refolding step is essential for industrially produced protein aggregates to restore their biological activities. Various strategies, including the direct dilution method, have been utilized to enhance the performance of the refolding process; however, refinement and improvement on these methods are ongoing. We have used two different operating schemes, fed-batch and step-addition, independently and collaboratively to refold denatured lysozymes. Our research shows that the fed-batch and the step-addition schemes overcame the aggregation formation and favored the refolding step at higher feeding concentrations in comparison with the typical dilution method. Moreover, with relatively longer folding time intervals, the step-addition scheme provided higher activity recovery, while the fed-batch scheme was advantageous in terms of productivity. Finally, a combined scheme was proposed to optimize the refolding process. This work demonstrates the feasibility of applying fed-batch and/or step-addition operation schemes to the renaturation of enzyme activity.