Utilizing simple biochemical measurements to predict lifetime output of biocatalysts in continuous isothermal processes

Abstract

The expected product yield of a biocatalyst during its useful lifetime is an important consideration when designing a continuous biocatalytic process. One important indicator of lifetime biocatalyst productivity is the dimensionless total turnover number ( TTN). Here, a method is proposed for estimating the TTN of a given biocatalyst from readily measured biochemical quantities, namely the specific activity and the deactivation half-life, measured under identical conditions. We demonstrate that this method may be applied to any enzyme whose thermal deactivation follows first-order kinetics, regardless of the number of unfolding intermediates, and that the TTN method circumvents the potential problems associated with measuring specific catalyst output when a portion of the enzyme is already unfolded. The TTN estimation was applied to several representative biocatalysts to demonstrate its applicability in identifying the most cost-effective catalyst from a pool of engineered mutants with similar activity and thermal stability.