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. 2010 Mar 15;65(6):2118-2124.
doi: 10.1016/j.ces.2009.12.005.

Utilizing Simple Biochemical Measurements to Predict Lifetime Output of Biocatalysts in Continuous Isothermal Processes

Thomas A Rogers  1 , Andreas S Bommarius
Affiliations

Utilizing Simple Biochemical Measurements to Predict Lifetime Output of Biocatalysts in Continuous Isothermal Processes

Thomas A Rogers et al. Chem Eng Sci. .

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.

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Figures

Scheme 1
Scheme 1
Lumry-Eyring Model (Lumry and Eyring 1954)
Scheme 2
Scheme 2
Equilibrium Model (Daniel et al., 2001)
Scheme 3
Scheme 3
Extended Lumry-Eyring Model (Gibbs et al., 2005)
Scheme 4
Scheme 4
Multiple-Intermediate Model (Freire et al., 1990)
Scheme 5
Scheme 5
Multiple Intermediate Model with Direct Denaturation Pathways
See this image and copyright information in PMC

References

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