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Turnover Number and Catalytic Efficiency

Turnover Number and Catalytic Efficiency

The turnover number of an enzyme is the maximum number of substrate molecules it can transform per unit time. Turnover numbers for most enzymes range from 1 to 1000 molecules per second. Catalase has the known highest turnover number, capable of converting up to 2.8×106 molecules of hydrogen peroxide into water and oxygen per second. Lysozyme has the lowest known turnover number of half a molecule per second.
Chymotrypsin is a pancreatic enzyme that breaks down proteins during digestion....

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Long-Term Biocatalyst Performance: Mechanistic Prediction and Continuous Non-Isothermal Testing.

Thomas A Rogers^1,2,3, Luis G Encarnación-Gómez¹, Andreas S Bommarius^1,2,4

¹School of Chemical & Biomolecular Engineering, Georgia Institute of Technology, Atlanta, Georgia, 30332, USA.

|April 20, 2022

Summary

Two new methods estimate biocatalyst stability faster and cheaper than traditional tests. These approaches accurately predict total turnover number (TTN), crucial for assessing enzyme productivity and lifetime.

Keywords:

TEM-1 β-lactamase lifetime productivity non-isothermal kinetics operational stability total turnover number

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Area of Science:

Biocatalysis
Enzyme Kinetics
Chemical Engineering

Background:

Assessing biocatalyst operational stability via total turnover number (TTN) using conventional isothermal experiments is time-consuming and resource-intensive.
Stable biocatalysts present particular challenges for accurate TTN determination due to slow deactivation rates.

Purpose of the Study:

To develop and validate alternative methods for estimating the TTN of biocatalysts.
To provide faster and more efficient alternatives to conventional TTN assessment methods.

Main Methods:

A mechanistic approach utilizing enzyme deactivation models derived from first principles, requiring two isothermal batch measurements.
A non-isothermal continuous-mode approach using limited experiments and mathematical modeling to identify intrinsic deactivation parameters.

Main Results:

Both developed methods provide TTN estimates within a factor of two to five of values obtained from traditional, lengthy isothermal aging tests.
Validation was performed using TEM-1 β-lactamase-catalyzed penicillin G hydrolysis.

Conclusions:

The mechanistic and non-isothermal continuous methods offer significant advantages over conventional TTN determination.
These alternative approaches are valuable for calculating catalyst cost in continuous processing and for efficient screening of biocatalysts for stability.