Accurate modeling of enzyme activity and stability is an important goal of the protein engineering community. However, studies seeking to evaluate current progress are limited by small data sets of quantitative kinetic constants and thermal stability measurements. Here, we report quantitative measurements of soluble protein expression in E. coli, thermal stability, and Michaelis-Menten constants (kcat, KM, and kcat/KM) for 129 designed mutants of a glycoside hydrolase. Statistical analyses reveal that functional Tm is independent of kcat, KM, and kcat/KM in this system, illustrating that an individual mutation can modulate these functional parameters independently. In addition, this data set is used to evaluate computational predictions of protein stability using the established Rosetta and FoldX algorithms. Predictions for both are found to correlate only weakly with experimental measurements, suggesting improvements are needed in the underlying algorithms.
Submitter: Shu-Ching Ou
Submission Date: Oct. 10, 2018, 12:31 p.m.
|Number of data points||1087|
|Assays/Quantities/Protocols||Experimental Assay: Kinetic Constant: Km ; Experimental Assay: Kinetic Constant: kcat ; Experimental Assay: Thermal Stability: Tm ; Experimental Assay: Expression/Solubility ; Derived Quantity: SD of Kinetic Constant: kcat/Km ; Derived Quantity: Kinetic Constant: kcat/Km ; Derived Quantity: SD of Kinetic Constant: Km ; Derived Quantity: SD of Kinetic Constant: kcat ; Derived Quantity: SD of Thermal Stability: Tm|
|Libraries||Enzyme functional parameters for mutants of BglB|
|Percent Identity||Matching Chains||Protein||Accession||Entry Name|