Experimental and computational datasets for signal peptidase I from Candidatus Liberibacter asiaticus

This dataset supports the study of signal peptidase I from Candidatus Liberibacter asiaticus, a membrane-associated enzyme that is difficult-to-produce in a functional form. The objective of this work was to evaluate and compare multiple recovery and purification strategies to obtain soluble and active protein.

The data were generated through a combination of experimental and computational approaches to evaluate multiple recovery strategies, including refolding from inclusion bodies, detergent-based extraction of the full-length protein, and purification using affinity tags and chromatographic techniques.

The dataset includes fluorescence-based activity measurements (FRET assays), chromatographic profiles from purification steps, structural models generated in Schrodinger Maestro, binding energy calculations obtained from MMGBSA analysis, and assessments of protein purity. FRET assay data provide fluorescence intensity readings used to evaluate enzymatic activity under different conditions. Chromatographic data include absorbance profiles (A280 and A350) used to assess protein recovery and purification performance. Structural and computational datasets include protein models and ligand complexes used for docking and energetic analysis.

The results show that high protein recovery does not necessarily correlate with enzymatic activity. In several cases, proteins recovered in the soluble fraction did not exhibit detectable activity, suggesting the presence of soluble aggregates or improperly folded species. Detergent-based extraction of the full-length protein was ineffective under the conditions tested, and while affinity tags improved recovery, they did not prevent aggregation or restore functionality.

These data should be interpreted in the context of evaluating the relationship between protein recovery, aggregation, and functionality. The dataset can be used to compare different recovery strategies, assess purification performance, and support further analysis of aggregation-prone membrane proteins. All files are organized by method and provided in standard formats.