Multivariate curve resolution for analysis of Raman hyperspectral imaging data sets for enzyme immobilization

• Raman hyperspectral imaging of an enzyme immobilized onto two resins was accomplished. • Multivariate data analysis spectrally resolved chemical species involved in immobilization. • Chemical images spatially resolved chemical species involved in immobilization. • The Raman hyperspectral imaging data sets can be used for alternative applications. Biocatalytic processes are an essential component in achieving efficient and productive chemical syntheses in the scientific and industrial communities. Immobilization of the biocatalyst offers distinct advantages for the process, including increasing the stability and reusability of the biocatalyst as well as increasing control over the process. Herein, directly engineered pantothenate kinase (PanK) was immobilized to two commercially available microporous resins. Ralbovsky et al. (2021) utilized Raman hyperspectral imaging in combination with multivariate curve resolution alternating least squares (MCR-ALS) to spatially and spectrally resolve the chemical species involved in the enzyme immobilization process. Here, we provide eight Raman hyperspectral imaging data sets collected from the enzyme immobilized to the resin beads under different collection parameters, encompassing over 26,000 raw spectra. A brief discussion of the results, in the form of chemical maps and chemical images, indicates both resins, PanK, the glass substrate, and a Bis-Tris wash solution were resolved using MCR-ALS analysis of a subset of hyperspectral images, thus providing crucial chemical and spatial information about the system. The eight total Raman hyperspectral imaging data sets are shared to provide the scientific community with a thoroughly characterized set of spectral data which can be used for a variety of applications, including for further understanding and optimization of enzyme immobilization systems as well as for individual learning and development applications in the fields of chemical imaging and chemometrics.