TY - JOUR AU - Shafi, Azra AU - Husain, Qayyum PY - 2022 DA - 2022/11/17 TI - Effect of Different Additives on the Structure and Activity of β-Galactosidase Immobilized on a Concanavalin A–Modified Silica-Coated Titanium Dioxide Nanocomposite JO - Catalysis Research SP - 040 VL - 02 IS - 04 AB - Interpreting the relationship between the activity and structure of β-galactosidase is necessary to perceive the impact of the enzyme’s conformation on its catalysis. The current study thoroughly explains the effects of additives such as ethylenediaminetetraacetic acid (EDTA), sodium dodecyl sulfate (SDS), dithiothreitol (DTT), and urea on β-galactosidase activity and structure. β-Galactosidase activity was determined at various ionic strengths and temperatures as a function of time. Structural studies evaluating changes in the secondary and tertiary structures of the enzyme in the presence of the additives were conducted using ultraviolet (UV)-visible and intrinsic fluorescence spectroscopy. The immobilized enzyme showed enhanced stability under different environmental conditions. Activity assays demonstrated concentration-dependent inactivation of β-galactosidase in the presence of SDS and urea, which suggests that hydrophobic and charged residues are present near the active site. In the presence of EDTA, loss in activity was noted, which confirms that β-galactosidase is a metalloenzyme. Enhancement in enzyme activity in the presence of DTT suggests the presence of a cysteine residue near the catalytic center. In UV-visible and intrinsic fluorescence spectroscopy studies, the native enzyme showed significant conformational transitions in the presence of DTT, SDS, and urea and very few changes in the presence of EDTA. However, the immobilized enzyme could resist significant structural changes. In conclusion, this study provides a detailed description of the association between the activity and conformational stability of β-galactosidase. SN - 2771-490X UR - https://doi.org/10.21926/cr.2204040 DO - 10.21926/cr.2204040 ID - Shafi2022 ER -