<p>We previously developed a spherical hydrogel containing β-cyclodextrin (β-CyD) for the recovery of phenol (PhOH) from PhOH-containing wastewater by exploiting the inclusion capability of β-CyD toward aromatic compounds. In this study, a mixed-type spherical polyvinyl alcohol (PVA) hydrogel (PVA<i>-mix-</i>βCyD/hexamethylene diisocyanate (HDI)) was prepared, in which βCyD/HDI was accumulated and distributed at concentrations ranging from 18–55 μmol g⁻¹. The hydrogels had a diameter of ~3 mm and a moisture content of 79–70 wt%. Scanning electron microscopy revealed surface internetwork distances of 20 μm for the hydrogel without β-CyD and 20 μm (18–21 μmol g⁻¹), 25 μm (34 μmol g⁻¹), and 30 μm (36–55 μmol g⁻¹) for the PVA<i>-mix-</i>βCyD/HDI hydrogels. In contrast to that of the PVA<i>-co-</i>βCyD hydrogels, where β-CyD was uniformly dispersed, network spacing increased proportionally with increasing β-CyD content. PhOH-degrading bacteria were immobilized as biocatalysts and tested in model wastewater (500 mg L⁻¹ PhOH). The degradation rate of the β-CyD-containing biocatalyst was approximately twice that of the β-CyD-free biocatalyst, with saturation above 20 μmol g⁻¹. The β-CyD content at saturation was lower for the PVA<i>-mix-</i>βCyD/HDI biocatalyst than for the PVA<i>-co-</i>βCyD biocatalyst. Furthermore, fluorescence in situ hybridization confirmed uniform bacterial dispersion throughout the inner layer of the gel, with the bacterial abundance increasing as the β-CyD content increased.</p>