[Aim]  To determine the physicochemical properties and molecular characteristics of the quinate permease gene (qutD) in the fungus Ascosphaera apis (AaqutD) and thereby provide a foundation for in-depth investigation of the function of this gene. [Methods]  The coding sequence (CDS) of AaqutD underwent PCR amplification and subsequent TA cloning. Predictions regarding its physicochemical properties, signal peptide, phosphorylation sites, secondary and tertiary structures, as well as protein interaction network, were made utilizing appropriate software tools. Conserved motifs and domains of this gene in As. apis and ten other fungi were forecasted through MEME software and the BatchCD-Search tool, respectively. Subsequently, a phylogenetic tree was constructed using Mega11.0 software. The relative expression of AaqutD in As. apis infecting worker larvae of Ap. cerana cerana was assessed using RT-qPCR. [Results]  The AaqutD CDS was successfully cloned. AaqutD was expressed in As. apis spores. The molecular weight of AaqutD is around 59.66 kD, with a molecular formula of C₂₇₆₂H₄₁₄₉N₆₈₉O₇₃₃S₂₉. It has a fat coefficient of 88.77, an isoelectric point of 7.51, and an average hydrophilicity coefficient of 0.286. AaqutD was simultaneously localized in the endoplasmic reticulum, mitochondria and nuclei. AaqutD has 40 phosphorylation sites, 240 α-helices, 111 extended chains, 29 β-turns and 155 irregular coils, however, it lacks a typical signal peptide. There is 100% sequence homology between AaqutD and the template A0A168DJE0.1.A. AaqutD may interact with 10 proteins including the tumor protein p73. The same structural domain (Sugartr) and the same eleven conserved motifs, were identified in qutD in As. apis and 10 other fungi. qutD of As. apis and Ap. acerosa clustered in one clade on the phylogenetic tree. Expression of AaqutD was significantly upregulated (P<0.05) 2- and 3-days post-inoculation (d) compared to that at 1 d. [Conclusion]  AaqutD is a potential hydrophilic and intracellular protein that is highly conserved and homologous with the qutD gene of other fungi. AaqutD is most closely related to the qutD gene of Ap. acerosa. AaqutD is significantly activated in As. apis during the infection of Ap. cerana cerana worker larvae.