To comprehensively identify acyl-CoA binding protein (ACBP) genes in the Chinese gutta percha tree, we utilized known ACBP sequences from Arabidopsis, rice, and poplar as references, employing blastp, hmmer, and domain analysis techniques. This led to the identification of five distinct ACBPs (EuACBP1-EuACBP5) characterized by varying amino acid content and molecular weights, all of which are acidic with an instability index suggesting inherent instability. Chromosomal mapping revealed these genes on chromosomes 7 and 8, with a uniform distribution across different plant species' chromosomes. Exon-intron organization analysis indicated a consistent structure within subgroups, reflecting potential functional significance and evolutionary patterns. Predicted subcellular localizations showed EuACBP1, EuACBP2, and EuACBP4 in the cytoplasm, while EuACBP3 and EuACBP5 were in the chloroplasts and nucleus, respectively. Comparative phylogenetic analysis categorized EuACBPs into four subgroups, highlighting the evolutionary relationships with homologous genes from Arabidopsis, rice, and poplar. Cis-acting element analysis of promoter regions identified motifs related to stress responses, hormone response, and plant growth, suggesting regulatory roles in various physiological processes. Tissue-specific expression analysis demonstrated distinctive patterns, with EuACBP1 predominantly in seeds and EuACBP5 in roots and stems, indicating their involvement in specific biological functions. Seasonal and drought stress expression analysis revealed significant seasonal specificity and varied responses to drought stress, emphasizing their roles in plant adaptation. This study enriches our understanding of the ACBP gene family in the Chinese gutta percha tree, providing a foundation for future research into their functional dynamics under environmental stresses. Comprehensive gene interaction network, miRNA regulatory effects, and KEGG pathway enrichment analyses further highlighted their crucial roles in metabolism, genetic information processing, and cellular processes. These findings underscore the potential genetic targets for enhancing stress resistance and adaptability in plants. This study comprehensively identified and characterized five ACBP genes in the Chinese gutta percha tree, revealing their distinct amino acid content, subcellular localizations, and chromosomal distributions. Phylogenetic analysis categorized these genes into four subgroups, highlighting evolutionary relationships. Promoter analysis uncovered cis-elements related to stress and hormonal responses, while tissue-specific and seasonal expression patterns indicated their roles in various biological functions and stress responses. These findings enhance our understanding of the ACBP gene family, providing a foundation for future research on their functional dynamics and potential applications in improving stress resistance and adaptability in plants.