The objective of this work was to evaluate the effect of influent pH on the hydrolytic acidification (HA) performance and microbial community structure in an expanded granular sludge bed (EGSB) pretreating crotonaldehyde manufacture wastewater (CMW) after ozonation. The results showed that higher chemical oxygen demand (COD) removal rate (40.1%) and acidification degree (27.6%) were obtained at pH 8.0 than those at pH 6.0 and pH 4.0. The concentration of extractable extracellular polymeric substance (EPS) in the sludge gradually decreased with the pH decreasing from 8.0 to 4.0. A similar change was also observed for the concentration of total volatile fatty acids (TVFA) in the effluent. The optimal detoxification efficiency by the HA process was obtained at pH 8.0, with higher removal efficiency (all higher than 90%) of the main toxic pollutants (crotonaldehyde, 5-formyl-6-methyl-4,5-dihydropyran, etc.) and higher anaerobic biodegradation rate (44.5%) in biochemical methane potential (BMP) assay. Among the predominant genera, the Acinetobacter and Pseudomonas were possibly related to biodegradation of pollutants, since their higher relative abundance also coincided with the better performance of the HA process at pH 8.0.