The reduction of hexavalent chromium [Cr(VI)] by microbiota agents is a potential method for ecological remediation in heavily contaminated soil. This study investigated the contribution of microbial community and extracellular polymeric substances (EPS) to reduce Cr(VI) using lab-cultured anaerobic granular sludge (AGS). The AGS demonstrated an impressive reduction efficiency of 99.3% for Cr(VI) in soil, operating within an initial Cr(VI) concentration range of 500–3000 mg/kg. The distribution of typical electron acceptors, e.g., SO42-, NO3-, and Fe3+, had a negligible influence on microbial Cr(VI) reduction. The contribution of EPS on Cr(VI) reduction accounted for only 2.1% of the total Cr(VI) reduction, implying that Cr(VI) reduction driven by AGS was primarily caused by the direct effect of microbiota. High-throughput sequencing analysis confirmed that an increasing trend in the relative abundance of Cr reduction bacteria (Sphingopyxis: 0.40–5.54%, Thermonas: 0.41–5.70%, Edaphobaculum: 0.35–6.64%, etc.), and the stability (0.716–4.114) of microbial co-occurrence networks was significantly promoted by the end of the experiment. Materials characterization indicated that successful reduction of Cr(VI) to trivalent chromium [Cr(III)] and formed in Cr(OH)3, and functional groups, O-H, N-H, -CH, and -CH2, of AGS likely played a vital role in Cr(VI) reduction. Overall, these findings offer insights into the remediation technique of microbial Cr(VI) reduction in highly-contaminated soil based on the interaction network stability of the microbial community in AGS.