Heavy metal contamination from abandoned mines presents long-term risks to soil ecosystems by altering physicochemical conditions and limiting microbial functions. To investigate these effects, we analyzed soils from the Deoksan Pb-Zn abandoned mine in Korea using community-level physiological profiling (CLPP) with Biolog EcoPlateTM. Soil samples were collected from three contaminated sites and one uncontaminated control, and we assessed their physicochemical properties, heavy metal concentrations, and microbial substrate utilization patterns over a 7-day incubation period. The results revealed significant site-specific differences in soil chemistry, with Zn and Pb concentrations exceeding ecological safety thresholds near the mine adit. Average well color development (AWCD) increased over time across all sites, but functional trajectories differed: highly contaminated soils exhibited prolonged increases, while low-contamination soils plateaued earlier. Substrate utilization patterns shifted over time, with carbohydrates and carboxylic acids dominating in the early incubation phase, while phosphorylated chemicals became more prominent in later stages. Multiple regression and relative importance analyses identified Cd, Pb, and Zn as key regulators of substrate utilization, with phosphorylated chemicals showing strong negative correlations (R 2>0.95). These findings indicate that heavy metal stress not only decreases overall microbial activity but also disrupts specific metabolic pathways. The utilization of phosphorylated chemicals emerged as a particularly sensitive functional indicator, underscoring its potential for ecological risk assessment and soil health monitoring in contaminated sites.

Abstract
1. INTRODUCTION
2. MATERIALS AND METHODS
    2.1. Study site and selection ofsampling locations
    2.2. Soil sampling and analysis
    2.3. Microbial community substrateutilization profiles
    2.4. Statistical analysis
3. RESULTS
    3.1. Soil physicochemical properties andheavy metal concentrations
    3.2. Temporal changes in substrate utilization
    3.3. Site- and time-dependent variation insubstrate group utilization
    3.4. Influence of heavy metal concentrationson substrate utilization
4. DISCUSSION
    4.1. Influence of soil properties and heavymetals on microbial functional diversity
    4.2. Temporal dynamics of substrateutilization
    4.3. Shifts in substrate group utilizationunder metal stress
    4.4. Methodological considerations andtemporal variability
    4.5. Implications for ecological riskassessment and soil health monitoring
5. CONCLUSION
CRediT authorship contribution statement
Declaration of Competing Interest
ACKNOWLEDGMENTS
REFERENCES

• Minseok Park(OJeong Resilience Institute, Korea University, Seoul 02841, Republic of Korea)
• Yongeun Kim(OJeong Resilience Institute, Korea University, Seoul 02841, Republic of Korea)
• Yongho Lee(OJeong Resilience Institute, Korea University, Seoul 02841, Republic of Korea, Humanities and Ecology Consensus Resilience Laboratory, Hankyong National University, Anseong 17579, Republic of Korea)
• Dokyung Kim(Department of Environmental Health Science, Konkuk University, Seoul 05029, Republic of Korea)
• Youn-Joo An(Department of Environmental Health Science, Konkuk University, Seoul 05029, Republic of Korea)
• Seunghun Hyun(Department of Environmental Science and Ecological Engineering, Korea University, Seoul 02841, Republic of Korea)
• Sun Hee Hong(School of Plant Science and Landscape Architecture, Hankyong National University, Anseong 17579, Republic of Korea)
• June Wee(Department of Applied Biology, Chungnam National University, Daejeon 34143, Republic of Korea) Corresponding author