Carbon Sink Potential of Sulfur-Oxidizing Bacteria in Groundwater at Petroleum-Contaminated Sites

Groundwater at petroleum-contaminated sites typically exhibits elevated dissolved inorganic carbon (DIC) levels due to hydrocarbon biodegradation; however, our prior field investigations revealed an enigmatic DIC depletion anomaly that starkly contradicts this global pattern and points to an unrecognized carbon sink. In a breakthrough demonstration, this study provides the first experimental confirmation that sulfur-oxidizing bacteria (SOB) drive substantial carbon sequestration via a coupled sulfur oxidation autotrophic assimilation process. Through integrated hydrochemical monitoring and 16S rRNA sequencing in an enrichment culture system, we captured the complete DIC transformation trajectory: heterotrophic acetate degradation initially increased DIC to 370 mg/L, but subsequent autotrophic assimilation by SOB dramatically reduced DIC to 270 mg/L, yielding a net consumption of 85 mg/L. The distinctive pH dynamics (initial alkalization followed by acidification) further corroborated microbial regulation of carbon cycling. Critically, Pseudomonas stutzeri and P. alcaliphila were identified as the dominant carbon-fixing agents. These findings definitively establish that chemolithoautotrophic SOB convert DIC into organic carbon through a "sulfur oxidation-carbon fixation" coupling mechanism, overturning the conventional paradigm of petroleum-contaminated sites as perpetual carbon sources. The study fundamentally redefines natural attenuation frameworks by introducing microbial carbon sink potential as an essential assessment metric for environmental sustainability.