Whole-Cell Biocatalytic Degradation of Heavy Oil Under Low Temperature by a Pseudomonas Strain Isolated from Oil-Contaminated Sites

The removal of heavy oil under low-temperature conditions is a significant global challenge. This study aimed to assess the long-term whole-cell biocatalytic degradation of heavy oil in water and soil by bacteria isolated from contaminated soil in Muroran, Japan, under cold conditions. Enrichment cultures using heavy oil as the sole carbon source yielded 15 potent heavy oil-degrading isolates. However, only the C1 strain retained its activity under low-temperature conditions and was identified as Pseudomonas aeruginosa C1 using 16S rDNA sequencing. Gas chromatography analysis revealed that at 30 °C (water medium), strain C1 degraded 57% of heavy oil within 7 days. At 15 °C, the degradation efficiency of C1 declined due to a temperature-dependent metabolic lag phase (1 day); however, at 15 °C, 70% degradation was observed in seven days. In long-term experiments at 5 °C and 10 °C, 35% and 40% degradation were recorded for C1 after 98 days. In artificially contaminated soil at 5 °C, C1 achieved 60% biodegradation. These results demonstrate cold-adapted whole-cell activity against heavy oil and motivate the design of controlled, contained ex situ reactors (e.g., enzyme-based or cell-free systems) for safe remediation in cold climates.