The explosive and biorefractory nature of nitrocellulose (NC) poses major risks to both humans and the environment. Expanding the range of microorganisms capable of degrading NC is essential, though the most effective known microorganisms, Desulfovibrio genera and Fusarium solani, achieve degradation rates of 5%-25%. Here, a novel strain, Rhodococcus pyridinivorans LZ1 was isolated, demonstrating the ability to degrade NC, with its growth potentially enhanced by the presence of NC. The degradation process was monitored by assessing changes in nitrate, nitrite, and ammonium. Notably, the –OH strength of NC increased over time, whereas the energetic functional groups (–NO2 and O-NO2) diminished. Furthermore, the presence of NC enhanced nitrate esterase activity 1–2-fold, indicating that ammonification was the primary pathway for NC biodegradation. By converting the nitrate ester of NC into hydroxyl, R. pyridinivorans LZ1 mitigates the harmful effects of NC, offering a promising approach for the treatment of NC waste and wastewater.