Hexanitrohexaazaisowurtzitane (CL-20) is a compound with a polycyclic cage and an N-nitro group that has been shown to play an unfavorable role in environmental fate, biosafety, and physical health. The aim of this study was to isolate the microbial community and to identify a single microbial strain that can degrade CL-20 with desirable efficiency. Metagenomic sequencing methods were performed to investigate the dynamic changes in the composition of the community diversity. The most varied genus among the microbial community was Pseudomonas, which increased from 1.46% to 44.63% during the period of incubation (MC0–MC4). Furthermore, the new strain was isolated and identified from the activated sludge by bacterial morphological and 16s rRNA sequencing analyses. The CL-20 concentrations decreased by 75.21 μg/mL and 74.02 μg/mL in 48 h by MC4 and Pseudomonas sp. ZyL-01, respectively. Moreover, ZyL-01 could decompose 98% CL-20 of the real effluent in 14 day’s incubation with the glucose as carbon source. Finally, a draft genome sequence was obtained to predict possible degrading enzymes involved in the biodegradation of CL-20. Specifically, 330 genes that are involved in energy production and conversion were annotated by Gene Ontology functional enrichment analysis, and some of these candidates may encode enzymes that are responsible for CL-20 degradation. In summary, our studies indicate that microbes might be a valuable biological resource for the treatment of environmental contamination caused by CL-20 and that Pseudomonas sp. ZyL-01 might be a promising candidate for eradicating CL-20 to achieve a more biosafe environment and improve public health.

六硝基六氮杂异武兹烷 (CL-20) 是一种具有多环笼和 N-硝基的化合物，已被证明对环境命运、生物安全和身体健康起着不利的作用。本研究的目的是分离微生物群落并鉴定能够以理想效率降解 CL-20 的单一微生物菌株。采用宏基因组测序方法研究群落多样性组成的动态变化。微生物群落中变化最大的属是假单胞菌属，在培养期间（MC0-MC4）从 1.46% 增加到 44.63%。此外，通过细菌形态学和16s rRNA测序分析从活性污泥中分离和鉴定了新菌株。 MC4 和假单胞菌在 48 小时内使 CL-20 浓度降低了 75.21 μg/mL 和 74.02 μg/mL。分别为ZyL-01。此外，ZyL-01以葡萄糖为碳源，经过14天的培养，可以分解实际废水中98%的CL-20。最后，获得了基因组序列草图，以预测参与 CL-20 生物降解的可能的降解酶。具体来说，通过基因本体功能富集分析注释了涉及能量产生和转换的330个基因，其中一些候选基因可能编码负责CL-20降解的酶。总之，我们的研究表明微生物可能是治疗 CL-20 和假单胞菌引起的环境污染的宝贵生物资源。 ZyL-01 可能是消除 CL-20 的有前途的候选药物，以实现更加生物安全的环境并改善公众健康。