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放线菌红色糖多孢菌 CGMCC 17550 通过一种新的羟化途径降解新烟碱类杀虫剂噻虫嗪,并修复地表水中的噻虫嗪。

Actinomycetes Rhodococcus ruber CGMCC 17550 degrades neonicotinoid insecticide nitenpyram via a novel hydroxylation pathway and remediates nitenpyram in surface water.

机构信息

Jiangsu Key Laboratory for Microbes and Functional Genomics, Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources, College of Life Science, Nanjing Normal University, Nanjing, 210023, People's Republic of China.

出版信息

Chemosphere. 2021 May;270:128670. doi: 10.1016/j.chemosphere.2020.128670. Epub 2020 Oct 17.

Abstract

Neonicotinoid insecticides are neurotoxicants that cause serious environmental pollution and ecosystem risks. In the present study, a nitenpyram-degrading bacterium, Rhodococcus ruber CGMCC 17550, was isolated from a nitenpyram production sewage treatment tank. Liquid chromatography-mass spectrometry analysis revealed R. ruber degraded nitenpyram via a novel hydroxylation pathway to form three different metabolites, one of which was confirmed to hydroxylate nitenpyram at the C3 site of the 6-chlorpyridine cycle by nuclear magnetic resonance analysis. The nitenpyram degradation rate increased as the biomass of resting R. ruber CGMCC 17550 cells increased, reaching 98.37% at an OD of 9 in transformation broth containing 100 mg L nitenpyram after 72 h of incubation. Nitenpyram degradation by R. ruber CGMCC 17550 was insensitive to dissolved oxygen levels. Use of glucose, fructose and pyruvate as co-substrates slightly increased nitenpyram degradation. The cytochrome P450 inhibitor 1-aminobenzotriazole strongly inhibited nitenpyram degradation, indicating that P450 enzymes may mediate nitenpyram hydroxylation. Inoculation of R. ruber CGMCC 17550 enhanced nitenpyram degradation in surface water. Additionally, R. ruber cells immobilized by calcium-alginate remediated 87.11% of 100 mg L NIT in 8 d. Genome sequencing analysis confirmed that R. ruber CGMCC 17550 has metabolic diversity and abundant KEGG genes involved in xenobiotics biodegradation and metabolism. These findings demonstrate that R. ruber CGMCC 17550 is capable of unique biodegradation of nitenpyram via the hydroxylation pathway and is a promising bacterium for bioremediation of contaminants.

摘要

新烟碱类杀虫剂是神经毒物,会造成严重的环境污染和生态风险。本研究从吡虫啉生产污水处理罐中分离到一株能够降解吡虫啉的诺卡氏菌 Rhodococcus ruber CGMCC 17550。液质联用分析表明,R. ruber 通过一种新的羟化途径降解吡虫啉,生成 3 种不同的代谢产物,其中一种经核磁共振分析确认为在 6-氯吡啶环的 C3 位羟化吡虫啉。当休止细胞的生物量增加时,R. ruber CGMCC 17550 对吡虫啉的降解速率增加,在转化液中含 100mg/L 吡虫啉,OD 值为 9,培养 72h 后,吡虫啉的降解率达到 98.37%。R. ruber CGMCC 17550 对溶解氧水平不敏感。以葡萄糖、果糖和丙酮酸作为共底物略微提高了吡虫啉的降解率。细胞色素 P450 抑制剂 1-氨基苯并三唑强烈抑制吡虫啉的降解,表明 P450 酶可能介导吡虫啉的羟化。接种 Rhodococcus ruber CGMCC 17550 增强了地表水中吡虫啉的降解。此外,用海藻酸钙固定化 Rhodococcus ruber 细胞在 8d 内修复了 100mg/L NIT 的 87.11%。基因组测序分析证实,R. ruber CGMCC 17550 具有代谢多样性,并且有丰富的与外源性化合物生物降解和代谢相关的 KEGG 基因。这些结果表明,R. ruber CGMCC 17550 能够通过羟化途径对吡虫啉进行独特的生物降解,是一种有前途的生物修复污染物的细菌。

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