菌株Pseudomonas.BTs降解转化3种苯并三唑类化合物

Biotransformation of Three Benzotriazoles with Bacterial Strain Pseudomonas.BTs

  • 摘要: 在好氧条件下分离获得一株苯并三唑类化合物(Benzotriazoles, BTs)降解菌, 通过16S rDNA测序和数据库比对分析显示与Pseudomonas.taiwanensis BCRC 17751同源性最高, 并将该菌株命名为Pseudomonas.BTs.该菌株在外加碳源存在时能够以不同的速率降解3种典型BTs(苯并三唑, BTri; 5-甲基苯并三唑, 5-TTri; 5-氯-苯并三唑, CBT), 但无法以BTs为唯一碳源.测试了Pseudomonas.BTs利用11种外加碳源作为生长基质共代谢BTs的效果.结果表明:外加碳源投加质量比(mC:mBTs)=1 000:1时比100:1的情况更有利于BTs的共代谢.在11种外碳源中, 葡萄糖、谷氨酸钠和乙醇最有利于Pseudomonas.BTs共代谢BTs, 而当以苯酚、麦芽糖、淀粉作为外碳源时, BTs的共代谢完全没有发生.利用超高效液相色谱与四极杆飞行时间高分辨率质谱联用技术鉴定了Pseudomonas.BTs转化BTri、5-TTri和CBT的产物.结果表明:3种BTs化合物具有相对一致的转化路径, 包括异构化、甲基化、甲氧基化以及其它官能团的加成反应.多数产物为首次被发现.研究结果可以为探索微生物转化BTs的机理、优化去除BTs的研究提供参考.

     

    Abstract: A pure Benzotriazoles (BTs) degrading culture isolated under aerobic condition was investigated. Analysis of the 16S rDNA sequence and database indicated that the closest strain was the Pseudomonas.taiwanensis BCRC 17751, and this isolated strain was named as Pseudomonas.BTs. The isolated strain was able to degrade three BTs (BTri, 5-TTri and CBT) with different removal rates in the presence of external carbon source but could not utilize BTs as sole carbon source. The effects of 11 external carbon sources used as growth substrate on the co-metabolism of BTs by Pseudomonas.BTs were tested. The feed mass ratio (mC:mBTs) of 1 000:1 would be better for co-metabolism of BTs than 100:1. Among all 11 substances, glucose, sodium glutamate and ethanol were the most beneficial to co-metabolism of BTs by Pseudomonas.BTs, whereas the co-metabolism of BTs did not happen at all when phenol, maltose, humic acid or starch served as the source of external carbon. The biotransformation products (TPs) of BTri, 5-TTri and CBT by Pseudomonas.BTs were identified using ultra high-performance liquid chromatography coupled with a quadrupole-time-of-flight high-resolution mass spectrometer (UHPLC-QToF-MS). The results indicated that the three BTs shared relatively consistent transformation pathways including isomerization, methylation and other similar functional group addition reactions. Most TPs were reported for the first time. The results obtained in this experiment may provide basic information for understanding the mechanism of biotransformation of BTs as well as the optimization of BTs removal.

     

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