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一株内生菌 Aeromonas hydrophila S27 对孔雀石绿的生物降解及其新型氧化还原酶的参与。

Biodegradation of malachite green by an endophytic bacterium Klebsiella aerogenes S27 involving a novel oxidoreductase.

机构信息

College of Life Science, Shandong Normal University, Jinan, 250014, People's Republic of China.

College of Biological Science and Technology, Hunan Agricultural University, Changsha, 410128, People's Republic of China.

出版信息

Appl Microbiol Biotechnol. 2019 Mar;103(5):2141-2153. doi: 10.1007/s00253-018-09583-0. Epub 2019 Jan 6.

Abstract

Endophytic microorganisms can metabolize organic contaminants and assist in plant growth, thus facilitating the phytoremediation of polluted environments. An endophytic bacterium capable of decoloring malachite green (MG) was isolated from the leaves of the wetland plant Suaeda salsa and was identified as Klebsiella aerogenes S27. Complete decolorization of MG (100 mg/l) was achieved in 8 h at 30 °C and pH 7.0. Ultraviolet-visible spectroscopy and Fourier-transform infrared spectroscopy analyses indicated the degradation of MG by the isolate. The enzymic assays of the strain showed the triphenylmethane reductase (TMR) activity. A gene encoding putative TMR-like protein (named as KaTMR) was cloned and heterologously expressed in Escherichia coli. KaTMR showed only 42.6-43.3% identities in amino acids compared with well-studied TMRs, and it phylogenetically formed a new branch in the family of TMRs. The degraded metabolites by recombinant KaTMR were detected by liquid chromatography-mass spectrometry, showing differences from the products of reported TMRs. The biotransformation pathway of MG was proposed. Phytotoxicity studies revealed the less-toxic nature of the degraded metabolites compared to the dye. This study presented the first report of an endophyte on the degradation and detoxification of triphenylmethane dye via a novel oxidoreductase, thus facilitating the study of the plant-endophyte symbiosis in the bioremediation processes.

摘要

内生微生物可以代谢有机污染物并促进植物生长,从而有助于受污染环境的植物修复。从湿地植物盐地碱蓬的叶片中分离到一种能够使孔雀石绿(MG)脱色的内生细菌,并鉴定为产气克雷伯氏菌 S27。该菌在 30°C 和 pH7.0 条件下 8 小时即可完全脱色 100mg/L 的 MG。紫外-可见光谱和傅里叶变换红外光谱分析表明该分离物对 MG 的降解作用。该菌株的酶促分析显示三苯甲烷还原酶(TMR)活性。克隆并在大肠杆菌中异源表达了一个编码假定 TMR 样蛋白的基因(命名为 KaTMR)。与研究充分的 TMR 相比,KaTMR 的氨基酸序列相似度仅为 42.6-43.3%,在 TMR 家族中形成了一个新的分支。通过液相色谱-质谱检测到重组 KaTMR 降解的代谢产物,与报道的 TMR 产物不同。提出了 MG 的生物转化途径。植物毒性研究表明,与染料相比,降解产物的毒性较低。本研究首次报道了一种内生菌通过一种新型氧化还原酶对三苯甲烷染料的降解和解毒作用,从而促进了植物-内生菌共生在生物修复过程中的研究。

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