Wang Lijuan, Peng Rihe, Tian Yongsheng, Xu Jing, Wang Bo, Han Hongjuan, Fu Xiaoyan, Gao Jianjie, Yao Quanhong
Shanghai Key Laboratory of Agricultural Genetics and Breeding, Biotechnology Research Institute of Shanghai Academy of Agricultural Sciences, 2901 Beidi Road, Shanghai, China.
AMB Express. 2022 May 14;12(1):55. doi: 10.1186/s13568-022-01396-9.
As a kind of refractory organic pollutant, 4-fluorophenol (4-FP) can be degraded by only a few microorganisms with low efficiency because of the great electron-withdrawing ability of fluorine atoms. So it is necessary to artificially construct engineered strain to improve the degradation efficiency and meet the requirements of pollutant degradation. In this study, four genes (fpdA2, fpdB, fpdC, and fpdD) for 4-FP degradation from Arthrobacter sp. strain IF1 were optimized and synthesized and then reconstructed into Escherichia coli by a multi-monocistronic vector to obtain recombinant BL-fpd that could degrade 4-FP efficiently. Under optimized induction conditions (inducing the strain by 2 g/L L-arabinose and 1 mM IPTG at 37 ℃), BL-fpd could completely degrade 2 mM 4-FP, 4-chlorophenol, 4-bromophenol, and 4-nitrophenol into β-ketoadipate, which could be further metabolized by the bacteria. FpdA2 showed the highest activity towards 4-bromophenol. The strain could completely degrade 1 mM 4-FP in industrial wastewater within 3 h. This study provided a promising strain for the degradation of 4-FP and some other 4-substituted phenols. The construction technologies of multi-monocistronic expression vector may also be used to construct other organic pollutants degrading bacteria.
作为一种难降解的有机污染物,4-氟苯酚(4-FP)由于氟原子具有很强的吸电子能力,只能被少数微生物降解且效率较低。因此,有必要人工构建工程菌株以提高降解效率,满足污染物降解的需求。在本研究中,对节杆菌属菌株IF1中4-FP降解的四个基因(fpdA2、fpdB、fpdC和fpdD)进行了优化合成,然后通过多顺反子载体将其重组到大肠杆菌中,获得了能够高效降解4-FP的重组菌株BL-fpd。在优化的诱导条件下(在37℃用2g/L L-阿拉伯糖和1mM IPTG诱导菌株),BL-fpd能够将2mM的4-FP、4-氯苯酚、4-溴苯酚和4-硝基苯酚完全降解为β-酮己二酸,β-酮己二酸可被细菌进一步代谢。FpdA2对4-溴苯酚表现出最高活性。该菌株能够在3小时内完全降解工业废水中1mM的4-FP。本研究为4-FP和其他一些4-取代酚的降解提供了一种有前景的菌株。多顺反子表达载体的构建技术也可用于构建其他有机污染物降解菌。