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在沼泽红假单胞菌CGA009上进行CadR蛋白的表面展示以增强重金属生物修复能力。

Surface display of CadR protein on Rhodopseudomonas palustris CGA009 for enhanced heavy metal bioremediation.

作者信息

Wang Minmin, Gao Zixuan, Jiang Qiushi, Mu Xuefang, Jiang Youmin, Li Yanjing, Zhang Sihu, Cao Wen, Guo Liejin

机构信息

State Key Laboratory of Multiphase Flow in Power Engineering, Xi'an Jiaotong University, Xianning West Road, Xi'an 710049, China.

State Key Laboratory of Multiphase Flow in Power Engineering, Xi'an Jiaotong University, Xianning West Road, Xi'an 710049, China.

出版信息

J Hazard Mater. 2025 Aug 15;494:138592. doi: 10.1016/j.jhazmat.2025.138592. Epub 2025 May 12.

Abstract

Heavy metal pollution poses significant ecological and public health risks, and surface display engineering shows promise for bioremediation in this area. Although anoxygenic photosynthetic purple nonsulfur bacteria (PNSB) have been effectively applied to degrade pollutants due to their metabolic versatility, the use of surface display technology in PNSB remains very limited. In this study, we constructed a surface display system using Rhodopseudomonas palustris CGA009 as the host. The metal-binding protein CadR was fused with outer membrane protein A (OmpA) and expressed in CGA009. SDS-PAGE and immunofluorescence analysis identified the successful expression of the fusion protein on the cell surface. In addition, we used flow cytometry to explore the enhancement effects of linker peptides and different promoters on surface display efficiency under different light intensities. The surface display system enhanced the heavy metal resistance of the host bacteria, and the maximum removal rate of Cd reached 95.6 %. By means of Langmuir isotherm analysis, the maximum biosorption capacity of the system for Cd is 101.11 mg/g. The system demonstrates feasibility for application in complex real-world environmental samples. The presence of various metal ions does not interfere with the system's specific adsorption of Cd²⁺. It can stably maintain an adsorption efficiency of over 80 % under conditions of pH 6-8, temperatures of 20-35°C, and light intensity of 1000-6000 lux. Additionally, the system achieves a removal efficiency of 94.3 % in Cd wastewater. In summary, this study provides a reference for the development of photosynthetic bacterial surface display systems and provides an advanced bioremediation strategy for heavy metal contaminated wastewater.

摘要

重金属污染带来了重大的生态和公共健康风险,而表面展示工程在该领域的生物修复方面显示出了潜力。尽管无氧光合紫色非硫细菌(PNSB)因其代谢的多功能性已被有效地应用于污染物降解,但表面展示技术在PNSB中的应用仍然非常有限。在本研究中,我们构建了一个以沼泽红假单胞菌CGA009为宿主的表面展示系统。将金属结合蛋白CadR与外膜蛋白A(OmpA)融合,并在CGA009中表达。SDS-PAGE和免疫荧光分析确定了融合蛋白在细胞表面的成功表达。此外,我们使用流式细胞术探索了连接肽和不同启动子在不同光照强度下对表面展示效率的增强作用。该表面展示系统增强了宿主细菌对重金属的抗性,Cd的最大去除率达到95.6%。通过朗缪尔等温线分析,该系统对Cd的最大生物吸附容量为101.11 mg/g。该系统证明了在复杂的实际环境样品中应用的可行性。各种金属离子的存在不会干扰该系统对Cd²⁺的特异性吸附。在pH值为6-8、温度为20-35°C、光照强度为1000-6000勒克斯的条件下,它可以稳定地保持超过80%的吸附效率。此外,该系统在Cd废水中的去除效率达到94.3%。总之,本研究为光合细菌表面展示系统的开发提供了参考,并为重金属污染废水提供了一种先进的生物修复策略。

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