The ABC Transporter Protein from Bacillus safensis Improves the Manganese Tolerance and Biofilm Formation in Escherichia coli
LIU Cheng-Jie1, NIU Xi1, HUANG Shi-Hui2, WANG Jia-Fu1,*, RAN Xue-Qin2
1 College of Life Sciences/Institute of Agricultural Bioengineering/Key Laboratory of Conservation and Innovation of Highland Mountain Plant Resources of Ministry of Education, Guizhou University, Guiyang 550025, China; 2 College of Animal Science, Guizhou University, Guiyang 550025, China
Abstract：The ATP-binding cassette transporter protein (ABCT) in cells plays a crucial role in the tolerance of toxic substances, which participates in bacterial growth and the elimination of toxic compounds. Bacillus safensis ST7 has strong manganese oxidation ability, and the ABCT (Gene ID: 46572106) was significantly upregulated under manganese stress, which may be related to the manganese tolerance of Bacillus safensis. In order to analyze the role of ABCT in manganese stress in B. safensis, PCR, qPCR, and SDS-PAGE methods were used to clone the ABCT gene in B. safensis ST7. Overexpression vectors were constructed and transformed into Escherichia coli BL21(DE3). The growth curve, biofilm formation ability, manganese tolerance, and manganese oxidation activity of the recombinant strain were measured to investigate the effect of heterologous expression of the ABCT gene on the manganese tolerance and manganese oxidation activity of E. coli. The results showed that the recombinant strain carried the complete ABCT gene, and the expression level of the recombinant plasmid pET28a-ABCT was significantly upregulated in BL21(DE3) cells (P<0.05). Its growth was not affected, and its biofilm formation ability was enhanced. Under manganese stress, the expression of ABCT gene increased by 1.41 times, with time and concentration dependence. Compared with the control strain, the recombinant strain showed an increase of over 1.5 times in manganese tolerance concentration and manganese oxidation activity. The results suggested that the ABCT gene of B. safensis ST7 was involved in the manganese transport and biofilm formation process in E. coli. Overexpression of this gene could enhance the oxidative activity and tolerance of E. coli cells to manganese (Ⅱ). This study is important for the development and application of manganese-oxidizing bacteria.
刘成洁, 牛熙, 黄世会, 王嘉福, 冉雪琴. 沙福芽胞杆菌ABC转运蛋白促进大肠杆菌的锰耐受和生物膜形成[J]. 农业生物技术学报, 2025, 33(5): 1130-1140.
LIU Cheng-Jie, NIU Xi, HUANG Shi-Hui, WANG Jia-Fu, RAN Xue-Qin. The ABC Transporter Protein from Bacillus safensis Improves the Manganese Tolerance and Biofilm Formation in Escherichia coli. 农业生物技术学报, 2025, 33(5): 1130-1140.
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