Surface Display of Bacterial Laccase CotA on Escherichia coli Cells and its Application in Industrial Dye Decolorization
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Laccase CotA from Bacillus subtilis 168 was successfully displayed on the membrane of Escherichia coli cells using poly-γ-glutamate synthetase A protein (PgsA) from B. subtilis as an anchoring matrix. Further analyses demonstrated that the fusion protein PgsA/CotA efficiently translocates to the cell surface of E. coli with an enzymatic activity of 65 U/108 cells. Surface-displayed CotA was shown to possess improved enzymatic properties compared with those of the wild-type CotA, including higher thermal stability (above 90% activity at 70 °C and nearly 40% activity at 90 °C after 5-h incubation) and stronger inhibitor tolerance (approximately 80 and 65% activity when incubated with 200 and 400 mM NaCl, respectively). Furthermore, the whole-cell system was demonstrated to have high enzymatic activity against anthraquinone dye, Acid Blue 62, triphenylmethane dye, Malachite Green, and azo dye, Methyl Orange with the decolorization percentages of 91, 45, and 75%, after 5-h incubation, respectively.
KeywordsCell-surface display Bacterial laccase CotA Synthetic dye Decolorization
This work was supported by the National Natural Science Foundation of China (No. 31700092, No. 21727818, No. 21390200, No. 21706125, No. 21706124), the Jiangsu Province Natural Science Foundation for Youths (No. BK20170997, No. BK20170993), the Project of State Key Laboratory of Materials Oriented Chemical Engineering (KL17-09) and the Jiangsu Synergetic Innovation Center for Advanced Bio-Manufacture (XTE1834), the open foundation of Jiangsu Key Laboratory for Biomass-Based Energy and Enzyme Technology, (BEETKB1801).
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