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Heterologous surface display of Serratia marcescens-derived laccase and its decolorization performance toward Azo
Aims:
Laccase represents a high-value green catalyst with extensive industrial application prospects. The aim of this study is to construct a laccase surface-displaying recombinant strain LacA-BL21 by fusing LacA from Serratia marcescens with Escherichia coli (E. coli) Ag43 autotransport system, optimize its enzyme production and dye decolorization parameters, verify the surface display system efficiency, and evaluate its potential for industrial wastewater bioremediation.
Methods And Results:
In this study, we generated the surface-displaying strain LacA-BL21 by fusing laccase (LacA) from S. marcescens with the Ag43 autotransport system from E. coli. The enzyme activity results indicate that the recombinant laccase exhibits optimal catalytic activity at 45°C and pH 4.0. Key parameters affecting enzyme production were systematically optimized and LacA-BL21 achieved a maximum laccase activity of 43.66 ± 1.45 U ml-1 in TB medium supplemented with 4% methanol, 1.77 times that of the wild-type strain ZH-5. Furthermore, qRT-PCR (quantitative real-time polymerase chain reaction) analysis demonstrated that the relative transcription level of the target gene in the LacA-BL21 was 1.91-fold higher than that in the wild-type strain ZH-5, verifying the high efficiency of the surface display system for laccase heterologous expression. Dye decolorization assays using simulated wastewater were performed to evaluate the catalytic performance of the recombinant LacA-BL21 system, and the results indicated outstanding decolorization efficacy toward various dye substrates. Following systematic process optimization, the crude preparation containing recombinant LacA achieved an 82.17% decolorization rate for Congo red simulated wastewater within 2 h.
Conclusion:
This study constructs a surface display system for laccase expression, which exhibits exceptional catalytic efficiency and favorable decolorization capability toward azo dyes, thereby harboring substantial promise for the bioremediation of industrial wastewater.
