Development of a Novel Micro-Aerobic Cultivation Strategy for High Potential CotA Laccase Production
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Recently, bacterial laccases has drawn researchers’ interest due to their ability to overcome high pH and salt concentration conditions compared to fungal laccases. Here we report a novel micro-aerobic cultivation strategy for enhancing CotA laccase expression and study its application for dye decolorization. Micro-aerobic cultivation of Escherichia coli BL21 (DE3) strain carrying pT7-FLAG-MAT-TAG-1-CotA had significantly enhanced CotA laccase activity up to 13903 U/L. The most unique findings of this investigation are that micro-aerobic cultivation strategy enhanced the reactive oxygen species production which consequently led to the over expression of CotA laccase gene. Malachite green, Crystal violet, Congo red and Bromophenol blue were efficiently decolorized by using purified CotA laccase without presence of any mediators at pH 6 and 9. These results provide a great platform for the dynamic production and application of bacterial laccase in industry.
KeywordsBacillus subtilis Bacterial laccase CotA IPTG Micro-aerobic condition ROS
The funding of this study was supported by the National Key Technology Research and Development Program of China (No. 2015BAK45B01), the National Natural Science Foundation of China (No. 21476242), and the National Basic Research Program (973 Program) of China (No. 2013CB733600). We would like to acknowledge the Chinese Government for the financial support of the Ph. D scholarship awarded to Nadia A. Samak.
- 12.Durão, P., Chen, Z., Fernandes, A.T., Hildebrandt, P., Murgida, D.H., Todorovic, S., Pereira, M.M., Melo, E.P., Martins, L.O.: Copper incorporation into recombinant CotA laccase from Bacillus subtilis: characterization of fully copper loaded enzymes. J. Biol. Inorg. Chem. 13, 183–193 (2008)CrossRefGoogle Scholar
- 14.Constantinidou, C., Hobman, J.L., Griffiths, L., Patel, M.D., Penn, C.W., Cole, J.A., Overton, T.W.: A reassessment of the FNR regulon and transcriptomic analysis of the effects of nitrate, nitrite, NarXL, and NarQP as Escherichia coli K12 adapts from aerobic to anaerobic growth. J. Biol. Chem. 281(8), 4802–4815 (2006)CrossRefGoogle Scholar
- 32.Artsatbanov, V., Vostroknutova, G., Shleeva, M., Goncharenko, A., Zinin, A., Ostrovsky, D., Kapreliants, A.: Influence of oxidative and nitrosative stress on accumulation of diphosphate intermediates of the non-mevalonate pathway of isoprenoid biosynthesis in corynebacteria and mycobacteria. Biochemistry 77, 362–371 (2012)Google Scholar
- 35.Xu, F.: Oxidation of phenols, anilines and benzenethiols by fungal laccases: correlation between activity and redox potentials as well as halide inhibition. Biochemistry 35, 7608–7614 (1996)Google Scholar
- 38.Liu, H., Cheng, Y., Du, B., Tong, C., Liang, S., Han, S., Zheng, S., Lin, Y.: Overexpression of a novel thermostable and chloride-tolerant laccase from Thermus thermophilus SG0.5JP17-16 in Pichia pastoris and its application in synthetic dye decolorization. PLoS ONE (2015). doi: 10.1371/journal.pone.0119833 CrossRefGoogle Scholar