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Exploring the application of Bacillus Subtilis Spore Surface Display System in the Field of Catechol Degradation
- 1 Shanghai Pinghe Bilingual School, Huangyang Road, Shanghai, China
* Author to whom correspondence should be addressed.
Abstract
Catechol is a pollutant commonly found in industrial and everyday wastewater, with genotoxic effects that can lead to mutations, DNA fragmentation, and chromosomal aberrations. Traditional physical and chemical degradation methods face challenges such as high costs and secondary pollution, whereas biological degradation offers a cost-effective and environmentally friendly alternative. This study investigates the use of the Bacillus subtilis spore surface display system for catechol degradation, aiming to develop an efficient, low-cost biodegradation method. Recombinant Bacillus subtilis was successfully engineered to display catechol 1,2-dioxygenase (C12O) and catechol 2,3-dioxygenase (C23O) on the spore surface. Immunoblotting and immunofluorescence confirmed protein display and enzyme activity assays demonstrated catechol degradation activity. The displayed enzymes showed stability across a wide pH and temperature range, maintaining activity even under methanol conditions. Compared to purified proteins, the spore-bound enzymes eliminated purification steps, reducing costs and improving degradation efficiency in some conditions. These results suggest that the Bacillus subtilis spore display system has great potential in biocatalysis, offering a novel strategy for environmental pollution remediation.
Keywords
Catechol degradation, spore surface display, catechol dioxygenase
[1]. Silva, A.S., et al., Enzymatic activity of catechol 1, 2-dioxygenase and catechol 2, 3-dioxygenase produced by Gordonia polyisoprenivorans. Quimica Nova, 2012. 35: p. 1587-1592.
[2]. Aravind, M.K., et al., Bioengineered Graphene Oxide Microcomposites Containing Metabolically Versatile Paracoccus sp. MKU1 for Enhanced Catechol Degradation. ACS Omega, 2020. 5(27): p. 16752-16761.
[3]. Topping, D.C., et al., Hydroquinone: acute and subchronic toxicity studies with emphasis on neurobehavioral and nephrotoxic effects. Food and chemical toxicology, 2007. 45(1): p. 70-78.
[4]. Zhou, Y., et al., Mesoporous carbon nitride based biosensor for highly sensitive and selective analysis of phenol and catechol in compost bioremediation. Biosensors and Bioelectronics, 2014. 61: p. 519-525.
[5]. Kubesch, K., et al., Degradation of catechol by ionizing radiation, ozone and the combined process ozone-electron-beam. Radiation Physics and Chemistry, 2005. 72(4): p. 447-453.
[6]. Ledesma, E.B., et al., An experimental study on the thermal decomposition of catechol. Proceedings of the Combustion Institute, 2002. 29(2): p. 2299-2306.
[7]. Xiao, J., et al., Enhancement of Fenton degradation by catechol in a wide initial pH range. Separation and Purification Technology, 2016. 169: p. 202-209.
[8]. Zhang, H., X. Bo, and L. Guo, Electrochemical preparation of porous graphene and its electrochemical application in the simultaneous determination of hydroquinone, catechol, and resorcinol. Sensors and Actuators B: Chemical, 2015. 220: p. 919-926.
[9]. El Azhari, N., et al., Molecular analysis of the catechol-degrading bacterial community in a coal wasteland heavily contaminated with PAHs. J Hazard Mater, 2010. 177(1-3): p. 593-601.
[10]. Vesely, M., et al., Analysis of catRABC operon for catechol degradation from phenol-degrading Rhodococcus erythropolis. Appl Microbiol Biotechnol, 2007. 76(1): p. 159-68.
[11]. Guo, G., et al., Isolation and characterization of two novel halotolerant Catechol 2, 3-dioxygenases from a halophilic bacterial consortium. Sci Rep, 2015. 5: p. 17603.
[12]. Wojcieszyńska, D., et al., Properties of catechol 2,3-dioxygenase from crude extract of Stenotrophomonas maltophilia strain KB2 immobilized in calcium alginate hydrogels. Biochemical Engineering Journal, 2012. 66: p. 1-7.
[13]. Bartels, J., et al., Sporobeads: The Utilization of the Bacillus subtilis Endospore Crust as a Protein Display Platform. ACS Synth Biol, 2018. 7(2): p. 452-461.
[14]. Wang, H., Y. Wang, and R. Yang, Recent progress in Bacillus subtilis spore-surface display: concept, progress, and future. Appl Microbiol Biotechnol, 2017. 101(3): p. 933-949.
[15]. Tavassoli, S., et al., Investigation of spore coat display of Bacillus subtilis beta-galactosidase for developing of whole cell biocatalyst. Arch Microbiol, 2013. 195(3): p. 197-202.
[16]. Wang, C.-L., S.-L. You, and S.-L. Wang, Purification and characterization of a novel catechol 1,2-dioxygenase from Pseudomonas aeruginosa with benzoic acid as a carbon source. Process Biochemistry, 2006. 41(7): p. 1594-1601.
[17]. Gou, M., et al., Characterization of catechol 1,2-dioxygenase from cell extracts of Sphingomonas xenophaga QYY. Science in China Series B: Chemistry, 2009. 52(5): p. 615-620.
[18]. Lin, J. and R.N. Milase, Purification and Characterization of Catechol 1,2-Dioxygenase from Acinetobacter sp. Y64 Strain and Escherichia coli Transformants. Protein J, 2015. 34(6): p. 421-33.
[19]. Han, L., S. Chen, and J. Zhou, Expression and cloning of catA encoding a catechol 1,2-dioxygenase from the 2,4-D-degrading strain Cupriavidus campinensis BJ71. Prep Biochem Biotechnol, 2020. 50(5): p. 486-493.
[20]. Li, J., et al., Expression and characterization of catechol 1,2-dioxygenase from Oceanimonas marisflavi 102-Na3. Protein Expr Purif, 2021. 188: p. 105964.
[21]. Guzik, U., et al., High activity catechol 1,2-dioxygenase from Stenotrophomonas maltophilia strain KB2 as a useful tool in cis,cis-muconic acid production. Antonie Van Leeuwenhoek, 2013. 103(6): p. 1297-307.
[22]. Adewale, P., et al., A novel Bacillus ligniniphilus catechol 2,3-dioxygenase shows unique substrate preference and metal requirement. Sci Rep, 2021. 11(1): p. 23982.
Cite this article
Fu,Z. (2025). Exploring the application of Bacillus Subtilis Spore Surface Display System in the Field of Catechol Degradation. Theoretical and Natural Science,99,1-15.
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Volume title: Proceedings of the 5th International Conference on Biological Engineering and Medical Science
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