Response Surface Based Optimization of Laccase Production from Cerena Unicolor with Wheat Straw as a Substrate
Keywords:
RSM, CCD, OFAT, optimization, laccase, ABTSAbstract
Laccase currently has a global market of more than 3 million USD, due to its redox potential, whose cost is determined by the raw material. Fungal laccase was produced from cheap agrowaste using a fungal source. Fungal isolation was done from microbes of decomposing cellulosic raw material. Cerrena unicolorwas found to be the most promising fungus and identified using ITS based sequencing, phylogenetic analysis and morphological study. Among different cellulosic materials, wheat straw was a better raw material for fungal isolates in laccase production. The optimization of laccase production was done using wheat straw with one factor at a time to find optimaltemperature (30 °C), incubation period (14 days), moisture percent (80) and pH (5). Response surface method with central composite design using yeast extract as an organic source of nitrogen or ammonium sulphate as an inorganic source was compared for laccase production. Copper sulphate was used as an inducer. It gave a significant model with quadratic equation which gave nearly similar experimental and predicted laccase production values. A 6-fold change in laccase production was found after optimization. The purification product after SDS PAGE showed the isolated laccase to have a single polypeptide of around 64 kDa. The study found that copper acted as an inducer in the laccase production, where solid state fermentation using wheat straw was a compelling combination for the isolated fungi in laccase production. The isolated fungi have the potential to produce laccase as a result of different growth parameters and the use of different agricultural wastes.
References
Adekunle, A., Guo, C., & Liu, C.Z. (2017). Lignin-Enhanced Laccase Production from Trametes versicolor. Waste and Biomass Valorization, 8. https://doi.org/10.1007/s12649-016-9680-4
Adekunle, A., Zhang, C., Guo, C., & Liu, C.Z. (2017). Laccase Production from Trametes versicolor in Solid-State Fermentation of Steam-Exploded Pretreated Cornstalk. Waste and Biomass Valorization, 8. https://doi.org/10.1007/s12649-016-9562-9
Alcalde, M., & Bulter, T. (2003). Colorimetric assays for screening laccases. Directed Enzyme Evolution: Screening and Selection Methods, 193-201. https://doi.org/10.1385/1-59259-396-8:193
Arregui, L. et al. Laccases: structure, function, and potential application in water bioremediation. Microb Cell Fact 18, 200 (2019). https://doi.org/10.1186/s12934-019-1248-0
Aruna, S., Gomathi, M., & Gehitha, T. R. (2012). Screening, production and partial purification of laccase enzyme from white rot fungi. Indian Journal of Current research, 1 (2), 10-17.
Chauhan, P. S., Goradia, B., & Jha, B. (2018). Optimization and up scaling of ionic liquid tolerant and thermo-alkali stable laccase from a marine Staphylococcus arlettae S1-20 using tea waste. Journal of the Taiwan Institute of Chemical Engineers, 86, 1-8. https://doi.org/10.1016/j.jtice.2018.02.032
Chauhan, P. S., & Jha, B. (2018). Pilot scale production of extracellular thermo‐alkali stable laccase from Pseudomonas sp. S2 using agro waste and its application in organophosphorous pesticides degradation. Journal of Chemical Technology & Biotechnology, 93(4), 1022-1030. https://doi.org/10.1002/jctb.5454
Eichlerová, I., Šnajdr, J., & Baldrian, P. (2012). Laccase activity in soils: considerations for the measurement of enzyme activity. Chemosphere, 88(10), 1154-1160.
Faulina, S. A. et al. (2020). The higher laccase enzyme producer, Cerrena sp. BMd. TA. 1, isolated from Gunung Rinjani National Park, West Nusa Tenggara, Indonesia. Biodiversitas Journal of Biological Diversity, 21(8). https://doi.org/10.13057/biodiv/d210853
Gallagher, S. R. (2006). One‐dimensional SDS gel electrophoresis of proteins. Current protocols in immunology, 75(1), 8.4. 1-8.4. 37. https://doi.org/10.1002/0471142735.im0804s75
Ghosh, P., & Ghosh, U. (2017). Statistical optimization of laccase production by Aspergillus flavus PUF5 through submerged fermentation using agro-waste as cheap substrate. Acta Biologica Szegediensis, 61(1), 25-33.
Ire, F. S., & Ahuekwe, E. F. (2016). Production of fungal laccase using orange peelings as substrate by submerged static fermentation. British Microbiology Research Journal, 15(5), 1-19.
Janusz, G. et al. (2012). Cloning and characterization of a laccase gene from biotechnologically important basidiomycete Cerrena unicolor. https://doi.org/10.5109/22046
Kiiskinen, L. L., Rättö, M., & Kruus, K. (2004). Screening for novel laccase‐producing microbes. Journal of applied microbiology, 97(3), 640-646. https://doi.org/10.1111/j.1365-2672.2004.02348.x
Kolomytseva, M. et al. (2017). Rapid identification of fungal laccases/oxidases with different pH-optimum. Process Biochemistry, 62, 174-183. https://doi.org/10.1016/j.procbio.2017.07.027
Kumar, S. et al. (2018). MEGA X: molecular evolutionary genetics analysis across computing platforms. Molecular biology and evolution, 35(6), 1547. https://doi.org/ 10.1093/molbev/msy096
Liu, Z. et al. (2009). A newly isolated Paecilomyces sp. WSH-L07 for laccase production: isolation, identification, and production enhancement by complex inducement. Journal of Industrial Microbiology and Biotechnology, 36(10), 1315-1321.
Mann, J. et al. (2015). Use of olive mill wastewater as a suitable substrate for the production of laccase by Cerrena consors. International Biodeterioration & Biodegradation, 99, 138-145.
Margot, J. et al. (2013). Bacterial versus fungal laccase: potential for micropollutant degradation. AMB Express, 3(1), 63. https://doi.org/10.1186/2191-0855-3-63
Mishra, V. et al. (2017). Enhancement in multiple lignolytic enzymes production for optimized lignin degradation and selectivity in fungal pretreatment of sweet sorghum bagasse. Bioresour Technol, 236, 49-59. https://doi.org/10.1016/j.biortech.2017.03.148
Muthukumarasamy, N. P., & Murugan, S. (2014). Production, purification and application of bacterial laccase: a review. Biotechnology, 13(5), 196. https://doi.org/ 10.3923/biotech.2014.196.205
Nambisan, P. (2018). Data of optimization of laccase production by Marasmiellus palmivorus LA1 under solid state fermentation using one factor at a time method. Data in brief, 17, 1276-1282. https://doi.org/10.1016/j.dib.2018.02.011
Patel, H., & Gupte, A. (2016). Optimization of different culture conditions for enhanced laccase production and its purification from Tricholoma giganteum AGHP. Bioresources and Bioprocessing, 3, 1-10.
Rivera-Hoyos, C. M., Morales-Álvarez, E. D., Poutou-Piñales, R. A., Pedroza-Rodríguez, A. M., RodrÍguez-Vázquez, R., & Delgado-Boada, J. M. (2013). Fungal laccases. Fungal Biology Reviews, 27(3-4), 67-82.
Sharma, S., & Murty, D. S. (2021). Enhancement of Laccase Production by Optimizing the Cultural Conditions for Pleurotus sajor-caju in Solid-State Fermentation. Journal of Pure & Applied Microbiology, 15(2).https://doi.org/10.22207/JPAM.15.2.54
Tekpinar, A. D., & Kalmer, A. (2019). Utility of various molecular markers in fungal identification and phylogeny. Nova Hedwigia, 109(1-2), 187-224.
Terrón, M. C. et al. (2004). Structural close-related aromatic compounds have different effects on laccase activity and on lcc gene expression in the ligninolytic fungus Trametes sp. I-62. Fungal Genetics and Biology, 41(10), 954-962. https://doi.org/10.1016/j.fgb.2004.07.002
Vijayaraghavan, P., & Prakash Vincent, S. G. (2014). Medium optimization for the production of fibrinolytic enzyme by Paenibacillus sp. IND8 using response surface methodology. The Scientific World Journal, 2014. https://doi.org/10.1155/2014/276942
Wang, F. et al. (2019). Fungal Laccase Production from Lignocellulosic Agricultural Wastes by Solid-State Fermentation: A Review. Microorganisms, 7(12). https://doi.org/10.3390/microorganisms7120665
Wang, Z. et al. (2016). A comparison of chemical treatment methods for the preparation of rice husk cellulosic fibers. International Journal of Environmental & Agriculture Research, 2(1), 2454-1850. https://doi.org/10.1155/2014/276942
Wood, D. (1980). Production, purification and properties of extracellular laccase of Agaricus bisporus. Microbiology, 117(2), 327-338. https://doi.org/10.1099/00221287-117-2-327
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