Optimization of Solid State Fermentation of 2-Deoxy-D-Glucose Aspergillus niger Mutant for Improved Cellulase Production
Abstract
Mutation and optimization of solid state fermentation of Aspergillus niger were performed in order to enhance the production of lignocellulolytic enzymes for biotransformation of cellulosic wastes. Spores of the fungus were irradiated with ultraviolet rays at 300μW/cm2 at a distance of 15cm for 30 minutes. Mutants were selected in mineral salt medium containing 2-deoxy D-glucose (0.001 g/L). The selected A. niger mutant strain had a 2.1-fold increase in cellulase production more than the wild type. Solid-state fermentation of the selected strain was carried out and the optimal conditions for the production of cellulase were investigated. Optimum moisture for high enzyme yield was 20 %, temperature of 37ºC, and a pH of 3.5. Other physical parameters included fermentation time of 4 days, inoculum size of 25% w/w and age of seeding culture of 4 days. When the medium was supplemented with corn-starch, glucose, lactose, sucrose and maltose; corn starch [9.21 U/mL and 2.30 U/mL Carboxylmethyl cellulose (CMCase) and total cellulose (FPase) respectively] enhanced the production of cellulase more than other substrates. Surfactants used were SDS, EDTA, Tween-20 and Tween-80. EDTA (9.39 U/mL and 2.30 U/mL CMCase and FPase respectively) enhanced enzyme production more than other surfactants. When ZnSO4, FeSO4, CuSO4, CaCl2, MgCl2, CoCl2 and KCl were used; FeSO4 (9.29 U/mL and 2.46 U/mL CMCase and FPase respectively) supported cellulase production over others. Urea (9.52 U/mL and 2.37 U/mL CMCase and FPase respectively) was the best nitrogen supplement compared to malt extract, yeast extract, NH4NO3 and NH4Cl.
Downloads
References
Abu, E. A., Ameh, D. A., Onyenekwe, P. C., Agbaji, A. S., Ado, S. A. and Nwaeze, A. R. (2002). Comparative studies of the effect of pretreatments of three Agrowastes on cellulose production by Aspergillus niger SL1. Scientia Africana 15(1): 57-63.
Archaya, P. B., Achaya, D. K. and Modi, H. A. (2008). Optimiztion or cellulase production by Aspergillus niger using saw dust as substrate. African Journal of Biotechnology 7(22): 4147-4152.
Anwar, M. N., Suto, M. and Tomida, F. (1996). Isolation of mutants of Penicillium urpurogenum resistant to catabolite repression. Applied Microbiology and Biotechnology 45: 684-687.
Ballesteros, I., Oliva, J. M., Carrasco, J., Cabanas, A., Navarro, A. A. and Ballesteros, M. (1998). Effect of surfactants and zeolites on simultaneous saccharification and fermentation of steam-exploded poplar biomass to ethanol. Applied Biochemistry and Biotechnology 70: 369-381.
De Nicolas-Santiago, S., Regalado-Gonzalez, C., Garcia-Almendarez, B., Fernandez, F. J., Tellez-Jurado, A. and Huerta-Ochoa, S. A. (2006). Physiological, morphological, and mannanase production studies on Aspergillus niger uam-gs 1 mutants. Electronic Journal of Biotechnology 9(1): 50-60.
Dillion, A. J. P., Zorgi, C., Camassola, M., and Henriques, J. A. P. (2005). Use of 2-deoxyglucose in liquid media for the selection of mutant strains of Penicillium echinulatum producing increased cellulase and β-glucosidase activities. Applied Microbiology and Biotechnology 70:740-746.
Farkas, V., Labudova, I., Bauers, S. and Ferenczy, L. (1981). Preparation of mutants of Trichoderma viride with increased production of cellulase. Folia Microbiology 26:105-110.
Howard, R. L., Abotsi, E., Jansen Van Rensburg, E. L. and Howard, S. (2003). Lignocellulosic Biotechnology: issues of bioconveion and enzyme production. African Journal of Biotechnology 2(12): 602-619.
Junior, M. M., Batistote, M., Cilli, E. M. and Ernandes, J. R. (2009). Sucrose fermentation by Brazillian ethanol production yeast in media containing structural complex nitrogen sources. Journal of the Institute of brewing 115: 191-197.
Kaur, J., Chandha, B. S., Kumar, B. A. Kaur, G. S. and Saini, H. S. (2007). Purification and characterization of β-glucosidase from Melanocarpus Sp. MTCC3922. Electronic Journal of Biotechnology 10: 260-270.
Mehdi, D., Heidi, S. and Wensheng, Q. (2009). Fungal bioconversion of lignocellulosic residues; Opporunities and Perspectives. International Journal of Biological Sciences 5(6): 578-595.
Montiel-Gonzalez, A. M., Viniegra-Gonzalez, G, Fernandez, F. J. and Leora, O. (2004). Effect of water activity on invertase production in solid state fermentation by improved diploid strains of Aspergillus niger. Process Biochemistry 39(12): 2085-2090.
Narasimha, G., Sridevi, A., Viswanath, B., Chandra, M. S. and Reddy, R. B. (2006). Nutrient effects on production of cellulolytic enzymes by Aspergillus niger. African Journal of Biotechnology 5(5): 472-476
Nikolic, S., Mojovic, L., Pejin, D., Rakin, M. and Vucurovic, V. (2009). Improvement of ethanol fermentation of corn Semolina hydrolyzates with immobilized yeast by medium supplementation. Food Technology and Biotechnology 47: 83-89.
Pandey, A., Selvakumar, P., Soccol, C. R., and Nigam, P. (1999). Solid state fermentation for the production of industrial enzymes. Current Science 77: 149-162.
Parekh, S., Vinci, V. A. and Strobel, R. J. (2000). Improvement of microbial strains and fermentation processes. Applied Microbiology and Biotechnology 54(3): 287-301.
Pradeep, M. R., Janardhan, A., Kumar, A. P. and Narasimha, G. (2012). Induction of chemical mutations in Aspergillus niger to enhance cellulase production. International Journal of Environmental Biology 2(3): 129-132.
Singh, A., Singh, N. and Bishnoi, N. R. (2009). Production of cellulases by Aspergillus heteromorphous from wheat straw under submerged fermentation. International Journal of Environmental Science and Engineering 1(1):23-26.
Sun Y. and Cheng, J. (2002). Hydrolysis of lignocellulosic material from ethanol production: A review. Bioresource Technology 83: 1-11.
Vu, V. H., Pham, T. A. and Kim, K. (2009). Fungal strain improvement for cellulase production using repeated and sequential mutagenesis. Mycobiology 37(4): 267-271.
Vu, V. H., Pham, T. A. and Kim, K. (2010a). Improvement of fungal cellulase production by mutation and optimization of Solid State Fermentation. Mycobiology 39(1): 20-25.
Vu, V. H., Pham, T. A. and Kim, K. (2010b) Improvement of fungal strain by repeated and sequential mutagenesis and optimization of solid-state fermentation for the hyper-production of raw starch digesting enzyme. Journal of Microbiology and Biotechnology 20: 718-726.
Yang, B. and Wyman, C. E. (2006). BSA treatment to enhance enzymatic hydrolysis of cellulose in lignin containing substrates Biotechnology and Bioengineering 94(4):611-617.
Zhu, H., Zhang, H., Zhang, Y. and Huang F. (2011). Lignocellulose degradation, enzyme production and protein enrichment by Trametes versicolor during solid-state fermentation of corn stover. African Journal of Biotechnology 10(45): 9182-9192.
Additional Files
Published
Issue
Section
Categories
License
Copyright (c) 2023 Babalola, O. M., Magbagbeola, O. A., Ebuehi, O. A. T., Okochi, V. I., Okafor, U. A.
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
