Abstract
Properties of beta-glucosidase produced by Aspergillus niger URM 6642 recently isolated from the Atlantic rainforest biome and its potential tolerance to saccharification of lignocellulosic biomass products and fermentation inhibitors was evaluated. The fungus was cultivated under solid state culture conditions at 37°C with different agro-industrial wastes. High levels of beta-glucosidase (3778.9 U g−1)from A. niger were obtained with rice meal as substrate under solid state culture conditions after ten days. Optimum pH for this particular beta-glucosidase activity was 4.0 although it was stable in the range of 4.0 to 7.0. The half-life (T½) of beta-glucosidase at 55°C is 3 h. However, at the optimum temperature of the enzyme, 65°C, T½ is 20 min. The enzyme showed tolerance to various compounds such as glucose, xylose, 5-hydroxymethyl furfural, furfural, coumarin, ethanol and acetic acid. Therefore, beta-glucosidase from the novel A. niger species may be of potential use in the saccharification of lignocellulosic biomass, as well as an additional enzyme supplement in cellulase cocktails used to increase the yield of fermentable sugars.
Similar content being viewed by others
References
Gao, Z. Q., Van Hop, D., Yen, L. T., Ando, K., Hiyamuta, S., & Kondo, R. (2012). The production of β-glucosidases by Fusarium proliferatum NBRC109045 isolated from Vietnamese forest. AMB Express, 2, 49. DOI: 10.1186/2191-0855-2-49.
Gao, L., Gao, F., Zhang, D. Y., Zhang, C., Wu, G. H., & Chen, S. L. (2013). Purification and characterization of a new β-glucosidase from Penicillium piceum and its application in enzymatic degradation of delignified corn stover. Bioresource Technology, 147, 658–661. DOI: 10.1016/j.biortech.2013.08.089.
Harnicharnchai, P., Champreda, V., Sornlake, W., & Eurwilaichitr, L. (2009). A thermotolerant β-glucosidase isolated from an endophytic fungi, Periconia sp., with a possible use for biomass conversion to sugars. Protein Expression and Purification, 67, 61–69. DOI: 10.1016/j.pep.2008.05.022.
Jørgensen, H., Vibe-Pedersen, J., Larsen, J., & Felby, C. (2007). Liquefaction of lignocellulose at high-solids concentrations. Biotechnology and Bioengineering, 96, 862–870. DOI: 10.1002/bit.21115.
Kaushal, R., Sharma, N., & Tandon, D. (2012). Cellulase and xylanase production by co-culture of Aspergillus niger and Fusarium oxysporum utilizing forest waste. Turkish Journal of Biochemistry, 37, 35–41. DOI: 10.5505/tjb.2012.43434.
Kaur, J., Chadha, B. S., Kumar, B. A., Kaur, G. S., & Saini, H. S. (2007). Purification and characterization of β-glucosidase from Melanocarpus sp. MTCC 3922. Electronic Journal of Biotechnology, 10(2), 4. DOI: 10.2225/vol10-issue2-fulltext-4.
Liu, D. Y., Zhang, R. F., Yang, X. M., Zhang, Z. H., Song, S., Miao, S. Z., & Shen, Q. R. (2012). Characterization of a thermostable β-glucosidase from Aspergillus fumigatus Z5, and its functional expression in Pichia pastoris X33. Microbial Cell Factory, 17, 11–25. DOI: 10.1186/1475-2859-11-25.
Ma, S. J., Leng, B., Xu, X. Q., Zhu, X. Z., Shi, Y., Tao, Y. M., Chen, S. X., Long, M. N., & Chen, Q. X. (2011). Purification and characterization of β-1,4-glucosidase from Aspergillus glaucus. African Journal of Biotechnology, 10, 19607–19614. DOI: 10.5897/ajb11.2144.
Masui, D. C., Zimbardi, A. L. R. L., Souza, F. H. M., Guimarães, L. H. S., Furriel, R. P. M., & Jorge, J. A. (2012). Production of a xylose-stimulated β-glucosidase and a cellulase free thermostable xylanase by the thermophilic fungus Humicola brevis var. thermoidea under solid state fermentation. World Journal Microbiology & Biotechnology, 28, 2689–2701. DOI: 10.1007/s11274-012-1079-1.
Michelin, M. (2013). Aplication of lignocelulosic residues in the production of cellulase and hemicellulase from fungi. In T. Mahendra, & M. L. T. M. Polizelli (Eds.), Fungal enzymes (pp. 32–59). Boca Raton, FL, USA: Taylor & Francis.
Miller, G. L. (1959). Use of dinitrosalicilic acid reagent for determination of reducing sugar. Analytical Chemistry, 31, 426–428. DOI: 10.1021/ac60147a030.
Nascimento, C. V., Souza, F. H. M., Masui, D. C., Leone, F. A., Peralta, R. M., Jorge, J. A., & Furriel, R. P. M. (2010). Purification and biochemical properties of a glucose-stimulated β-d-glucosidase produced by Humicola grisea var. thermoidea grown on sugarcane bagasse. Journal of Microbiology, 48, 53–62. DOI: 10.1007/s12275-009-0159-x.
Ng, S., Li, C. W., Chan, S. P., Chir, J. L., Chen, P. T., Tong, C. C., Yu, S. M., & Ho, T. H. D. (2010). High-level production of a thermoacidophilic β-glucosidase from Penicillium citrinum YS40-5 by solid-state fermentation with rice bran. Bioresource Technology, 101, 1310–1317. DOI: 10.1016/j.biortech.2009.08.049.
Olofsson, K., Bertilsson, M., & Lidén, G. (2008). A short review on SSF — an interesting process option for ethanol production from lignocellulosic feedstocks. Biotechnology for Biofuels, 1, 1–14. DOI: 10.1186/1754-6834-1-7.
Pei, X. Q., Yi, Z. L., Tang, C. G., & Wu, Z. L. (2011). Three amino acid changes contribute markedly to the thermostability of β-glucosidase BglC from Thermobifida fusca. Bioresource Technology, 102, 3337–3342. DOI: 10.1016/j.biortech.2010.11.025.
Philippoussis, A., Zervakis, G., & Diamantopoulou, P. (2001). Bioconversion of agricultural lignocelulosic wastes through the cultivation of the edible mushrooms Agrocibes aegerita, Volvariella volvacea and Pleurotus spp. World Journal Microbiology & Biotechnology, 17, 191–200. DOI: 10.1023/a:1016685530312.
Qi, B., Wang, L. M., & Liu, X. J. (2009). Purification and characterization of β glucosidase from newly isolated Aspergillus sp. MT-0204. African Journal of Biotechnology, 8, 2367–2374.
Ragauskas, A. J., Williams, C. K., Davison, B. H., Britovsek, G., Cairney, J., Eckert, C. A., Frederick, W. J., Jr., Hallett, J. P., Leak, D. J., Liotta, C. L., Mielenz, J. R., Murphy, R., Templer, R., & Tschaplinski, T. (2006). The path forward for biofuels and biomaterials. Science, 311, 484–489. DOI: 10.1126/science.1114736.
Rajoka, M. I., Akhtar, M. W., Hanif, A., & Khalid, A. M. (2006). Production and characterization of a highly active cellobiase from Aspergillus niger grown in solid state fermentation. World Journal of Microbiology & Biotechnology, 22, 991–998. DOI: 10.1007/s11274-006-9146-0.
Ribeiro, L. F. C., Ribeiro, L. F., Jorge, J. A., & Polizeli, M. L. T. M. (2014). Screening of filamentous fungi for xylanases and cellulases not inhibited by xylose and glucose. British Biotechnology Journal, 4, 30–39. DOI: 10.9734/bbj/2014/6066.
Singhania, R. R., Sukumaran, R. K., Patel, A. K., Larroche, C., & Pandey, A. (2010). Advancement and comparative profiles in the production technologies using solid-state and submerged fermentation for microbial cellulases. Enzyme and Microbiology Technology, 46, 541–549. DOI: 10.1016/j.enzmictec.2010.03.010.
Soni, R., Nazir, A., & Chadha, B. S. (2010). Optimization of cellulase production by a versatile Aspergillus fumigatus fresenius strain (AMA) capable of efficient deinking and enzymatic hydrolysis of Solka floc and bagasse. Industrial Crops and Products, 31, 277–283. DOI: 10.1016/j.indcrop.2009.11.007.
Sonia, K. G., Chadha, E. B. S., Badhan, E. A. K., Saini, H. S., & Bhat, E. M. (2008). Identification of glucose tolerant acid active β-glucosidases from thermophilic and thermotolerant fungi. World Journal Microbiology & Biotechnology, 24, 599–604. DOI: 10.1007/s11274-007-9512-6.
Sørensen, A., Ahring, B. K., Lübeck, M., Ubhayasekera, W., Bruno, K. S., Culley, D. E., & Lübeck, P. S. (2012). Identifying and characterizing the most significant β-glucosidase of the novel species Aspergillus saccharolyticus. Canadian Journal of Microbiology, 58, 1035–1046. DOI: 10.1139/w2012-076.
Souza, F. H. M., Nascimento, C. V., Rosa, J. C., Masui, D. C., Leone, F. A., Jorge, J. A., & Furriel, R. P. M. (2010). Purification and biochemical characterization of a mycelial glucose and xylose stimulated β-glucosidase from the thermophilic fungus Humicola insolens. Process Biochemistry, 45, 272–278. DOI: 10.1016/j.procbio.2009.09.018.
Tamura, K., Stecher, G., Peterson, D., Filipski, A., & Kumar, S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0. Molecular Biology and Evolution, 30, 2725–2729. DOI: 10.1093/molbev/mst197.
Tengborg, C., Galbe, M., & Zacchi, G. (2001). Influence of enzyme loading and physical parameters on the enzymatic hydrolysis of steam pretreated softwood. Biotechnology Progress, 17, 110–117. DOI: 10.1021/bp000145+.
Tu, M. B., Zhang, X., Kurabi, A., Gilkes, N., Mabee, W., & Saddler, J. (2006). Immobilization of β-glucosidase on Eupergit C for lignocelluloses hydrolysis. Biotechnology Letters, 28, 151–156. DOI: 10.1007/s10529-005-5328-3.
Turenne, C. Y., Sanche, S. E., Hoban, D. J., Karlowsky, J. A., & Kabani, A. M. (1999). Rapid identification of fungi by using the ITS2 genetic region and an automated fluorescent capillary electrophoresis system. Journal of Clinical Microbiology, 7, 1846–1851.
Vintila, T., Dragomirescu, M., Croitoriu, V., Vintila, C., Barbu, H., & Sand, C. (2010). Saccharification of lignocelluloses — with reference to Miscanthus — using different cellulases. Romanian Biotechnological Letters, 15, 5498–5504.
White, T. J., Bruns, T., Lee, S., & Taylor, J. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In M. A. Innis, D. H. Gelfand, J. J. Sninsky, & T. J. White (Eds.), PCR Protocols: a guide to methods and applications (pp. 315–322). New York, NY, USA: Academic Press.
Wu, Z., & Lee, Y. Y. (1997). Inhibition of the enzymatic hydrolysis of cellulose by ethanol. Biotechnology Letters, 19, 977–979. DOI: 10.1023/a:1018487015129.
Yang, S. Q., Jiang, Z. Q., Yan, Q. J., & Zhu, H. F. (2008). Characterization of a thermostable extracellular β-glucosidase with activities of exoglucanase and transglycosylation from Paecilomyces thermophila. Journal of Agricultural and Food Chemistry, 56, 602–608. DOI: 10.1021/jf072279.
Zanoelo, F. F., Polizeli, M. L. T. M., Terenzi, H. F., & Jorge, J. A. (2004). β-Glucosidase activity from the thermophilic fungus Scytalidium thermophilum is stimulated by glucose and xylose. FEMS Microbiology Letters, 240, 137–143. DOI: 10.1016/j.femsle.2004.09.021.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Oriente, A., Tramontina, R., de Andrades, D. et al. Characterization of a novel Aspergillus niger beta-glucosidase tolerant to saccharification of lignocellulosic biomass products and fermentation inhibitors. Chem. Pap. 69, 1050–1057 (2015). https://doi.org/10.1515/chempap-2015-0111
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1515/chempap-2015-0111