Abstract
From three cell-associated β-xylosidases produced by Aureobasidium pullulans CBS 135684, the principal enzyme was enriched to apparent homogeneity and found to be active at high temperatures (60–70 °C) over a pH range of 5–9 with a specific activity of 163.3 units (U) mg−1. The enzyme was thermostable, retaining over 80% of its initial activity after a 12-h incubation at 60 °C, with half-lives of 38, 22, and 10 h at 60, 65, and 70 °C, respectively. Moreover, it was tolerant to xylose inhibition with a K i value of 18 mM. The K m and V max values against p-nitrophenyl-β-d-xylopyranoside were 5.57 ± 0.27 mM and 137.0 ± 4.8 μmol min−1 mg−1 protein, respectively. When combining this β-xylosidase with xylanase from the same A. pullulans strain, the rate of black liquor xylan hydrolysis was significantly improved by up to 1.6-fold. The maximum xylose yield (0.812 ± 0.015 g g−1 dry weight) was obtained from a reaction mixture containing 10% (w/v) black liquor xylan, 6 U g−1 β-xylosidase and 16 U g−1 xylanase after incubation for 4 h at 70 °C and pH 6.0.
Similar content being viewed by others
References
Hu, J., Davies, J., Mok, Y. K., Gene, B., Lee, Q. F., Arato, C., & Saddler, J. N. (2016). Enzymatic hydrolysis of industrial derived xylo-oligomers to monomeric sugars for potential chemical/biofuel production. ACS Sustainable Chemistry & Engineering, 4, 7130–7136.
Quiñones, T. S., Retter, A., Hobbs, P. J., Budde, J., Heiermann, M., Plöchl, M., & Ravella, S. R. (2015). Production of xylooligosaccharides from renewable agricultural lignocellulose biomass. Biofuels, 6, 147–155.
Lisboa, S. A., Evtuguin, D. V., Neto, C. P., & Goodfellow, B. J. (2005). Isolation and structural characterization of polysaccharides dissolved in Eucalyptus globulus kraft black liquors. Carbohydrate Polymers, 60, 77–85.
Bankeeree, W., Prasongsuk, S., Imai, T., Lotrakul, P., & Punnapayak, H. (2016). A novel xylan-polyvinyl alcohol hydrogel bead with laccase entrapment for decolorization of reactive black 5. BioResources, 11, 6984–7000.
Kumar, K. S., Arumugam, N., Permaul, K., & Singh, S. (2016). Thermostable enzymes and their industrial applications. Microbial Biotechnology: An Interdisciplinary Approach, Taylor and Francis, CRC Press, Boca Raton, Florida. Chapter 5, pp. 115–162.
Shao, W., Xue, Y., Wu, A., Kataeva, I., Pei, J., Wu, H., & Wiegel, J. (2011). Characterization of a novel β-Xylosidase, XylC, from Thermoanaerobacterium saccharolyticum JW/SL-YS485. Applied and Environmental Microbiology, 77, 719–726.
Leathers, T. D. (1986). Color variants of Aureobasidium pullulans overproduce xylanase with extremely high specific activity. Applied and Environmental Microbiology, 52, 1026–1030.
Manitchotpisit, P., Leathers, T. D., Peterson, S. W., Kurtzman, C. P., Li, X. L., Eveleigh, D. E., Lotrakul, P., Prasongsuk, S., Dunlap, C. A., Vermillion, K. E., & Punnapayak, H. (2009). Multilocus phylogenetic analyses, pullulan production and xylanase activity of tropical isolates of Aureobasidium pullulans. Mycological Research, 113, 1107–1120.
Bankeeree, W., Lotrakul, P., Prasongsuk, S., Chaiareekij, S., Eveleigh, D. E., Kim, S. W., & Punnapayak, H. (2014). Effect of polyols on thermostability of xylanase from a tropical isolate of Aureobasidium pullulans and its application in prebleaching of rice straw pulp. SpringerPlus, 3, 37–48.
Terrasan, C. R. F., Guisan, J. M., & Carmona, E. C. (2016). Xylanase and β-xylosidase from Penicillium janczewskii: purification, characterization and hydrolysis of substrates. Electronic Journal of Biotechnology, 23, 54–62.
Li, H., Liu, J., Wu, J., Xue, Y., Gan, L., & Long, M. (2014). Comparative analysis of enzymatic hydrolysis of miscanthus xylan using Aspergillus niger, Hypocrea orientalis, and Trichoderma reesei xylan-degrading enzymes. BioResources, 9, 2191–2202.
Bankeeree, W., Lotrakul, P., Prasongsuk, S., Kim, S. W., & Punnapayak, H. (2016). Enhanced production of cellulase-free thermoactive xylanase using corncob by a black yeast, Aureobasidium pullulans CBS 135684. Korean Chemical Engineering Research, 54, 822–829.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., & Randall, R. J. (1951). Protein measurement with the folin phenol reagent. Journal of Biological Chemistry, 193, 265–275.
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685.
Miller, G. L. (1959). Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry, 31, 426–428.
Lemmnitzer, K., Süß, R., & Schiller, J. (2015). TLC/MALDI MS of carbohydrates planar chromatography—mass spectrometry (pp. 327–344). Boca Raton, Florida: CRC Press.
Box, G. E., & Behnken, D. W. (1960). Some new three level designs for the study of quantitative variables. Technometrics, 2, 455–475.
Iembo, T., Da Silva, R., Pagnocca, F. C., & Gomes, E. (2002). Production, characterization, and properties of β-glucosidase and β-xylosidase from a strain of Aureobasidium sp. Applied Biochemistry and Microbiology, 38, 549–552.
Ohta, K., Fujimoto, H., Fujii, S., & Wakiyama, M. (2010). Cell-associated β-xylosidase from Aureobasidium pullulans ATCC 20524: Purification, properties, and characterization of the encoding gene. Journal of Bioscience and Bioengineering, 110, 152–157.
Dobberstein, J., & Emeis, C. C. (1991). Purification and characterization of β-xylosidase from Aureobasidium pullulans. Applied Microbiology and Biotechnology, 35, 210–215.
Jain, I., Kumar, V., & Satyanarayana, T. (2014). Applicability of recombinant β-xylosidase from the extremely thermophilic bacterium Geobacillus thermodenitrificans in synthesizing alkylxylosides. Bioresource Technology, 170, 462–469.
Semenova, M. V., Drachevskaya, M. I., Sinitsyna, O. A., Gusakov, A. V., & Sinitsyn, A. P. (2009). Isolation and properties of extracellular β-xylosidases from fungi Aspergillus japonicus and Trichoderma reesei. Biochemistry (Moscow), 74, 1002–1008.
Díaz-Malváez, F. I., García-Almendárez, B. E., Hernández-Arana, A., Amaro-Reyes, A., & Regalado-González, C. (2013). Isolation and properties of β-xylosidase from Aspergillus niger GS1 using corn pericarp upon solid state fermentation. Process Biochemistry, 48, 1018–1024.
Kirikyali, N., Wood, J., & Connerton, I. F. (2014). Characterisation of a recombinant β-xylosidase (xylA) from Aspergillus oryzae expressed in Pichia pastoris. AMB Express, 4, 68–74.
Saha, B. C. (2003). Purification and properties of an extracellular β-xylosidase from a newly isolated Fusarium proliferatum. Bioresource Technology, 90, 33–38.
Iembo, T., Azevedo, M. O., Bloch Jr., C., & Filho, E. X. F. (2006). Purification and partial characterization οf a new β-xylosidase from Humicola grisea var. thermoidea. World Journal of Microbiology and Biotechnology, 22, 475–479.
Xia, W., Shi, P., Xu, X., Qian, L., Cui, Y., Xia, M., & Yao, B. (2015). High level expression of a novel family 3 neutral β-xylosidase from Humicola insolens Y1 with high tolerance to d-xylose. PloS One, 10, e0117578.
Knob, A., & Carmona, E. C. (2012). Purification and properties of an acid β-xylosidase from Penicillium sclerotiorum. Annals of Microbiology, 62, 501–508.
Teng, C., Jia, H., Yan, Q., Zhou, P., & Jiang, Z. (2011). High-level expression of extracellular secretion of a β-xylosidase gene from Paecilomyces thermophila in Escherichia coli. Bioresource Technology, 102, 1822–1830.
Nieto-Domínguez, M., de Eugenio, L. I., Barriuso, J., Prieto, A., de Toro, B. F., Canales-Mayordomo, Á., & Martínez, M. J. (2015). Characterization of a novel pH-stable GH3 β-xylosidase from Talaromyces amestolkiae: an enzyme displaying regioselective transxylosylation. Applied and Environmental Microbiology, 81, 6380–6392.
Lenartovicz, V., de Souza, C. G. M., Moreira, F. G., & Peralta, R. M. (2003). Temperature and carbon source affect the production and secretion of a thermostable β-xylosidase by Aspergillus fumigatus. Process Biochemistry, 38, 1775–1780.
Yan, Q. J., Wang, L., Jiang, Z. Q., Yang, S. Q., Zhu, H. F., & Li, L. T. (2008). A xylose-tolerant β-xylosidase from Paecilomyces thermophila: characterization and its co-action with the endogenous xylanase. Bioresource Technology, 99, 5402–5410.
Singh, S. M., Cabello-Villegas, J., Hutchings, R. L., & Mallela, K. M. G. (2010). Role of partial protein unfolding in alcohol-induced protein aggregation. Proteins: Structure, Function, and Bioinformatics, 78, 2625–2637.
Katapodis, P., Nerinckx, W., Claeyssens, M., & Christakopoulos, P. (2006). Purification and characterization of a thermostable intracellular β-xylosidase from the thermophilic fungus Sporotrichum thermophile. Process Biochemistry, 41, 2402–2409.
Terrasan, C. R., Temer, B., Sarto, C., Silva, J. F. G., & Carmona, E. C. (2013). Xylanase and β-xylosidase from Penicillium janczewskii: production, physico-chemical properties, and application of the crude extract to pulp biobleaching. BioResources, 8, 1292–1305.
Jordan, D. B., & Braker, J. D. (2007). Inhibition of the two-subsite β-d-xylosidase from Selenomonas ruminantium by sugars: competitive, noncompetitive, double binding, and slow binding modes. Archives of Biochemistry and Biophysics, 465, 231–246.
Jönsson, A.-S., Nordin, A.-K., & Wallberg, O. (2008). Concentration and purification of lignin in hardwood kraft pulping liquor by ultrafiltration and nanofiltration. Chemical Engineering Research and Design, 86, 1271–1280.
Kambourova, M., Mandeva, R., Fiume, I., Maurelli, L., Rossi, M., & Morana, A. (2007). Hydrolysis of xylan at high temperature by co-action of the xylanase from Anoxybacillus flavithermus BC and the β-xylosidase/α-arabinosidase from Sulfolobus solfataricus Oα. Journal of Applied Microbiology, 102, 1586–1593.
Acknowledgements
This research was performed under the Core-to-Core Program, which was financially supported by the Japan Society for the Promotion of Science (JSPS), National Research Council of Thailand (NRCT), Vietnam Ministry of Science and Technology (MOST), the National University of Laos, Beuth University of Applied Sciences, and Brawijaya University. In addition, financial support from the Rachadapisek Somphot Endowment under Outstanding Research Performance Program and the Rachadapisek Sompote Fund for Postdoctoral Fellowship, Chulalongkorn University, are acknowledged.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of Interest
The authors declare that they have no conflict of interest.
Electronic supplementary material
ESM 1
(DOCX 53 kb)
Rights and permissions
About this article
Cite this article
Bankeeree, W., Akada, R., Lotrakul, P. et al. Enzymatic Hydrolysis of Black Liquor Xylan by a Novel Xylose-Tolerant, Thermostable β-Xylosidase from a Tropical Strain of Aureobasidium pullulans CBS 135684. Appl Biochem Biotechnol 184, 919–934 (2018). https://doi.org/10.1007/s12010-017-2598-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12010-017-2598-x