Potential of the Aspergillus labruscus ITAL 22.223 as a producer of cellulolytic enzymes and xylanase under solid-state fermentation
Keywords:Aspergillus, Cellulase, Solid-state fermentation, Xylanase
Background: The enzymatic hydrolysis of the lignocellulosic biomass to obtain saccharides that can be used for the production of bioethanol is an important field in the renewable energy area. For this purpose, fungal cellulases and xylanases can be applied.
Methods: Aspergillus labruscus ITAL 22.223 was cultured under SSF with agroindustrial residues and by-products as substrates, humidified with different moistening agents, at different proportions (1:0.5, 1:1, 1:1.5 and 1:2; m/v), for different periods (24-216 h) at 25ºC. The extract obtained was used for determination of the cellulase and xylanase activities. The influence of temperature, pH and different compounds on xylanase activity was analyzed.
Results: A. labruscus produced cellulases and xylanase under solid-state fermentation (SSF) using agroindustrial by products and residues as carbon source/substrates. The best production of β-glucosidase (6.3 U/g of substrate) was obtained in the presence of rye bran, whereas for the CMCase it was in the presence of crushed soybean (5.1 U/g of substrate) and xylanase using oat bran (74.8 U/g of substrate) as substrates, for 168 h of cultivation at 25ºC. Considering the high xylanase production, the best moistening agent and its proportion (tap water, 1:2 m/v) were determined. Optimum of temperature and pH for xylanase activity was determined as 55ºC and pH 5.5. The xylanase activity was inhibited by different salts, with exception of MnSO4. It was also inhibited by organic solvents, detergents, EDTA, urea and β-mercaptoethanol.
Conclusions: The fungus A. labruscus presented potential to produce enzymes from the cellulolytic complex and xylanase using low cost substrates.
Pandey A, Soccol CR, Mitchell D. New developments in solid-state fermentation: I-bioprocesses and products. Process Biochem. 2000;35:1153-69.
Silva NLC, Betancur GJV, Vasquez MP, Gomes EB, Pereira Jr N. Ethanol production from residual wood chips of cellulose industry: acid pretreatment investigation, hemicellulosic hydrolysate fermentation, and remaining solid fraction fermentation by SSF process. Appl Biochem Biotecnhol 2011;163:928-36.
Dillon AJP. Celulases. In: Enzimas como agentes biotecnológicos; Said S, Pietro RCLR (Org). Ribeirão Preto: Legis Summa, 2004: 243-269.
Ferreira-Filho EX. Xilanases In: Enzimas como agentes biotecnológicos; Said S, Pietro RCLR (Org). Ribeirão Preto: Legis Summa; 2004: 137-148.
Carvalho W, Canilha L, Ferraz A, Milagres AMF. Uma visão sobre a estrutura, composição e biodegradação da madeira. Quim Nova. 2009;32:2191-5.
Aguiar CM, Lucena SL. Produção de celulases por Aspergillus niger e cinética da desativação celulásica. Acta Sci Technol. 2011;33:385-91.
Fungaro M, Ferranti L, Massi F, Silva J, Sartori D, Taniwaki M, Frisvad J, Iamanaka B. Aspergillus labruscus sp. nov., a new species of Aspergillus section Nigri discovered in Brazil. Sci Rep. 2017;7:1-9.
Vogel HJ. Distribution of lysine pathways among fungi: evolutionary implications. Am Nat. 1964;98:435-446.
Khanna P, Sundari SS, Kumar NJ. Production, isolation and partial purification of xylanases from an Aspergillus sp. World J Microbiol Biotechnol. 1995;11:242-3.
Rizzatti ACS, Jorge JA, Terenzi HF, Rechia CG, Polizeli MLTM. Purification and properties of a thermostable extracellular beta-D-xylosidase produced by a thermotolerant Aspergillus phoenicis. J Ind Microbiol Biotechnol. 2001;3:156-60.
Ghose TK. Measurement of cellulase activities. Pure Appl Chem. 1987;59:257-268.
Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem. 1959;31:426.
Coston MB, Loomis WF. Isozymes of β-glucosidase in Dictyostelium discoideum. J Bacteriol. 1969;100:1208-17.
Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248-54.
Quiroz RDLC, Roussos S, Hernández D, Rodríguez R, Castillo F, Aguilar CN. Challenges and opportunities of the bio-pesticides production by solid-state fermentation: filamentous fungi as model. Crit Rev Biotechnol. 2015;35:326-33.
Yazid NA, Barrena R, Komilis D, Sánchez A. Solid-State fermentation as a novel paradigm for organic waste valorization: a review. Sustainability 2017;9:224-52.
Liu Z, Fatehi P, Sadeghi S, Ni Y. Application of hemicelluloses precipitated via ethanol treatment of pre-hydrolysis liquor in high-yield poulp. Bioresour Technol. 2011;102:9613-8.
Zúñiga UFR, Farinas CS, Neto VB, Couri S. Produção de celulases por Aspergillus niger por Fermentação em Estado Sólido. Pesq Agropec Bras. 2011;46:912-9.
Yamane Y, Fujita J, Shimizu R, Hiyoshi A, Fukuda H, Kizaki Y, Wakabayashi S. Production of cellulose and xylan-degrading enzymes by a koji mold, Aspergillus oryzae, and their contribution to the maceration of rice endosperm cell wall. J Biosci Bioeng. 2002;93:9-14.
Sherief AA, El-Tanash AB, Atia N. Cellulase production by Aspergillus fumigatus grown on mixed substrate of rice straw and wheat bran. Res J Microbiol. 2010;5:199-211.
Betini JHA, Michelin M, Peixoto-Nogueira SC, Jorge JA, Terenzi HF, Polizeli MLTM. Xylanases from Aspergillus niger, Aspergillus niveus and Aspergillus ochraceus produced under solid-state fermentation and their application in cellulose pulp bleaching. Biopr Biosys Eng. 2009;32:819-24.
Carmona EC, Pizzirani-Kleiner AA, Monteiro RTR, Jorge JA. Xylanase production by Aspergillus versicolor. J Basic Microbiol. 1997;37:387-94.
Farinas CS, Loyo MM, Baraldo Jr A, Tardioli PW, Bertucci-Neto V, Couri S. Finding stable cellulase and xylanase: evaluation of the synergistic effect on pH and temperature. New Biotechnol. 2010;27:810-5.
Ahmad Z, Butt MS, Riaz M. Partial purification and characterization of xylanase produced from Aspergillus niger using wheat bran. Pak J Agr Sci. 2012;50:433-7.
Yegin S. Single-step purification and characterization of an extreme halophilic, ethanol tolerant and acidophilic xylanase from Aureobasidium pullulans NRRL Y-2311-1 with application potential in food industry. Food Chem. 2017;221:67-75.
Maitain-Alfenas GP, Oliveira MB, Nagem RAP, Vries RP, Guimarães VM. Characterization and biotechnological application of recombinant xylanases from Aspergillus nidulans. Int J Biol Macromol. 2016;91:60-7.
Do TT, Quyen DT, Nguyen TN, Nguyen VT. Molecular characterization of a glycosil hydrolase family 10 xylanase from Aspergillus niger. Prot Expres Purif. 2013;92:196-202.
Sandrim VC, Rizzatti ACS, Terenzi HF, Jorge JA, Milagres AMF, Polizeli MLTM. Purification and biochemical characterization of two xylanases produced by Aspergillus caespitosus and the potential for kraft pulp bleaching. Process Biochem. 2005;40:1823-8.
Hmida-Sayari A, TakTek S, Elgharbi F, Bejar S. Biochemical characterization, cloning and molecular modeling of a detergent and organic solvent stable family 11 xylanase from the newly isolated Aspergillus niger US368 strain. Process Biochem. 2012;47:1839-47.