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How to Cite Complete Issue More Information About This Article Journal's Webpage in Redalyc.Org Scientific Information System Re Revista de Biología Tropical ISSN: 0034-7744 ISSN: 2215-2075 Universidad de Costa Rica Martín Giorgio, Ernesto; Villalba, Laura Lidia; Lucio Robledo, Gerardo; Zapata, Pedro Darío; Nazareno Saparrat, Mario Carlos Cellulolytic ability of a promising Irpex lacteus (Basidiomycota: Polyporales) strain from the subtropical rainforest of Misiones province, Argentina Revista de Biología Tropical, vol. 66, no. 3, 2018, July-September, pp. 1034-1045 Universidad de Costa Rica DOI: https://doi.org/10.15517/rbt.v66i3.30589 Available in: https://www.redalyc.org/articulo.oa?id=44959350007 How to cite Complete issue Scientific Information System Redalyc More information about this article Network of Scientific Journals from Latin America and the Caribbean, Spain and Journal's webpage in redalyc.org Portugal Project academic non-profit, developed under the open access initiative Cellulolytic ability of a promising Irpex lacteus (Basidiomycota: Polyporales) strain from the subtropical rainforest of Misiones province, Argentina Ernesto Martín Giorgio1*, Laura Lidia Villalba1, Gerardo Lucio Robledo2,3, Pedro Darío Zapata1 & Mario Carlos Nazareno Saparrat4,5 1. Laboratorio de Biotecnología Molecular. Instituto de Biotecnología Misiones “Dra. María Ebe Reca”. Universidad Nacional de Misiones (UNaM). Ruta 12 Km 7 1/2. Campus Universitario, CP 3300. Posadas, Misiones, Argentina; [email protected], [email protected], [email protected] 2. Laboratorio de Micología. Instituto Multidisciplinario de Biología Vegetal-CONICET. Universidad Nacional de Córdoba (UNC). CC 495, CP 5000, Córdoba, Argentina; [email protected] 3. Fundación FungiCosmos. Av. General Paz 154, 4º piso, oficina 4, Córdoba, Argentina. 4. Instituto de Fisiología Vegetal (INFIVE) CCT-La Plata-Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)-Universidad Nacional de La Plata (UNLP). CC 327, CP 1900, La Plata, Argentina; [email protected] 5. Instituto de Botánica Spegazzini. Facultad de Ciencias Naturales y Museo. UNLP. La Plata, Argentina. * Correspondence Received 29-I-2018. Corrected 11-V-2018. Accepted 13-VI-2018. Abstract: The cellulolytic activity of fungi growing in the subtropical rainforest of Misiones (Argentina) rep- resents a challenge in the technological development of the production of cellulosic bioethanol in the region using native sources. These fungi are promising to obtain sustainable enzyme cocktails using their enzymes. Cellulolytic ability of 22 white-rot fungi isolated from the subtropical rainforest of Misiones-Argentina in agar medium with two types of cellulosic substrates, carboxy-methylcellulose or crystalline cellulose, were com- paratively analyzed, and the activity of two cellulolytic enzymes was evaluated in liquid medium. Although all isolates were able to grow and degrade both substrates in agar medium, and to produce total cellulase Filter paper (FPase) and endo-β-1,4-glucanase (EG) activities in broth, the isolate Irpex sp. LBM 034 showed the greatest enzymatic levels (FPase, 65.45 U L-1; EG, 221.21 U L-1). Therefore, the ITS sequence of this fungus was sequenced and analyzed through a phylogenetic analysis. These results indicate that the isolate LBM 034, corre- sponding to Irpex lacteus, has a promising cellulolytic ability and enzymes such as EG useful in sustainable sac- charification of cellulosic materials in the region. Rev. Biol. Trop. 66(3): 1034-1045. Epub 2018 September 01. Key words: white-rot fungi; Irpex lacteus; cellulases; FPase; endo-β-1; 4-glucanase. Cellulose is considered to be the most and 15 000 for native cellulose cotton (Habi- abundant organic compound derived from plant bi, Lucia, & Rojas, 2010; Siqueira, Bras, & biomass. Perpetual renewal of plant biomass Dufresne, 2010). via the process of photosynthesis ensures an Cellulolytic enzymes degrade cellulose inexhaustible supply of such material. The by cleaving the glycosidic bonds. Many fungi worldwide production of this biopolymer is and bacteria are capable of producing multiple estimated to be between 1010 and 1011 tons enzymes, collectively known as cellulases, each year (Lynd, Weimer, van Zyl, & Pretorius, that act in a synergistic manner to hydro- 2002). Cellulose is a linear homopolysaccha- lyze the β-1,4-D-glycosidic bonds within the ride of β-1.4-linked anhydro-D-glucose units cellulose molecule (Akiba, Kimura, Yama- with a degree of polymerization of approxi- moto, & Kumagai, 1995). Cellulases can be mately 10 000 for cellulose chains in nature classified into three types: endoglucanases 1034 Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 66(3): 1034-1045, September 2018 (endo-β-1,4-glucanase, EC 3.2.1.4), cellobiohy- Lechner, & Popoff, 2008), there is no data for drolases (cellulose β-1.4-cellobiosidase, EC the rest of the region that remains still under- 3.2.1.91), and β-glucosidases (β-D-glucoside explored (Niveiro, Popoff, & Albertó, 2010). glucohydrolase, EC 3.2.1.21). Endogluca- A high forestry activity is found in the nases randomly hydrolyze internal β-1,4-D- province of Misiones-Argentina due to avail- glycosidic bonds in cellulose. As a result, the ability of a lot of plant sources. Therefore, there polymer rapidly decreases in length and the is an increase of woody wastes (Teza et al., concentration of the reducing sugar increases 2012). In spite of the huge amount of this plant slowly. Cellobiohydrolases hydrolyze cellulose material, there are still burning practices for its by removing the cellobiose units from the non- use, however the law XVI N°106 since 2012 reducing end of cellulose, and β-glucosidases, prohibits this activity and promotes their utili- cleave glucose units from cellooligosaccha- zation in sustainable processes (Sistema Argen- rides (Siddiqui, Shemsi, Anwar, Rashid, & tino de Información Jurídica, 2012). This has Rajoka, 1999). lead to the search for innovative biotechnologi- In Argentina, subtropical and tropical rain- cal strategies such as the production of second forests are restricted to small areas that do generation biofuels, which is also depending not reach 2 % of the land area, and yet, are upon reactive and enzymes for the process. In home to more than 50 % of the biodiver- this sense, the availability of enzyme cocktails sity (Brown, Grau, Lomáscolo, & Gasparri, from autochthonous fungal sources is also a 2002). Specifically these forests belong to priority (King, Donnelly, Bergstrom, Walker, two phytogeographic regions of the Amazon & Gibson, 2008). Domain: Yungas rainforests, located on the In the present study, we analyzed and com- eastern slopes of the foothills in the Northwest pared the cellulolytic ability of 22 white-rot of the country; and the Paranaense rainforest fungi, isolated from woody materials collected located in the province of Misiones and North- from the subtropical rainforest of the province east of Corrientes (Cabrera, 1994). Currently, of Misiones, Argentina, grown on agar medium studies of wood-decaying fungi diversity are supplemented with either sodium carboxy- in an active process of analysis and dissemina- methylcellulose (CMC) or crystalline cellu- tion (Rajchenberg & Robledo, 2013; Gallo, lose (CC). Furthermore, submerged cultures Robledo, Romero, & Catania, 2014; Robledo, of these fungi in a liquid medium with CMC Giorgio, Franco, Popoff, & Decock, 2014). were conducted to select the isolate with the However, knowledge about the cellulo- highest level of filter paper assay (FPase) and lytic enzyme production of these fungi is very endo-β-1,4-glucanase (EG) activities, which much restricted. The forest environments in the was subsequently identified at molecular level. province of Misiones in Northeast Argentina, are a rich reservoir of biodiversity (Zuloaga, MATERIALS AND METHODS Morrone, & Rodríguez, 1999). Exploration and description of new fungi from Paranaense rain- Fungal isolates: Fungal isolates from dif- forest can reveal innovative abilities for poten- ferent sources were selected for this study tial biotechnological applications such as plant (Table 1). They belong to the following culture biomass conversion (Saparrat, Martínez, Cabel- collections: 1. Facultad de Ciencias Exactas lo, & Arambarri, 2002; Giorgio et al., 2012; y Naturales, Universidad Nacional de Buenos Daâssi et al., 2016). Moreover, the relevance of Aires (FCEyN-UBA); 2. Instituto Multidiscipli- screening of new isolates in addition to culture nario de Biología Vegetal (IMBIV-CONICET) collection strains has been already highlighted and 3. Instituto de Biotecnología Misiones, (Tekere, Mswaka, Zvauya, & Read, 2001). In Universidad Nacional de Misiones (InBioMis- this sense, although most reports available are UNaM). Stock cultures of the Basidiomycetes related to the Iguazú National Park (Wright, isolates tested were kept at 4 °C on 20 (g L-1) Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 66(3): 1034-1045, September 2018 1035 TABLE 1 Fungal isolates used in this work and their source of conservation Culture Collection Fungal isolates Colección de Cultivos de Hongos, Coriolus versicolor (L.) Quél. BAFC 266 Departamento de Biodiversidad y Biología Experimental, Irpex lacteus (Fr.) Fr. BAFC 1171 FCEyN, UBA Lenzites elegans (Spreng.) Pat. BAFC 2127 Phlebia brevispora Nakasone BAFC 633 Pycnoporus sanguineus (L.) Murrill BAFC 2126 Trametes villosa (Sw.) Kreisel BAFC 2755 Laboratorio de Biotecnología Molecular, InBioMis Trametes sp. LBM 029 Trametes sp. LBM 030 Trametes sp. LBM 031 Trametes sp. LBM 033 Trametes villosa (Sw.) Kreisel LBM 109 Trametes
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