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Cellulolytic Activities of a Novel Fomitopsis sp. and Aspergillus tubingensis isolated from Philippine Mangroves

Christine Jurene O. Bacal1,2 and Eizadora T. Yu1,*

1Institute of Chemistry, 2Natural Sciences Research Institute,
University of the Philippines, Diliman, Quezon City, Philippines 1101

*Corresponding author: This email address is being protected from spambots. You need JavaScript enabled to view it.

 

ABSTRACT
The ability to deconstruct plant cell wall polysaccharides is inherent in fungal endophytes. As such, discovering organisms that secrete potent cocktails of carbohydrate-active enzymes may hold the key to deconstructing waste agricultural biomass for industrial applications. Based on CMC-Congo red plate based assay, two fungal isolates derived from mangrove trees (JB10 and JB11) showed high enzymatic indices (as high as 5.6 ± 0.18 for JB10). Both isolates were then grown in potato dextrose (PD), carboxymethylcellulose(CMC), and beechwood xylan (XY), and the corresponding endoglucanase, xylanase, and β-glucosidase activities of the enzymes present in crude culture supernatants were determined. JB11 showed significant increase in endoglucanase activity (0.36 ± 0.04 U/mL) in PD, while JB10 endoglucanase activity was similar between the three media. Interestingly, xylanase activity of both isolates was relatively high (ranging 0.26-1.0 U/mL), with JB10 xylanase activity five-fold higher in PD. Lastly, there was 2-4 fold increase detected in β-glucosidase activities (0.59-0.8 U/mL) in both isolates when grown in CMC or XY media. Phylogenetic analysis of the ITS sequences show that JB11 is Aspergillus tubingensis, while JB10 is a novel Fomitopsis sp. isolate.

 

INTRODUCTION
Lignocellulosic material, in the form of agricultural waste, is a promising biofuel feedstock or source of precursors for fine chemicals. A major obstacle in their utilization is their inherent complexity and recalcitrance to degradation. Current pretreatment methods utilizing harsh chemicals have adverse environmental effects and may preclude use of degradation products in downstream chemical synthesis (Eggeman & Elander 2005). Methods based on mild enzymatic hydrolysis of biomass have been employed and is increasingly being viewed as a greener alternative or complement to existing industrial processes (Mosier et al. 2005). Developing optimized enzyme cocktail formulations that increase biomass conversion rates and maximize yields are also being intensely studied (Mohanram et al. 2013).
Fungi have the ability to express and secrete a cocktail of extracellular hydrolytic and oxidative enzymes that act synergistically to deconstruct lignocellulose into its components (Hatakka and Hammel 2010). The fungal proteins secreted into the media are primarily carbohydrate-active enzymes or CAZys. The CAZys are a diverse family of enzymes responsible for assembly (e.g., glycosyltransferases) and breakdown (e.g., glycosyl hydrolases and carbohydrate esterases) of complex carbohydrates (Lombard et al. 2014). For example, the deconstruction of cellulose requires the concerted effort of enzymes belonging to 12 different glycosyl hydrolase families (Lombard et al. 2014). . . . . read more


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