Assessment of bacterial and fungal (hemi)cellulose-degrading enzymes in saccharification of ammonia fibre expansion-pretreated Arundo donax.

Simona Giacobbe,Venkatesh Balan,Mauro Mori,Vincenza Faraco,Salvatore Montella,Massimo Fagnano

doi:10.1007/s00253-015-7066-3

Abstract

This study reports enzymatic hydrolysis of the biomass of the giant reed (Arundo donax L.) after ammonia fibre expansion (AFEX) pretreatment. In particular, the capacity of the arabinofuranosidase from the fungus Pleurotus ostreatus recombinantly expressed in Pichia pastoris rPoAbf, its evolved mutant rPoAbf F435Y/Y446F and the endo-cellulase from Streptomyces sp. G12 CelStrep recombinantly expressed in Escherichia coli to enhance the hydrolysis of AFEX-treated A. donax was investigated, using the corn stover as reference feedstock. The investigated enzymes were assayed using a mixture of purified cellulases (CBHI, CBHII, EGI and βG), endoxylanases (LX3, LX4) and accessory hemicellulases (LarbF and LβX) as reference enzyme mixture and substituting EGI with rCelStrep and LarbF with rPoAbf or rPoAbf F435Y/Y446F. The use of rPoAbf F435Y/Y446F in the substitution of LarbF led to improvements in sugar conversion, giving a glucan, xylan and arabinan conversion after 72 h of around 62, 63 and 80 %, respectively, similar or higher than those (44, 66 and 55 %) achieved by 72 h hydrolysis with commercial enzymes Novozymes Cellic®, Ctec3 and Htec3. The enzymes rPoAbf, rPoAbf F435Y/Y446F and rCelStrep were also investigated for their effect on hydrolysis of AFEX-pretreated A. donax by addition to commercial enzyme mixture Novozymes Cellic®, Ctec3 and Htec3, and it was shown that the addition of rPoAbf and its evolved mutant rPoAbf F435Y/Y446F enhanced both xylan and arabinan conversions, which achieved 80 % after 6 days of saccharification with rPoAbf F435Y/Y446F.

Highlights

The policies for climate change mitigation promote the replacement of fossil fuels and petroleum-based products with alternative bioproducts from renewable resources such as biomass crops (Kajaste 2014)
This study was aimed at evaluating the conversion of A. donax into monosaccharides for bioethanol or bioproducts production by enzymatic hydrolysis of the polysaccharides issued from pretreatment of the biomass by ammonia fibre expansion (AFEX)
The AFEX-tested conditions, condition 1, condition 2 and condition 3 reported in Table 3, were chosen since they had been previously identified as the best conditions for the AFEX pretreatment of Miscanthus x giganteus (Murnen et al 2007), corn stover (Balan et al 2009b) and switchgrass (Bals et al 2010), respectively

Summary

Introduction

The policies for climate change mitigation promote the replacement of fossil fuels and petroleum-based products with alternative bioproducts from renewable resources such as biomass crops (Kajaste 2014). In Mediterranean environments, hilly areas are considered not appropriate for the traditional cereal production because yield and gross income are very low and because the traditional cropping system (deep soil tillage at the end of August and sowing in November) causes extreme vulnerability to soil erosion (Diodato et al 2009, 2011; Fagnano et al 2012). In these conditions, perennial biomass crops such as the giant reed (Arundo donax L.) proved to reduce soil erosion and to increase the potential gross income of farmers (Fagnano et al. Appl Microbiol Biotechnol (2016) 100:2213–2224

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