Abstract

BackgroundEthanolic fermentation of lignocellulosic biomass is a sustainable option for the production of bioethanol. This process would greatly benefit from recombinant Saccharomyces cerevisiae strains also able to ferment, besides the hexose sugar fraction, the pentose sugars, arabinose and xylose. Different pathways can be introduced in S. cerevisiae to provide arabinose and xylose utilisation. In this study, the bacterial arabinose isomerase pathway was combined with two different xylose utilisation pathways: the xylose reductase/xylitol dehydrogenase and xylose isomerase pathways, respectively, in genetically identical strains. The strains were compared with respect to aerobic growth in arabinose and xylose batch culture and in anaerobic batch fermentation of a mixture of glucose, arabinose and xylose.ResultsThe specific aerobic arabinose growth rate was identical, 0.03 h-1, for the xylose reductase/xylitol dehydrogenase and xylose isomerase strain. The xylose reductase/xylitol dehydrogenase strain displayed higher aerobic growth rate on xylose, 0.14 h-1, and higher specific xylose consumption rate in anaerobic batch fermentation, 0.09 g (g cells)-1 h-1 than the xylose isomerase strain, which only reached 0.03 h-1 and 0.02 g (g cells)-1h-1, respectively. Whereas the xylose reductase/xylitol dehydrogenase strain produced higher ethanol yield on total sugars, 0.23 g g-1 compared with 0.18 g g-1 for the xylose isomerase strain, the xylose isomerase strain achieved higher ethanol yield on consumed sugars, 0.41 g g-1 compared with 0.32 g g-1 for the xylose reductase/xylitol dehydrogenase strain. Anaerobic fermentation of a mixture of glucose, arabinose and xylose resulted in higher final ethanol concentration, 14.7 g l-1 for the xylose reductase/xylitol dehydrogenase strain compared with 11.8 g l-1 for the xylose isomerase strain, and in higher specific ethanol productivity, 0.024 g (g cells)-1 h-1 compared with 0.01 g (g cells)-1 h-1 for the xylose reductase/xylitol dehydrogenase strain and the xylose isomerase strain, respectively.ConclusionThe combination of the xylose reductase/xylitol dehydrogenase pathway and the bacterial arabinose isomerase pathway resulted in both higher pentose sugar uptake and higher overall ethanol production than the combination of the xylose isomerase pathway and the bacterial arabinose isomerase pathway. Moreover, the flux through the bacterial arabinose pathway did not increase when combined with the xylose isomerase pathway. This suggests that the low activity of the bacterial arabinose pathway cannot be ascribed to arabitol formation via the xylose reductase enzyme.

Highlights

  • Ethanolic fermentation of lignocellulosic biomass is a sustainable option for the production of bioethanol

  • Construction of arabinose and xylose fermenting strains TMB3075 (XR/xylitol dehydrogenase (XDH) strain) and TMB3076 (XI strain) Two strains, harbouring a chromosomally integrated, bacterial arabinose utilisation pathway [26] that consists of Larabinose isomerase (Bacillus subtilis AraA), L-ribulokinase (Escherichia coli AraB) and L-ribulose-5-phosphate 4epimerase (E. coli AraD) [26,27], in combination with two different plasmid-borne xylose pathways, were constructed

  • Arabinose was instead converted to arabitol, which was believed to be catalysed by the overexpressed heterologous P. stipitis xylose reductase (XR) enzyme

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Summary

Introduction

Ethanolic fermentation of lignocellulosic biomass is a sustainable option for the production of bioethanol This process would greatly benefit from recombinant Saccharomyces cerevisiae strains able to ferment, besides the hexose sugar fraction, the pentose sugars, arabinose and xylose. Cost-effective and sustainable production of ethanol as a transportation fuel entails the utilisation of microbial strains able to ferment completely all the sugars in lignocellulosic hydrolyzates [4,5,6]. S. cerevisiae is unable to utilise arabinose and xylose, which in some raw materials such as agricultural residues and hardwoods, can account for more than 30% of total sugars [9], and which constitutes a significant barrier to the cost-effectiveness and sustainability of bioethanol production [6]. S. cerevisiae has been extensively engineered, developed and adapted to expand its substrate range to include the utilisation of the pentose sugars, arabinose and xylose, for growth and ethanol production [4]

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