Abstract

BackgroundSugarcane bagasse is a major source of lignocellulosic biomass, yet its economic potential is not fully realised. To add value to bagasse, processing is needed to gain access to the embodied recalcitrant biomaterials. When bagasse is stored in piles in the open for long periods it is colonised by microbes originating from the sugarcane, the soil nearby or spores in the environment. For these microorganisms to proliferate they must digest the bagasse to access carbon for growth. The microbial community in bagasse piles is thus a potential resource for the discovery of useful and novel microbes and industrial enzymes. We used culturing and metabarcoding to understand the diversity of microorganisms found in a uniquely undisturbed bagasse storage pile and screened the cultured organisms for fibre-degrading enzymes.ResultsSamples collected from 60 to 80 cm deep in the bagasse pile showed hemicellulose and partial lignin degradation. One hundred and four microbes were cultured from different layers and included a high proportion of oleaginous yeast and biomass-degrading fungi. Overall, 70, 67, 70 and 57% of the microbes showed carboxy-methyl cellulase, xylanase, laccase and peroxidase activity, respectively. These percentages were higher in microbes selectively cultured from deep layers, with all four activities found for 44% of these organisms. Culturing and amplicon sequencing showed that there was less diversity and therefore more selection in the deeper layers, which were dominated by thermophiles and acid tolerant organisms, compared with the top of pile. Amplicon sequencing indicated that novel fungi were present in the pile.ConclusionsA combination of culture-dependent and independent methods was successful in exploring the diversity in the bagasse pile. The variety of species that was found and that are known for biomass degradation shows that the bagasse pile was a valuable selective environment for the identification of new microbes and enzymes with biotechnological potential. In particular, lignin-modifying activities have not been reported previously for many of the species that were identified, suggesting future studies are warranted.

Highlights

  • Sugarcane bagasse is a major source of lignocellulosic biomass, yet its economic potential is not fully realised

  • Oil-producing yeast, biomass-degrading fungi, Bacillus and Streptomyces cultured from the bagasse In total, 104 microbes were cultured from bagasse samples collected at the Rocky Point sugarcane mill in May 2016 and February 2017

  • The strains and how they were selected are summarised in Additional file 1. 16S or Internal transcriber sequence (ITS) sequences were used to query the 16S ribosomal sequence database at The National Centre for Biotechnology Information (NCBI) or the UNITE [35] database, respectively

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Summary

Introduction

Sugarcane bagasse is a major source of lignocellulosic biomass, yet its economic potential is not fully realised. At most mills the bagasse is used to fuel boilers, cogenerating steam and electricity [2] It could have various higher value uses: second generation biofuels [3]; fibres for paper, particle board [4] and in 3D printing [5]; xylan-based products such as xylooligosaccharides [6]; substrate for edible or medicinal mushroom growth [7]; substrate for single-cell protein, enzyme, or other highvalue microbial products [8, 9]; and high value chemicals from the lignin fraction [10] are some examples. Biological processing is one approach and progress in our understanding of biomass decomposition is continually improving, there is still potential to further exploit the genetic diversity of adapted microbial communities inhabiting environments rich in lignocellulose

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