Abstract

BackgroundDeveloping novel microbial cell factories requires careful testing of candidates under industrially relevant conditions. However, this frequently occurs late during the strain development process. The availability of laboratory media that simulate industrial-like conditions might improve cell factory development, as they allow for strain construction and testing in the laboratory under more relevant conditions. While sugarcane molasses is one of the most important substrates for the production of biofuels and other bioprocess-based commodities, there are no defined media that faithfully simulate it. In this study, we tested the performance of a new synthetic medium simulating sugarcane molasses.ResultsLaboratory scale simulations of the Brazilian ethanol production process, using both sugarcane molasses and our synthetic molasses (SM), demonstrated good reproducibility of the fermentation performance, using yeast strains, PE-2 and Ethanol Red™. After 4 cycles of fermentation, the final ethanol yield (gp gs−1) values for the SM ranged from 0.43 ± 0.01 to 0.44 ± 0.01 and from 0.40 ± 0.01 to 0.46 ± 0.01 for the molasses-based fermentations. The other fermentation parameters (i.e., biomass production, yeast viability, and glycerol and acetic acid yield) were also within similar value ranges for all the fermentations. Sequential pairwise competition experiments, comparing industrial and laboratory yeast strains, demonstrated the impact of the media on strain fitness. After two sequential cocultivations, the relative abundance of the laboratory yeast strain was 5-fold lower in the SM compared to the yeast extract-peptone-dextrose medium, highlighting the importance of the media composition on strain fitness.ConclusionsSimulating industrial conditions at laboratory scale is a key part of the efficient development of novel microbial cell factories. In this study, we have developed a synthetic medium that simulated industrial sugarcane molasses media. We found good agreement between the synthetic medium and the industrial media in terms of the physiological parameters of the industrial-like fermentations.

Highlights

  • Developing novel microbial cell factories requires careful testing of candidates under industrially relevant conditions

  • To develop the synthetic molasses (SM) medium, a basal molasses medium, described elsewhere [29], was modified based on the average sugarcane molasses composition described in other studies [10, 26, 34, 40,41,42], in order to obtain a final chemical composition similar to those observed in actual sugarcane molasses samples

  • An overview of all SM formulations developed throughout this study, and reasons for altering them are depicted in Additional file 1

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Summary

Introduction

Developing novel microbial cell factories requires careful testing of candidates under industrially relevant conditions. This frequently occurs late during the strain development process. The availability of laboratory media that simulate industrial-like conditions might improve cell factory development, as they allow for strain construction and testing in the laboratory under more relevant conditions. While sugarcane molasses is one of the most important substrates for the production of biofuels and other bioprocess-based commodities, there are no defined media that faithfully simulate it. Molasses is produced via the separation of sucrose crystals that follow the water evaporation from clarified juice (from sugarcane or beet) during the production of crystal sugar. Molasses can be further recycled in this process in order to maximize sugar production. As a rule of thumb, the higher the number of recycling steps the molasses is subjected to, the poorer is its quality as a raw material for fermentation [10]

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