Methodology for enabling high-throughput simultaneous saccharification and fermentation (SSF) screening of yeast using solid biomass as a substrate

Elliston A., Wood I. P., Soucouri M. J., Tantale R. J., Dicks J., Roberts I. N., Waldron K. W.. (2015)

Biotechnology for biofuels, 8, 2

Abstract Background High-throughput (HTP) screening is becoming an increasingly useful tool for collating biological data which would otherwise require the employment of excessive resources. Second generation biofuel production is one such process. HTP screening allows the investigation of large sample sets to be undertaken with increased speed and cost effectiveness. This paper outlines a methodology that will enable solid lignocellulosic substrates to be hydrolysed and fermented at a 96 well plate scale, facilitating HTP screening of ethanol production, whilst maintaining repeatability similar to that achieved at a larger scale. Results The results showed that utilising sheets of biomass of consistent density (handbills), for paper, and pipettable slurries of pretreated biomass allowed standardised and accurate transfers to 96 well plates be achieved (±3.1 and 1.7% respectively).  Processing these substrates by simultaneous saccharification and fermentation (SSF) at various volumes showed no significant difference on final ethanol yields; either at standard shake flask (200 mL), universal bottle (10 mL) or 96 well plate (1 mL) scales. Substrate concentrations of up to 10% (w/v) were trialled successfully for SSFs at 1 mL volume. The methodology was successfully tested by showing the effects of steam explosion pretreatment on both oil seed rape and wheat straws.     Conclusions This methodology could be used to replace large shake flask reactions with comparatively fast 96 well plate SSF assays allowing for HTP experimentation. Additionally this method is compatible with a number of standardised assay techniques such as simple colorimetric, HPLC and NMR. Furthermore this research has practical uses in biorefining of biomass substrates for second generation biofuels and novel bio-based chemicals by allowing high-throughput simultaneous saccharification and fermentation screening which should allow selected samples to be scaled up or studied in more detail.

Biotechnology for biofuels, 8, 2

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