title: Methodology for enabling high-throughput simultaneous saccharification and fermentation screening of yeast using solid biomass as a substrate.
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 hydrolyzed 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 utilizing sheets of biomass of consistent density (handbills), for paper, and slurries of pretreated biomass that could be pipetted allowed standardized and accurate transfers to 96-well plates to 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 trialed successfully for SSFs at 1 mL volume. The methodology was successfully tested by showing the effects of steam explosion pretreatment on both oilseed 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 standardized assay techniques such as simple colorimetric, High-performance liquid chromatography (HPLC) and Nuclear magnetic resonance (NMR) spectroscopy. Furthermore this research has practical uses in the biorefining of biomass substrates for second generation biofuels and novel biobased chemicals by allowing HTP SSF screening, which should allow selected samples to be scaled up or studied in more detail.
authors: The Biorefinery Centre, Institute of Food Research, Norwich Research Park, Colney, Norwich, NR4 7UA UK.
authors: Adam
authors: A
authors: Elliston
authors: The Biorefinery Centre, Institute of Food Research, Norwich Research Park, Colney, Norwich, NR4 7UA UK.
authors: Ian P
authors: IP
authors: Wood
authors: The Biorefinery Centre, Institute of Food Research, Norwich Research Park, Colney, Norwich, NR4 7UA UK ; École supérieure d'ingénieurs Réunion Océan Indien, Génie Biologique, Université de La Réunion, Parc Technologique Universitaire, 2 Rue Joseph Wetzell, 97490 Sainte-Clotilde, La Réunion France.
authors: Marie J
authors: MJ
authors: Soucouri
authors: The Biorefinery Centre, Institute of Food Research, Norwich Research Park, Colney, Norwich, NR4 7UA UK ; Institut Universitaire de Technologie, Universite de la Reunion, 40 avenue de Soweto, BP 373, 97455 Saint-Pierre Cedex, La Réunion France.
authors: Rachelle J
authors: RJ
authors: Tantale
authors: The National Collection of Yeast Cultures, Institute of Food Research, Norwich Research Park, Colney, Norwich, NR4 7UA UK.
authors: Jo
authors: J
authors: Dicks
authors: The National Collection of Yeast Cultures, Institute of Food Research, Norwich Research Park, Colney, Norwich, NR4 7UA UK.
authors: Ian N
authors: IN
authors: Roberts
authors: The Biorefinery Centre, Institute of Food Research, Norwich Research Park, Colney, Norwich, NR4 7UA UK.
authors: Keith W
authors: KW
authors: Waldron
keywords: Biomass; Biorefining; Cellulosic biomass; Cellulosic ethanol; Enzyme saccharification; High-throughput screening; Simultaneous saccharification and fermentation
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