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Botany JoVE (Open Access) Citable · DOI

High-throughput Saccharification Assay for Lignocellulosic Materials

DOI: 10.3791/3240-v
What you'll learn
  • ✓ Prepare plant biomass samples in 96-well plate format for high-throughput analysis
  • ✓ Perform pretreatment and enzymatic hydrolysis on lignocellulosic materials
  • ✓ Quantify reducing sugars released and assess saccharification potential
Protocol

A simple, rapid method for determining the saccharification potential of large numbers of plant biomass samples is described. The automated platform for this analysis involves the preparation of the plant biomass for analysis in 96 well plates and the subsequent performance of pretreatment, hydrolysis and quantification of the sugars released.

Difficulty
intermediate
Total time
~4–6 hours per batch (96 samples); overnight incubation recommended
Biosafety
BSL-1

Steps

1
Prepare plant biomass samples in 96-well plates

Dispense and organize lignocellulosic plant biomass samples into 96-well plate format for automated processing. This enables parallel analysis of multiple samples simultaneously.

▶ 05:03
2
Perform pretreatment of biomass samples

Apply pretreatment procedure to plant biomass in 96-well plates to increase accessibility of cellulose and hemicellulose for enzymatic breakdown.

▶ 05:03
3
Execute enzymatic hydrolysis reaction

Incubate pretreated biomass with cellulase enzymes to degrade cellulose and hemicellulose into fermentable sugars.

▶ 06:40
4
Quantify reducing sugars from hydrolysis

Measure total reducing sugars released using colorimetric or enzymatic detection methods to determine saccharification efficiency and potential of each sample.

▶ 07:45
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