Microplate-Based Evaluation of the Sugar Yield from Giant Reed, Giant Miscanthus and Switchgrass after Mild Chemical Pre-Treatments and Hydrolysis with Tailored Trichoderma Enzymatic Blends

Appl Biochem Biotechnol. 2017 Nov;183(3):876-892. doi: 10.1007/s12010-017-2470-z. Epub 2017 Apr 6.

Abstract

Giant reed, miscanthus, and switchgrass are considered prominent lignocellulosic feedstocks to obtain fermentable sugars for biofuel production. The bioconversion into sugars requires a delignifying pre-treatment step followed by hydrolysis with cellulase and other accessory enzymes like xylanase, especially in the case of alkali pre-treatments, which retain the hemicellulose fraction. Blends richer in accessory enzymes than commercial mix can be obtained growing fungi on feedstock-based substrates, thus ten selected Trichoderma isolates, including the hypercellulolytic strain Trichoderma reesei Rut-C30, were grown on giant reed, miscanthus, or switchgrass-based substrates. The produced enzymes were used to saccharify the corresponding feedstocks, compared to a commercial enzymatic mix (6 FPU/g). Feedstocks were acid (H2SO4 0.2-2%, w/v) or alkali (NaOH 0.02-0.2%, w/v) pre-treated. A microplate-based approach was chosen for most of the experimental steps due to the large number of samples. The highest bioconversion was generally obtained with Trichoderma harzianum Or4/99 enzymes (78, 89, and 94% final sugar yields at 48 h for giant reed, miscanthus, and switchgrass, respectively), with significant increases compared to the commercial mix, especially with alkaline pre-treatments. The differences in bioconversion yields were only partially caused by xylanases (maximum R 2 = 0.5), indicating a role for other accessory enzymes.

Keywords: Acid pre-treatment; Alkaline pre-treatment; Arundo donax; Cellulase; Lignocellulose; Miscanthus × giganteus; Panicum virgatum; Reducing sugars; Trichoderma; Xylanase.

MeSH terms

  • Biotechnology / instrumentation
  • Biotechnology / methods*
  • Carbohydrate Metabolism*
  • Cellulase / metabolism*
  • Endo-1,4-beta Xylanases / metabolism*
  • Hydrolysis
  • Lignin / metabolism
  • Microtechnology / instrumentation
  • Microtechnology / methods*
  • Panicum / metabolism*
  • Trichoderma / enzymology*

Substances

  • lignocellulose
  • Lignin
  • Cellulase
  • Endo-1,4-beta Xylanases