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High-Throughput Screening Techniques for Biomass Conversion

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Abstract

High-throughput (HTP) screening of biomass or biomass-degrading enzymes, regardless of the desired outcome, is fraught with obstacles and challenges not typically faced in more traditional biotechnology. The enzyme systems are complex and synergistic and the substrate is highly heterogeneous, insoluble, and difficult to dispense. Digestions are often carried out for days at temperatures of 50°C or higher, leading to significant challenges regarding evaporation control in small well volumes. Furthermore, it is often desirable to condition or “pretreat” the biomass at extreme temperatures and/or pH to enhance enzyme digestibility. Once the substrate has been saccharified, evaluation of the extent and efficiency of conversion is made more difficult by time-consuming and tedious techniques used to measure the sugar products. Over the past decade or so, biomass researchers have creatively addressed these challenges by developing techniques to reduce biomass heterogeneity, uniformly distribute biomass samples at the small scale, pretreat the biomass at the small scale, quantitatively load these samples with enzymes, control evaporation of small reaction volumes for multiday incubations, and rapidly quantify the products. Other aspects of these measurements remain problematic and are being addressed. This review will address some of these challenges in detail, but more importantly, we will endeavor to educate the reader about the trials, tribulations, and pitfalls of carrying out HTP screening in biomass conversion research.

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Abbreviations

BESC:

BioEnergy Science Center

NREL:

National Renewable Energy Laboratory

HTP:

high throughput

US DOE:

United States Department of Energy

MS:

mass spectrometry

GC:

gas chromatography

HPLC:

high-performance liquid chromatography

UPLC:

ultra-performance liquid chromatography

SBS:

Society for Biomolecular Screening

AFEX:

ammonium fiber expansion

DNS:

dinitrosalicylic Acid

BCA:

bicinchoninic Acid

MBTH:

3-methyl-2-benzothiazolinonehydrazone

RI:

refractive index

PAD:

pulsed amperometric detection

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Acknowledgments

The authors wish to thank Charles Wyman and his coworkers at the University of California-Riverside for the description and images of their high-throughput pretreatment reactor system. This work was supported by the DOE Office of Science, Office of Biological and Environmental Research through the BioEnergy Science Center (BESC), a DOE Bioenergy Research Center.

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Authors and Affiliations

  1. Biosciences Center, National Renewable Energy Laboratory, 1617 Cole Boulevard, MS 3323, Golden, CO, 80401, USA

    Stephen R. Decker, Roman Brunecky, Michael E. Himmel & Michael J. Selig

  2. National Bioenergy Center, National Renewable Energy Laboratory, Golden, CO, USA

    Melvin P. Tucker

Authors
  1. Stephen R. Decker
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  2. Roman Brunecky
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  3. Melvin P. Tucker
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  4. Michael E. Himmel
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  5. Michael J. Selig
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Correspondence to Stephen R. Decker.

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Decker, S.R., Brunecky, R., Tucker, M.P. et al. High-Throughput Screening Techniques for Biomass Conversion. Bioenerg. Res. 2, 179–192 (2009). https://doi.org/10.1007/s12155-009-9051-0

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