Advertisement

Engineering of a high-throughput screening system to identify cellulosic biomass, pretreatments, and enzyme formulations that enhance sugar release

Authors

  • Michael H. Studer,

    1. Chemical and Environmental Engineering Department, Center for Environmental Research and Technology, Bourns College of Engineering, University of California Riverside, 1084 Columbia Avenue, Riverside, California 92507; telephone: +951-781-5791; fax: +951-781-5790
    Search for more papers by this author
  • Jaclyn D. DeMartini,

    1. Chemical and Environmental Engineering Department, Center for Environmental Research and Technology, Bourns College of Engineering, University of California Riverside, 1084 Columbia Avenue, Riverside, California 92507; telephone: +951-781-5791; fax: +951-781-5790
    Search for more papers by this author
  • Simone Brethauer,

    1. Chemical and Environmental Engineering Department, Center for Environmental Research and Technology, Bourns College of Engineering, University of California Riverside, 1084 Columbia Avenue, Riverside, California 92507; telephone: +951-781-5791; fax: +951-781-5790
    Search for more papers by this author
  • Heather L. McKenzie,

    1. Chemical and Environmental Engineering Department, Center for Environmental Research and Technology, Bourns College of Engineering, University of California Riverside, 1084 Columbia Avenue, Riverside, California 92507; telephone: +951-781-5791; fax: +951-781-5790
    Search for more papers by this author
  • Charles E. Wyman

    Corresponding author
    1. Chemical and Environmental Engineering Department, Center for Environmental Research and Technology, Bourns College of Engineering, University of California Riverside, 1084 Columbia Avenue, Riverside, California 92507; telephone: +951-781-5791; fax: +951-781-5790
    • Chemical and Environmental Engineering Department, Center for Environmental Research and Technology, Bourns College of Engineering, University of California Riverside, 1084 Columbia Avenue, Riverside, California 92507; telephone: +951-781-5791; fax: +951-781-5790.
    Search for more papers by this author

Abstract

The recalcitrance of cellulosic biomass, the only abundant, sustainable feedstock for making liquid fuels, is a primary obstacle to low cost biological processing, and development of more easily converted plants and more effective enzymes would be of great benefit. Because no single parameter describes recalcitrance, superior variants can only be identified by measuring sugar release from plants subjected to pretreatment and enzymatic hydrolysis. However, genetic modifications of plants coupled with molecular engineering of deconstruction proteins and definition of pretreatment conditions create a very large sample set, and previous methods for biomass pretreatment at elevated temperatures and pressures prevented use of a fully integrated high-throughput (HTP) screening pipeline. Herein, we report on the engineering of a novel HTP pretreatment system employing a 96 well-plate format that withstands extreme pretreatment conditions for rapid screening of biomass–enzyme-pretreatment combinations. This includes the development of new approaches to steam heating and water quenching the system that result in much faster heat up and cool down than previously possible and show consistent temperature histories across the multiwell plate. Coupled pretreatment and enzymatic hydrolysis performance of the well plate pretreatment system is shown to be consistent among the many wells in the device and also with performance of conventional tubular reactors. Biotechnol. Bioeng. 2010; 105: 231–238. © 2009 Wiley Periodicals, Inc.

Ancillary