Continuous Enzymatic Hydrolysis Development for Improved Saccharification Performance

Abstract

The Continuous Enzymatic Hydrolysis Development (CEHD) project aims to reduce the cost and commercialization risks of Gen2 biorefinery sugar/lignin/ethanol production through development of a deployable continuous enzymatic hydrolysis process. Recent changes in the technical landscape of commercial enzymatic hydrolysis of Gen2 pretreated biomass dictate that the existing hybrid SSF approach be reconsidered. Most importantly, the current practice of "finishing hydrolysis" in SSF must be abandoned due to the fact that new cellulase/hemicellulose formulations from Novozymes, now the sole supplier of commercial Gen2 enzymes in North America, are now not rated for SSF (see NZ CTec3HS product bulletin). We have recently developed bench scale CEH tools to optimize saccharification of DMR pretreated biomass where, unlike SSF with yeast or Zymomonas, the pH, temperature, oxygen tension, LPMO mediator concentration, and/or removal of end-product inhibitors can be precisely controlled. In scale up, the goal is to use existing commercial cross flow ceramic membrane filtration external loops coupled to enzymatic hydrolysis (EH) reactors. Pretreated biomass solids and enzymes are retained for reaction while solubilized product sugars are removed in situ, with high extents of conversion and longer enzyme lifetimes achieved through a series of reactor-membrane unit stages. The CEHD project is focused on advancing CEH as a transformational, process-intensified, lower-cost method for producing soluble clarified biomass sugars and insoluble lignin-rich streams.