Harnessing the potential of LPMO-containing cellulase cocktails poses new demands on processing conditions.

Vincent G H Eijsink,Gerdt Müller,Anikó Várnai,Svein Jarle Horn,Katja Salomon Johansen

doi:10.1186/s13068-015-0376-y

Abstract

BackgroundThe emerging bioeconomy depends on improved methods for processing of lignocellulosic biomass to fuels and chemicals. Saccharification of lignocellulose to fermentable sugars is a key step in this regard where enzymatic catalysis plays an important role and is a major cost driver. Traditionally, enzyme cocktails for the conversion of cellulose to fermentable sugars mainly consisted of hydrolytic cellulases. However, the recent discovery of lytic polysaccharide monooxygenases (LPMOs), which cleave cellulose using molecular oxygen and an electron donor, has provided new tools for biomass saccharification.ResultsCurrent commercial enzyme cocktails contain LPMOs, which, considering the unique properties of these enzymes, may change optimal processing conditions. Here, we show that such modern cellulase cocktails release up to 60 % more glucose from a pretreated lignocellulosic substrate under aerobic conditions compared to anaerobic conditions. This higher yield correlates with the accumulation of oxidized products, which is a signature of LPMO activity. Spiking traditional cellulase cocktails with LPMOs led to increased saccharification yields, but only under aerobic conditions. LPMO activity on pure cellulose depended on the addition of an external electron donor, whereas this was not required for LPMO activity on lignocellulose.ConclusionsIn this study, we demonstrate a direct correlation between saccharification yield and LPMO activity of commercial enzyme cocktails. Importantly, we show that the LPMO contribution to overall efficiency may be large if process conditions are adapted to the key determinants of LPMO activity, namely the presence of electron donors and molecular oxygen. Thus, the advent of LPMOs has a great potential, but requires rethinking of industrial bioprocessing procedures.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-015-0376-y) contains supplementary material, which is available to authorized users.

Highlights

The emerging bioeconomy depends on improved methods for processing of lignocellulosic biomass to fuels and chemicals
Results it is well established that molecular oxygen and an electron donor are required for the action of lytic polysaccharide monooxygenase (LPMO) [2], still little is known about the impact of these requirements on the efficiency of commercial cellulase mixtures [18, 19]
The results below show that Glc4gemGlc accumulated during saccharification of lignocellulosic materials involving LPMO activity, indicating that this dimeric product cannot be cleaved by the β-glucosidase activity in the enzyme cocktail

Summary

Introduction

The emerging bioeconomy depends on improved methods for processing of lignocellulosic biomass to fuels and chemicals. Saccharification of lignocellulose to fermentable sugars is a key step in this regard where enzymatic catalysis plays an important role and is a major cost driver. The recent discovery of lytic polysaccharide monooxygenases (LPMOs), which cleave cellulose using molecular oxygen and an electron donor, has provided new tools for biomass saccharification. The high efficiency of the most recently developed commercial enzyme cocktails has been an important factor in making cellulosic ethanol a commercial reality [10]. LPMO activity depends on the presence of oxygen [2], which is likely to be an important factor to consider when designing industrial saccharification and fermentation processes

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Discussion

Conclusion