•By-products of lignocellulose pretreatment inhibit microbial and enzymic biocatalysts.•Groups of inhibitors from components of lignocellulose are reviewed.•The review covers different strategies to ...alleviate inhibition problems.•Industrial implementation increases the relevance of inhibitor management.
Biochemical conversion of lignocellulosic feedstocks to advanced biofuels and other commodities through a sugar-platform process involves a pretreatment step enhancing the susceptibility of the cellulose to enzymatic hydrolysis. A side effect of pretreatment is formation of lignocellulose-derived by-products that inhibit microbial and enzymatic biocatalysts. This review provides an overview of the formation of inhibitory by-products from lignocellulosic feedstocks as a consequence of using different pretreatment methods and feedstocks as well as an overview of different strategies used to alleviate problems with inhibitors. As technologies for biorefining of lignocellulose become mature and are transferred from laboratory environments to industrial contexts, the importance of management of inhibition problems is envisaged to increase as issues that become increasingly relevant will include the possibility to use recalcitrant feedstocks, obtaining high product yields and high productivity, minimizing the charges of enzymes and microorganisms, and using high solids loadings to obtain high product titers.
By-products from hydrothermal pretreatment of lignocellulosic biomass inhibit enzymatic saccharification and microbial fermentation. Three long-chain organic extractants (Alamine 336, Aliquat 336 and ...Cyanex 921) were compared to two conventional organic solvents (ethyl acetate and xylene) with regard to conditioning of birch wood pretreatment liquid (BWPL) for improved fermentation and saccharification. In the fermentation experiments, extraction with Cyanex 921 resulted in the best ethanol yield, 0.34 ± 0.02 g g
−1
on initial fermentable sugars. Extraction with xylene also resulted in a relatively high yield, 0.29 ± 0.02 g g
−1
, while cultures consisting of untreated BWPL and BWPL treated with the other extractants exhibited no ethanol formation. Aliquat 336 was most efficient with regard to removing by-products, but the residual Aliquat after the extraction was toxic to yeast cells. Enzymatic digestibility increased by 19-33% after extraction with the long-chain organic extractants. The investigation demonstrates that conditioning with long-chain organic extractants has the potential to relieve inhibition of both enzymes and microbes.
Long-chain extractants with suitable properties improve both saccharification and fermentation.
•Effects of addition of gas on pretreatment under acidic conditions were explored.•Addition of O2 or N2 affected pretreatment severity and inhibitor formation.•The highest severity was observed for ...addition of O2.•Gas addition increased hemicellulose removal and overall glucan conversion.•Gas addition modulates the severity of acidic hydrothermal pretreatment.
The effects of the redox environment on acidic hydrothermal pretreatment were investigated in experiments with sugarcane bagasse (190 °C, 14 min) and Norway spruce (205 °C, 5 min). To modulate the redox environment, pretreatment was performed without gas addition, with N2, or with O2. Analyses covered pretreated solids, pretreatment liquids, condensates, enzymatic digestibility, and inhibitory effects of pretreatment liquids on yeast. Addition of gas, especially O2, resulted in increased severity, as reflected by up to 18 percent units lower recoveries of pretreated solids, up to 31 percent units lower glucan recoveries, improved hemicellulose removal, formation of pseudo-lignin, improved overall glucan conversion, and increased concentrations of several microbial inhibitors. Some inhibitors, such as formaldehyde and coniferyl aldehyde, did not, however, follow that pattern. TAC (Total Aromatic Content) values reflected inhibitory effects of pretreatment liquids. This study demonstrates how gas addition can be used to modulate the severity of acidic hydrothermal pretreatment.
•Steam pretreatment of spruce using H2SO4 and SO2 was compared at 195°C–215°C.•SO2 and high temperature caused partial degradation of cellulose and lignin.•SO2 and high temperature also promoted ...formation of particles of smaller size.•SO2 pretreatment liquid was more inhibitory to enzymic and microbial biocatalysts.•SO2 and high temperature resulted in superior enzymatic digestibility of cellulose.
Hydrothermal pretreatment improves bioconversion of lignocellulose, but the effects of different acid catalysts are poorly understood. The effects of sulfuric acid (SA) and sulfur dioxide (SD) in continuous steam pretreatment of wood of Norway spruce were compared in the temperature range 195°C–215°C. The inhibitory effects of the pretreatment liquid on cellulolytic enzymes and Saccharomyces cerevisiae yeast were higher for SD- than for SA-pretreated material, and the inhibitory effects increased with increasing pretreatment temperature. However, the susceptibility to cellulolytic enzymes of wood pretreated with SD was 2.0–2.9 times higher than that of wood pretreated with SA at the same temperature. Data conclusively show that the superior convertibility of SD-pretreated material was not due to inhibition phenomena but rather to the greater capability of the SD pretreatment to reduce the particle size through partial delignification and cellulose degradation. Particle size was shown to be correlated with enzymatic digestibility (R2 0.97–0.98).
Lignin derivatives have potential as antioxidants in advanced packaging materials through their ability to scavenge oxygen in reactions catalyzed by phenol-oxidizing enzymes, such as laccase. The ...effects of size fractionation of lignosulfonates on laccase-catalyzed reactions were investigated in experiments with aqueous solutions, films, and coated paperboard. Four industrial lignosulfonate preparations were compared:
(unfractionated),
(5-60 kDa enriched),
(≥60 kDa enriched), and
(≤60 kDa enriched). Extraction of lignosulfonates from films showed that the enzymic reaction increased the average molecular weight from <10,000 to up to 66,000. The enzymatic reaction resulted in an increase in the water contact angle of the films from the range 25-49° to 56-81°. The four preparations showed relatively small differences with regard to their ability to scavenge oxygen in aqueous solution and in experiments with coated paperboards in sealed chambers. Coatings with lignosulfonates where the contents of low-molecular weight material had been reduced (i.e.,
and
) showed improved water resistance after the enzymic reaction. Thus, in both aqueous and solid media, fractionation of lignosulfonates had little effect on oxygen scavenging, but fractionation was beneficial for other reasons, such as improved cross-linking resulting in higher molecular weight and superior water resistance.
Six lignocellulosic hydrolysates produced through acid pretreatment were analyzed for the occurrence of formaldehyde, acetaldehyde, and glycolaldehyde. Acetaldehyde was found in all six (0.3-1.6 mM) ...and formaldehyde in four (≤ 4.4 mM), whereas glycolaldehyde was not detected. To assess the relevance of these findings, fermentations with yeast and formaldehyde or acetaldehyde were performed in the concentration interval 0.5-10 mM. Formaldehyde already inhibited at 1.0 mM, whereas 5.0 mM acetaldehyde was needed to obtain a clear inhibitory effect. After 24 h of fermentation, 1.5 mM formaldehyde reduced the glucose consumption by 85%, the balanced ethanol yield by 92%, and the volumetric productivity by 91%. The results show that formaldehyde and acetaldehyde are prevalent in pretreated lignocellulose and that formaldehyde in some cases could explain a large part of the inhibitory effects on yeast by lignocellulosic hydrolysates, as three of six hydrolysates contained ≥ 1.9 mM formaldehyde, which was shown to be strongly inhibitory.
The enzymatic hydrolysis of cellulose is inhibited by non-productive adsorption of cellulases to lignin, and that is particularly problematic with lignin-rich materials such as softwood. Although ...conventional surfactants alleviate non-productive adsorption, using biosurfactants in softwood hydrolysis has not been reported. In this study, the effects of four biosurfactants, namely horse-chestnut escin,
rhamnolipid, and saponins from red and white quinoa varieties, on the enzymatic saccharification of steam-pretreated spruce were investigated. The used biosurfactants improved hydrolysis, and the best-performing one was escin, which led to cellulose conversions above 90%, decreased by around two-thirds lignin inhibition of Avicel hydrolysis, and improved hydrolysis of pretreated spruce by 24%. Red quinoa saponins (RQS) addition resulted in cellulose conversions above 80%, which was around 16% higher than without biosurfactants, and it was more effective than adding rhamnolipid or white quinoa saponins. Cellulose conversion improved with the increase in RQS addition up to 6 g/100 g biomass, but no significant changes were observed above that dosage. Although saponins are known to inhibit yeast growth, no inhibition of
fermentation of hydrolysates produced with RQS addition was detected. This study shows the potential of biosurfactants for enhancing the enzymatic hydrolysis of steam-pretreated softwood.
Display omitted
•Biomass of Nordic microalgae was used as feed for bacterial biopolymer production.•Thermochemical pre-treatment with 1% H2SO4 improved the solubilization of total sugars.•Maximal PHB ...content (27% of DW) was achieved with hydrolysate of Chlorococcum sp.•Nordic microalgae can combine wastewater reclamation with PHB production.
Biomass from four different Nordic microalgal species, grown in BG-11 medium or synthetic wastewater (SWW), was explored as inexpensive carbohydrate-rich feedstock for polyhydroxybutyrate (PHB) production via microbial fermentation. Thermochemical pre-treatment (acid treatment followed by autoclavation) with 2% hydrochloric acid or 1% sulphuric acid (v/v) was used to maximize sugar yield prior to fermentation. Pre-treatment resulted in ∼5-fold higher sugar yield compared to the control. The sugar-rich hydrolysate was used as carbon source for the PHB-producing extremophilic bacterium Halomonas halophila. Maximal PHB production was achieved with hydrolysate of Chlorococcum sp. (MC-1) grown on BG-11 medium (0.27 ± 0.05 g PHB/ g DW), followed by hydrolysate derived from Desmodesmus sp. (RUC-2) grown on SWW (0.24 ± 0.05 g PHB/ g DW). Nordic microalgal biomass grown on wastewater therefore can be used as cheap feedstock for sustainable bioplastic production. This research highlights the potential of Nordic microalgae to develop a biobased economy.
•Microbial inhibitors in steam-pretreated softwood were characterized.•Effects of impregnating agents and temperature on inhibitor formation were compared.•Storage of pretreated biomass affects the ...concentrations of some inhibitors.•Among the inhibitors, the toxic effects of formaldehyde were the most significant.
Wood chips of Norway spruce were pretreated by steam explosion at 195–215 °C after impregnation with either sulfuric acid (SA) or sulfur dioxide (SD). The effects of different pretreatment conditions on formation of microbial inhibitors were investigated, and the inhibitory effects on yeast of pretreatment liquids and of specific inhibitors that were found in the pretreatment liquids were elucidated. Whereas the concentrations of most inhibitors increased with increasing pretreatment temperatures, there were exceptions, such as formaldehyde and p-hydroxybenzaldehyde. The highest concentration of each inhibitor was typically found in SD-pretreated material, but formic acid was an exception. The toxic effects on yeast were studied using concentrations corresponding to loadings of 12 and 20% total solids (TS). Among individual inhibitors that were quantitated in pretreatment liquids, the concentrations of formaldehyde were by far most toxic. There was no or minimal yeast growth in the formaldehyde concentration range (5.8–7.7 mM) corresponding to 12% TS.
PDF
