Acid Pretreated Substrates Research Articles

Effects of different pelleting technologies and parameters on pretreatment and enzymatic saccharification of lignocellulosic biomass

Densification of lignocellulosic biomass is beneficial for its logistics, and in some situations, also for its application in biorefineries. In this study, industrial pellets and laboratory-made pellets produced at different die temperatures (90, 125 °C) from wheat straw, beech, and pine were pretreated (by dilute acid and alkali) and enzymatically hydrolyzed to study the effects of pelleting on pretreatment and sugar yield. The results showed that industrial pelleting positively affected sugar yield of the three biomasses for both acid and alkaline pretreatments, while laboratory pelleting affected sugar yield differently related to pretreatment types and biomass species. Pelleting disturbed biomass cell wall structure and affected the stability of hemicellulose and β-O-4′ linkages in lignin. The extent of hemicellulose solubility during acid pretreatment and the amount of lignin β-O-4′ ether linkages present in acid pretreated substrates could serve as indicators of the effect of pelleting on sugar yield. The structural modifications caused by pelleting and their effects on the enzymatic sugar yield correlate tightly to pelleting technology, pretreatment method, and biomass species. The discrepant effects between industrial and laboratory pelleting on sugar yield observed in this study should be considered when designing future studies.

The Effect of Acid Pickling on the Corrosion Behavior of a Cerium Conversion-Coated AA2198-T851 Al-Cu-Li Alloy

The effect of acid pickling pretreatments prior to cerium conversion coating process on the corrosion behavior of AA2198-T851 alloy substrates was investigated. Three acid pretreatments were employed: nitric acid (HNO3), phosphoric acid (H3PO4) and sulfuric acid (H2SO4). The cerium conversion coating process was performed using a batch solution composed of cerium nitrate and hydrogen peroxide. Microscopic techniques, electrochemical impedance spectroscopy, polarization resistance and open-circuit potential measurements were employed to investigate the effect of each acid pretreatment. The untreated and nitric acid pretreated substrates presented more defective cerium conversion layers than the substrates treated with phosphoric and sulfuric acids. Accordingly, the corrosion resistance of the untreated and nitric acid-treated substrates was very low, while that of the substrates treated with phosphoric acid and sulfuric acids were greatly improved. The sulfuric acid pickling treatment was the best pretreatment before cerium conversion coating among the investigated pretreatments on the AA2198-T851 Al-Cu-Li alloy.

Evaluation of the action of Tween 20 non-ionic surfactant during enzymatic hydrolysis of lignocellulose: Pretreatment, hydrolysis conditions and lignin structure

The aim of this work was to study the effects of pretreatment process, hydrolysis condition and structural features of lignin on the improving action of surfactants (Tween 20) for enzymatic hydrolysis of pretreated wheat straw, and further to interpret the relation of these factors with the non-productive adsorption of cellulases on lignin. Tween 20 seemed to be more greatly improve cellulose conversion under harsher conditions. The surfactant showed more significant improvement for acid-pretreated substrates than oxidative-pretreated substrates. Highly-condensed lignin and phenolic hydroxyl groups showed much stronger adsorption ability to cellulases, while Tween 20 could well block the lignin–cellulase interactions recovering cellulose hydrolyzability. It was proposed that pretreatments altered lignin structures, resulting in the change of surface properties thus further impacting the lignin–cellulase interactions. Addition of Tween 20 could modify lignin surface properties to change its hydrophobicity, hydrogen bonding ability and surface charges, thus reducing the non-productive adsorption of proteins.

Enhanced enzymatic digestibility of mixed wood sawdust by lignin modification with naphthol derivatives during dilute acid pretreatment

Effects of the addition of 2-naphthol and 2-naphthol-7-sulfonate on the dilute acid pretreatment of mixed wood sawdust were investigated, respectively. Compared to 2-naphthol, 2-naphtnol-7-sulfonate was more effective to enhance delignification and facilitate the enzymatic hydrolysis. The 72 h hydrolysis yield was improved by 47.8% for 2-naphthol-7-sulfone, while only 9.1% was observed for 2-naphthol. The surface charges, enzyme adsorption, and cellulose accessibility of dilute acid pretreated substrates with or without naphthol derivatives were examined. The improved enzymatic hydrolysis by adding 2-naphthol-7-sulfonate was ascribed to the higher negative surface charges, the lower enzyme non-productive binding, and the higher cellulose accessibility of pretreated substrates. Additionally, the HSQC NMR and 31P NMR analysis were carried out on both decomposed lignins and residual bulk lignins. It indicated that the addition of the naphthol derivatives during pretreatment could suppress the lignin repolymerization, which further mitigated the inhibition of residual lignins on enzymatic hydrolysis.

Integrated chemical and multi-scale structural analyses for the processes of acid pretreatment and enzymatic hydrolysis of corn stover

Corn stover was pretreated with acid under moderate conditions (1.5%, w/w, 121°C, 60min), and kinetic enzymolysis experiments were performed on the pretreated substrate using a mixture of Celluclast 1.5L (20FPU/g dry substrate) and Novozyme 188 (40CBU/g dry substrate). Integrated chemical and multi-scale structural methods were then used to characterize both processes. Chemical analysis showed that acid pretreatment removed considerable hemicellulose (from 19.7% in native substrate to 9.28% in acid-pretreated substrate) and achieved a reasonably high conversion efficiency (58.63% of glucose yield) in the subsequent enzymatic hydrolysis. Multi-scale structural analysis indicated that acid pretreatment caused structural changes via cleaving acetyl linkages, solubilizing hemicellulose, relocating cell wall surfaces and enlarging substrate porosity (pore volume increased from 0.0067cm3/g in native substrate to 0.019cm3/g in acid-pretreated substrate), thereby improving the polysaccharide digestibility.

Non-ionic surfactants do not consistently improve the enzymatic hydrolysis of pure cellulose

Non-ionic surfactants have been frequently reported to improve the enzymatic hydrolysis of pretreated lignocellulosic biomass and pure cellulose. However, how the hydrolysis condition, substrate structure and cellulase formulation affect the beneficial action of surfactants has not been well elucidated. In this work, it was found that the enzymatic hydrolysis of pure cellulose was not consistently improved by surfactants. Contrarily, high surfactant concentration, e.g. 5g/L, which greatly improved the hydrolysis of dilute acid pretreated substrates, actually showed notable inhibition to pure cellulose conversion in the late phase of hydrolysis. Under an optimal hydrolysis condition, the improvement by surfactant was limited, but under harsh conditions surfactant indeed could enhance cellulose conversion. It was proposed that non-ionic surfactants could interact with substrates and cellulases to impact the adsorption behaviors of cellulases. Therefore, the beneficial action of surfactants on pure cellulose hydrolysis is influenced by hydrolysis condition, cellulose structural features and cellulase formulation.

Effect of pretreatments and endo-1,4-β-xylanase hydrolysis of canola meal and mustard bran for production of oligosaccharides.

Alkali/acid-pretreated canola meal and mustard bran were subjected to endo-1,4-β-xylanase (T. longibrachiatum) hydrolysis for oligosaccharide production. Pretreatments significantly (α = 0.05) increased the relative content of pentose sugars, especially in alkali-pretreated canola meal (∼44 %) and mustard bran (∼72 %). The amounts of pentosan (g/100 g) in acid- and alkali-pretreated canola meal were 7.50 and 8.21 and in corresponding mustard bran were 8.67 and 10.39, respectively. These pretreated substrates produced a pentose content (g/100 g) of 2.10 ± 0.14 (18 h) and 2.95 ± 0.10 (24 h), respectively, during hydrolysis. As per UPLC-MS data, the main oligosaccharides in the hydrolyzates of alkali-pretreated substrates are xylo-glucuronic acid and xylobiose. The release of total phenolics of the hydrolyzates increased until 18 h irrespective of the type of substrate or pretreatment. Hydrolyzates of acid-pretreated substrates indicated more total antioxidant activity than alkali-pretreated substrates, attributed to its high phenolic content. The study suggests the potential of canola meal and mustard bran for the production of oligosaccharides, wherein the use of various combinations of cell-wall-degrading enzymes and its optimization may result in a better yield, with simultaneous production of endogenous phenolics.

Effects of SPORL and Dilute Acid Pretreatment on Substrate Morphology, Cell Physical and Chemical Wall Structures, and Subsequent Enzymatic Hydrolysis of Lodgepole Pine

The effects of pretreatment by dilute acid and sulfite pretreatment to overcome recalcitrance of lignocellulose (SPORL) on substrate morphology, cell wall physical and chemical structures, along with the subsequent enzymatic hydrolysis of lodgepole pine substrate were investigated. FE-SEM and TEM images of substrate structural morphological changes showed that SPORL pretreatment resulted in fiber separation, where SPORL high pH (4.2) pretreatment exhibited better fiber separation than SPORL low pH (1.9) pretreatment. Dilute acid pretreatment produced very poor fiber separation, consisting mostly of fiber bundles. The removal of almost all hemicelluloses in the dilute acid pretreated substrate did not overcome recalcitrance to achieve a high cellulose conversion when lignin removal was limited. SPORL high pH pretreatment removed more lignin but less hemicellulose, while SPORL low pH pretreatment removed about the same amount of lignin and hemicelluloses in lodgepole pine substrates when compared with dilute acid pretreatment. Substrates pretreated with either SPORL process had a much higher cellulose conversion than those produced with dilute acid pretreatment. Lignin removal in addition to removal of hemicellulose in SPORL pretreatment plays an important role in improving the cellulose hydrolysis of the substrate.

Evaluation of pretreatment methods in improving the enzymatic saccharification of cellulosic materials

The effectiveness of alkali, acid and chlorite pretreatment of lignocellulosic feedstocks for improving the enzymatic saccharification of cellulose has been evaluated. The feedstocks such as Corncob, Prosopis juliflora and Lantana camara were pretreated with varied concentration of sulfuric acid, sodium hydroxide and sodium chlorite at 121 °C for 15–60 min. Among different methods used, chlorite pretreatment removed maximum lignin with ∼90% (w/w) residual holocellulose content in all the substrates tested. Moreover, irrespective of the substrates used, the chlorite treated substrates were enzymatically saccharified from 86.4% to 92.5% (w/w). While, the alkali treated substrates containing 66.0–76.0% (w/w) holocellulose could be enzymatically saccharified up to 55% (w/w). The acid pretreated substrates were found to contain almost 54–62% (w/w) holocellulose, which on enzymatic hydrolysis could result in 39.5–48% (w/w) saccharification.

Full-scale On-farm Pretreatment of Perennial Grasses with Dilute Acid for Fuel Ethanol Production

Biorefineries that rely on lignocellulosic feedstocks require dependable and safe methods for storing biomass. Storing biomass wet in the presence of sulfuric acid and the absence of oxygen has been shown to preserve carbohydrates and enhance cellulose conversion but has not been demonstrated at farm-scale. To that end, switchgrass (Panicum virgatum L.) and reed canarygrass (Phalaris arundinacea L.) were pretreated with 18 N sulfuric acid with two methods: during bagging (on-line) and thoroughly mixed in a commercial feed mixer (mixed) and both stored for 90 days. The two methods, applied at rates from 28 to 54 g(kg DM)−1 not only helped to preserve biomass substrates under on-farm conditions (anaerobic, ambient temperature and pressure) through inhibition of microbial activity but also enhanced conversion of cellulose to ethanol by simultaneous saccharification and fermentation (SSF) using Saccharomyces cerevisiae. Acid-pretreated substrate yielded 19 and 7 percentage points higher ethanol conversion efficiencies than fresh reed canarygrass and switchgrass, respectively. The on-line method of pretreatment out-yielded the mixed method both as a preservative and as an agent for enhanced cell wall degradation. This result was thought to be an outcome of more uniform acid application as indicated by the on-line method’s more consistent pH profile and decreased fermentation products, as compared to the mixed method. Although significant levels of acetate and lactate were present in the biomass following storage, concentrations were not sufficient to inhibit S. cerevisiae in SSFs with a 10% solids loading.

Separate hydrolysis and fermentation (SHF) of Prosopis juliflora, a woody substrate, for the production of cellulosic ethanol by Saccharomyces cerevisiae and Pichia stipitis-NCIM 3498

Prosopis juliflora (Mesquite) is a raw material for long-term sustainable production of cellulosics ethanol. In this study, we used acid pretreatment, delignification and enzymatic hydrolysis to evaluate the pretreatment to produce more sugar, to be fermented to ethanol. Dilute H 2SO 4 (3.0%, v/v) treatment resulted in hydrolysis of hemicelluloses from lignocellulosic complex to pentose sugars along with other byproducts such as furfural, hydroxymethyl furfural (HMF), phenolics and acetic acid. The acid pretreated substrate was delignified to the extent of 93.2% by the combined action of sodium sulphite (5.0%, w/v) and sodium chlorite (3.0%, w/v). The remaining cellulosic residue was enzymatically hydrolyzed in 0.05 M citrate phosphate buffer (pH 5.0) using 3.0 U of filter paper cellulase (FPase) and 9.0 U of β-glucosidase per mL of citrate phosphate buffer. The maximum enzymatic saccharification of cellulosic material (82.8%) was achieved after 28 h incubation at 50 °C. The fermentation of both acid and enzymatic hydrolysates, containing 18.24 g/L and 37.47 g/L sugars, with Pichia stipitis and Saccharomyces cerevisiae produced 7.13 g/L and 18.52 g/L of ethanol with corresponding yield of 0.39 g/g and 0.49 g/g, respectively.

Simultaneous saccharification and cofermentation of peracetic acid-pretreated biomass.

Previous work in our laboratories has demonstrated the effectiveness of peracetic acid for improving enzymatic digestibility of lignocellulosic materials. The use of dilute alkali solutions as a pre-pretreatment prior to peracetic acid lignin oxidation increased carbohydrate hydrolysis yields in a synergistic as opposed to additive manner. Deacetylation of xylan is easily achieved using dilute alkali solutions under mild conditions. In this article, we evaluate the effectiveness of peracetic acid combined with an alkaline pre-pretreatment through simultaneous saccharification and cofermentation (SSCF) of pretreated hybrid poplar wood and sugar cane bagasse. Respective ethanol yields of 92.8 and 91.9% of theoretical are achieved using 6% NaOH/15% peracetic acid-pretreated substrates and recombinant Zymomonas mobilis CP4/pZB5. Reduction of acetyl groups of the lignocellulosic materials is demonstrated following alkaline pre-pretreatments. Such processing may be helpful in reducing peracetic acid requirements. The influence of deacetylation is more significant in combined pretreatments using lower peracetic acid loadings.