In recent years, lignocellulosic biomass has emerged as one of the most versatile energy sources among the research community for the production of biofuels and value-added chemicals. However, biomass pretreatment plays an important role in reducing the recalcitrant properties of lignocellulose, leading to superior quality of target products in bioenergy production. Among existing pretreatment techniques, liquid hot water (LHW) pretreatment has several outstanding advantages compared to others including minimum formation of monomeric sugars, significant removal of hemicellulose, and positive environmental impacts; however, several constraints of LHW pretreatment should be clarified. This contribution aims to provide a comprehensive analysis of reaction mechanism, reactor characteristics, influencing factors, techno-economic aspects, challenges, and prospects for LHW-based biomass pretreatment. Generally, LHW pretreatment could be widely employed in bioenergy processing from biomass, but circular economy-based advanced pretreatment techniques should be further studied in the future to achieve maximum efficiency, and minimum cost and drawbacks.

Current utilization of reed scraps (RS) from reed pulping mills is just burning and burying, remaining the unused RS a major industry waste. The aim of this study was to valorize RS by utilizing it as feedstock for xylo-oligosaccharide (XOS) production. The RS was subjected to dilute acid, alkaline, and liquid hot water (LHW) pretreatments to get the pretreated RS, which was then subjected to xylanase and subsequently cellulase enzymatic to produce XOS and glucose as well. The highest yield of XOS and glucose were 0.144 g and 0.304 g / g of RS, respectively, which were obtained from the LHW pretreatment under the optimized conditions of 170 °C, 1:8 solid loadings and 30 min duration. Overall, by upgrading RS waste into XOS and glucose as value-added products of reed pulp production, we thereby paved a new way in this research to valorize the waste from reed pulping mills.

BACKGROUND: Liquid hot water (LHW) pretreatment has been considered as one of the most industrially viable and environment-friendly methods for facilitating the transformation of lignocelluloses into biofuels through biological conversion. However, lignin fragments in pretreatment hydrolysates are preferential to condense with each other and then deposit back onto cellulose surface under severe conditions. Particularly, lignin tends to relocate or redistribute under high-temperature LHW pretreatment conditions. The lignin residues on the cellulose surface would result in significant nonproductive binding of cellulolytic enzymes, and therefore negatively affect the enzymatic conversion (EC) of glucan in pretreated substrates. Although additives such as bovine serum albumin (BSA) and Tween series have been used to reduce nonproductive binding of enzymes through blocking the lignin, the high cost or non-biocompatibility of these additives limits their potential in industrial applications.
RESULTS: Here, we firstly report that a soluble soy protein (SP) extracted from inexpensive defatted soy powder (DSP) showed excellent performance in promoting the EC of glucan in LHW-pretreated lignocellulosic substrates. The addition of the SP (80 mg/g glucan) could readily reduce the cellulase (Celluclast 1.5 L®) loading by 8 times from 96.7 to 12.1 mg protein/g glucan and achieve a glucan EC of 80% at a hydrolysis time of 72 h. With the same cellulase (Celluclast 1.5 L®) loading (24.2 mg protein/g glucan), the ECs of glucan in LHW-pretreated bamboo, eucalyptus, and Masson pine substrates increased from 57%, 54% and 45% (without SP) to 87%, 94% and 86% (with 80 mg SP/g glucan), respectively. Similar effects were also observed when Cellic CTec2, a newer-generation cellulase preparation, was used. Mechanistic studies indicated that the adsorption of soluble SP onto the surface of lignin residues could reduce the nonproductive binding of cellulolytic enzymes to lignin. The cost of the SP required for effective promotion would be equivalent to the cost of 2.9 mg cellulase (Celluclast 1.5 L®) protein (or 1.2 FPU/g glucan), if a proposed semi-simultaneous saccharification and fermentation (semi-SSF) model was used.
CONCLUSIONS: Near-complete saccharification of glucan in LHW-pretreated lignocellulosic substrates could be achieved with the addition of the inexpensive and biocompatible SP additive extracted from DSP. This simple but remarkably effective technique could readily contribute to improving the economics of the cellulosic biorefinery industry.

The liquid hot water (LHW) pretreatment could be strengthened by acetic and lactic acids produced from the process. The synergistic effect of the mixed acid catalyst of lactic acid and acetic acid was investigated for the purpose of maximization of the overall C5 sugars yield. Individual acids (acetic and lactic acid) and mixed acid were used to strengthen the LHW pretreatment at different conditions. The results showed that the suitable conditions of mixed acid synergistic catalysis was at 180 °C for 60 min and 3 wt% mixed acid where the ratio of 40% (i.e. 0.40 in mass fraction of lactic acid in mixed acid). Response surface methodology (RSM) was applied to further optimize this process. The highest yield of C5 sugars of 93.83% according to theoretical predicted model, was close to the experiment value of 92.53% at 177 °C for 67 min and with the ratio of mixed acid of 40%.

UNASSIGNED: Hydrothermal pretreatment using liquid hot water (LHW) is capable of substantially reducing the cell wall recalcitrance of lignocellulosic biomass. It enhances the saccharification of polysaccharides, particularly cellulose, into glucose with relatively low capital required. Due to the close association with biomass recalcitrance, the structural change of the components of lignocellulosic materials during the pretreatment is crucial to understand pretreatment chemistry and advance the bio-economy. Although the LHW pretreatment has been extensively applied and studied, the molecular structural alteration during pretreatment and its significance to reduced recalcitrance have not been well understood.
UNASSIGNED: We investigated the effects of LHW pretreatment with different severity factors (log R0) on the structural changes of fast-grown poplar (Populus trichocarpa). With the severity factor ranging from 3.6 to 4.2, LHW pretreatment resulted in a substantial xylan solubilization by 50-77% (w/w, dry matter). The molecular weights of the remained hemicellulose in pretreated solids also have been significantly reduced by 63-75% corresponding to LHW severity factor from 3.6 to 4.2. In addition, LHW had a considerable impact on the cellulose structure. The cellulose crystallinity increased 6-9%, whereas its degree of polymerization decreased 35-65% after pretreatment. We found that the pretreatment severity had an empirical linear correlation with the xylan solubilization (R2 = 0.98, r = + 0.99), hemicellulose molecular weight reduction (R2 = 0.97, r = - 0.96 and R2 = 0.93, r = - 0.98 for number-average and weight-average degree of polymerization, respectively), and cellulose crystallinity index increase (R2 = 0.98, r = + 0.99). The LHW pretreatment also resulted in small changes in lignin structure such as decrease of β-O-4\' ether linkages and removal of cinnamyl alcohol end group and acetyl group, while the S/G ratio of lignin in LHW pretreated poplar residue remained no significant change compared with the untreated poplar.
UNASSIGNED: This study revealed that the solubilization of xylan, the reduction of hemicellulose molecular weights and cellulose degree of polymerization, and the cleavage of alkyl-aryl ether bonds in lignin resulted from LHW pretreatment are critical factors associated with reduced cell wall recalcitrance. The chemical-structural changes of the three major components, cellulose, lignin, and hemicellulose, during LHW pretreatment provide useful and fundamental information of factors governing feedstock recalcitrance during hydrothermal pretreatment.

Ensilage is a traditional way of preserving fresh biomass. However, in order to apply ensilage to the ethanol biorefinery, two parameters need to be evaluated: quantity and quality changes of the biomass; and its effects on bioconversion process. To study these two aspects, switchgrass harvested on three different time points (Early, mid and late fall) were used as feedstock. The early fall harvested biomass was ensiled at 5 moisture levels ranging from 30% to 70%. Silage of 40% moisture and 3 other raw switchgrass were pretreated with liquid hot water, followed by enzymatic hydrolysis as well as simultaneous saccharification and fermentation. After 21 days storage pH values of all silages decreased below 4.0 and the dry matter losses were less than 2.0%, and structural sugars contents did not change dramatically. Liquid hot water caused more hemicellulose dissolution in the silage than in unensiled switchgrass. However, ensilage also increased the risk of releasing more sugar degradation products; After enzymatic hydrolysis, silage obtained higher total glucose, xylose and galactose yields than raw materials; After simultaneous saccharification and fermentation, ethanol concentration in silage was 12.1 g/L, higher than the unensiled switchgrass (10.3 g/L, 9.7 g/L and 10.6 g/L for early, mid and late fall respectively). Our results suggest that ensilage helps increase pretreatment efficiency and sugar yield, which increases final ethanol production.
青贮是一种传统的生物质原料保存方法，广泛应用于纤维素乙醇炼制领域尚需要考察其对原料品质和下游乙醇转化过程的影响。文中以秋季 (初、中和末) 收割的柳枝稷为原料，通过青贮、高温水热 (LHW) 预处理、纤维素酶水解和同步糖化与发酵 (SSF) 实验对上述问题予以回答。结果显示，秋季初收割的柳枝稷以不同湿度青贮后pH 均小于4.0，干重损失小于2%，各主要成分与青贮前相比无明显变化；LHW 预处理中青贮样品半纤维素水解率普遍高于未贮存样品，但青贮同样使原料获得了更高的发酵抑制物产生水平；青贮柳枝稷葡萄糖、木糖和半乳糖产量 (预处理+酶水解) 高于未贮存柳枝稷；经过168 h 的SSF，青贮样品乙醇浓度为12.1 g/L，未贮存的秋季初、秋季中和秋季末柳枝稷为底物的浓度分别为10.3 g/L、9.7 g/L 和10.6 g/L。综上，青贮有助于提高柳枝稷LHW 预处理效率、酶水解率和乙醇产量。.

A bio-refinery process of wheat straw pulping solid residue (waste wheat straw, WWS) was established by combining prewashing and liquid hot water pretreatment (LHWP). The results showed that employing a prewashing step prior to the LHWP remarkably improved enzymatic glucose yields from 39.7% to 76.6%. Moreover, after 96h simultaneous saccharification and fermentation (SSF), identical ethanol yields of 0.41g/g-cellulose were obtained despite varied solid loadings (5-30%). Beyond ethanol, enzymatic post-hydrolysis of the prehydrolyzate effectively increased xylobiose and xylotriose yields from 15mg/g-WWS and 14mg/g-WWS to 53mg/g-WWS and 20mg/g-WWS, respectively. For mass balance, about 10.9tons raw WWS will be consumed to produce 1ton ethanol, in addition to producing 614.8kg xylooligosaccharides (XOS) containing 334.3kg xylobiose and 124.8kg xylotriose. The results demonstrated that the integrated process for the WWS bio-refinery is promising, based on value-adding co-production in addition to robust ethanol yields.

A novel combined pretreatment of deacetylation and liquid hot water (LHW) was invented which has been proved to be effective in increasing enzymatic hydrolysis yield of biomass. In order to further understand the effect of this new pretreatment process on biomass, the variation on structural properties including cellulose crystallinity index (CrI), specific surface area (SSA) and degree of polymerization (DP) before/after pretreatment and how these properties affected the enzymatic hydrolysis of biomass were explored. The improvement of pretreatment severity (PS) could increase CrI, SSA and reduce DP. Whereas the enhancement of degree of deacetylation could decrease SSA and DP. An optimal formula (E12Y=0.347(100-CrI)(-0.375)×(SSA)(0.203)×(1700-DP)(0.281)) was achieved to express the correlation between structural properties and enzymatic hydrolysis after 12h. The enzymatic yield was more sensitive to CrI than to SSA and DP.

The effect of prewashing process prior to the liquid hot water (LHW) pretreatment of high free ash content waste wheat straw (WWS) was investigated. It was found that prewashing process decreased the ash content of WWS greatly, from 29.48% to 9.82%. This contributed to the lower pH value of prehydrolyzate and higher xylan removal in the following LHW pretreatment. More importantly, the prewashing process effectively increased the cellulose enzymatic hydrolysis efficiency of pretreated WWS, from 53.04% to 84.15%. The acid buffering capacity (ABC) and cation exchange capacity (CEC) of raw and prewashed WWS were examined. The majority of free ash removal from WWS by prewashing resulted in the decrease of the ABC of the WWS from 211.74 to 61.81mmol/pH-kg, and potentially enhancing the efficiency of the follow-up LHW pretreatment.

BACKGROUND: In the bioconversion of lignocellulosic substrates, the adsorption behavior of cellulase onto lignin has a negative effect on enzymatic hydrolysis of cellulose, decreasing glucose production during enzymatic hydrolysis, thus decreasing the yield of fermentation and the production of useful products. Understanding the interaction between lignin and cellulase is necessary to optimize the components of cellulase mixture, genetically engineer high-efficiency cellulase, and reduce cost of bioconversion. Most lignin is not removed during liquid hot water (LHW) pretreatment, and the characteristics of lignin in solid substrate are also changed. To understand the interactions between cellulase and lignin, this study investigated the change in the characteristics of lignin obtained from corn stover, as well as the behavior of cellulase adsorption onto lignin, under various severities of LHW pretreatment.
RESULTS: LHW pretreatment removed most hemicellulose and some lignin in corn stover, as well as improved enzymatic digestibility of corn stover. After LHW pretreatment, the molecular weight of lignin obviously increased, whereas its polydispersity decreased and became more negative. The hydrophobicity and functional groups in lignin also changed. Adsorption of cellulase from Penicillium oxalicum onto lignin isolated from corn stover was enhanced after LHW pretreatment, and increased under increasing pretreatment severity. Different adsorption behaviors were observed in different lignin samples and components of cellulase mixtures, even in different cellobiohydrolases (CBHs), endo-beta-1, 4-glucanases (EGs). The greatest reduction in enzyme activity caused by lignin was observed in CBH, followed by that in xylanase and then in EG and β-Glucosidase (BGL). The adsorption behavior exerted different effects on subsequent enzymatic hydrolysis of various biomass substrates. Hydrophobic and electrostatic interactions may be important factors affecting different adsorption behaviors between lignin and cellulase.
CONCLUSIONS: LHW pretreatment changed the characteristics of the remaining lignin in corn stover, thus affected the adsorption behavior of lignin toward cellulase. For different protein components in cellulase solution from P. oxalicum, electrostatic action was a main factor influencing the adsorption of EG and xylanase onto lignin in corn stover, while hydrophobicity affected the adsorption of CBH and BGL onto lignin.