Non-productive Adsorption Of Cellulase Research Articles

Unveiling the mechanisms of mixed surfactant synergy in passivating lignin-cellulase interactions during lignocellulosic saccharification

Surfactants can synergistically enhance the enzymatic hydrolysis of lignocellulosic biomass, achieving higher sugar yields at lower enzyme loading. However, the exact mechanism by which mixed surfactants passivate lignin-cellulase interactions is not fully understood. This study found that the combination of ternary non-ionic and cationic surfactants (Tween 60, Triton X-114, and CTAB) significantly reduced the non-productive adsorption of lignin, with decreases of 35.4%-55.4% in equilibrium adsorption (We, 23.2mg/g) compared to the single surfactant and the control. Meanwhile, mixed surfactants disrupted the entropy-enthalpy co-driven process for non-productive cellulase adsorption while promoting the desorption process. Non-ionic surfactants mainly contributed to reducing the hydrophobic interactions between lignin and cellulases. Positively charged CTAB enabled nonionic surfactants to form stronger H-bonds with lignin by electrophilic modification, and Triton X-114 increased van der Waals forces. Although surfactant-modified lignin exhibited lower hydrophobicity, zeta potential, and a more stable hydrogen bond network, the inhibitory effects of lignin-cellulase interactions by mixed surfactants were susceptible to lignin properties. According to the structure-activity relationship analysis (R2>0.80), the main influencing factors included particle size, aliphatic/phenolic OH group contents, contact angle, and zeta potential of lignin. The study on the synergistic passivation of lignin-cellulase interactions by multi-component surfactant systems provides some theoretical insights for selecting and customarily designing effective additives for efficient enzymatic hydrolysis in lignocellulosic biorefineries.

The role of glycosylation in non-productive adsorption of cellulase to lignin isolated from pretreated corn stover

Glycosylation, a general post-translational modification for fungal cellulase, has been shown to affect cellulase binding to its substrate. However, the exact impact of glycosylation on cellulase-lignin interaction remain unclear. Here, we demonstrated that the lignin isolated from tetrahydrofuran-pretreated corn stover exhibits strong adsorption capability to cellulase due to its negatively charged and porous structure. For the cellulases with varying glycosylation levels, the less-glycosylated protein showed high adsorption capability to lignin, and that trend was observed for the main cellulase components secreted by Penicillium oxilicum, including endoglucanase PoCel5B, cellobiohydrolase PoCel7A-2, and β-glucosidase PoBgl1. Additionally, N-glycan sites and motifs were examined using mass spectrometry, and protein structures with N-glycans were constructed, where PoBgl1 and PoCel7A-2 contained 13 and 1 glycosylated sites respectively. The results of molecular dynamics simulations indicated that the N-glycans impacted on the solvent-accessible surface area and secondary structure of protein, and the binding conformation of lignin fragment on cellulase, resulting in a decrease in binding energy (14kcal/mol for PoBgl1 and 13kcal/mol for PoCel7A-2), particularly for van der Waals and electrostatic interaction. Those findings suggested that glycosylation negatively impacted the lignin-cellulase interaction, providing a theoretical basis for the rational engineering of enzymes to reduce lignin-enzyme interaction.

The Limitation of Unproductive Binding of Cellulases to Lignin by Ozone Pretreatment

The limitation of enzymatic saccharification of lignocellulose is attributed to the nonproductive adsorption between lignin and cellulase. This study aims to investigate the effects of ozone pretreatment on the physical structure and chemical properties of milled wood lignin (MWL). The objective is to reduce the non-productive adsorption of cellulase on lignin. The structure–activity relationship between the physical structure of MWL and the occurrence of nonproductive adsorption was analysed using two-dimensional heteronuclear single quantum coherence–nuclear magnetic resonance (HSQC-NMR) and phosphorus-31 nuclear magnetic resonance spectrum (31P-NMR), etc. The results indicate that ozone pretreatment resulted in a decrease in the phenolic hydroxyl content and S/G ratio, an increase in the carboxyl content, and a negative zeta potential of MWL. The maximum adsorption capacity decreased from 25.77 mg/g to 10.09 mg/g, the Langmuir constant decreased from 13.86 mL/mg to 10.11 mL/mg, and the binding strength decreased from 357.14 mL/g to 102.04 mL/g, as determined by Langmuir isothermal adsorption. This suggests that ozone pretreatment resulted in a reduction in the hydrophobicity of lignin and a weakening of the electrostatic attraction between lignin and cellulase, thereby effectively reducing the non-productive adsorption of cellulase on lignin. This study provides an environmentally friendly pretreatment technique and comprehensively analyses the structural changes of ozone-treated MWL. These findings contribute to a deeper understanding of the interaction between lignin and cellulase.

Aqueous amine enables sustainable monosaccharide, monophenol, and pyridine base coproduction in lignocellulosic biorefineries

Thought-out utilization of entire lignocellulose is of great importance to achieving sustainable and cost-effective biorefineries. However, there is a trade-off between efficient carbohydrate utilization and lignin-to-chemical conversion yield. Here, we fractionate corn stover into a carbohydrate fraction with high enzymatic digestibility and reactive lignin with satisfactory catalytic depolymerization activity using a mild high-solid process with aqueous diethylamine (DEA). During the fractionation, in situ amination of lignin achieves extensive delignification, effective lignin stabilization, and dramatically reduced nonproductive adsorption of cellulase on the substrate. Furthermore, by designing a tandem fractionation-hydrogenolysis strategy, the dissolved lignin is depolymerized and aminated simultaneously to co-produce monophenolics and pyridine bases. The process represents the viable scheme of transforming real lignin into pyridine bases in high yield, resulting from the reactions between cleaved lignin side chains and amines. This work opens a promising approach to the efficient valorization of lignocellulose.

A Novel Kinetic Modeling of Enzymatic Hydrolysis of Sugarcane Bagasse Pretreated by Hydrothermal and Organosolv Processes.

Lignocellulosic biomasses have a complex and compact structure, requiring physical and/or chemical pretreatments to produce glucose before hydrolysis. Mathematical modeling of enzymatic hydrolysis highlights the interactions between cellulases and cellulose, evaluating the factors contributing to reactor scale-up and conversion rates. Furthermore, this study evaluated the influence of two pretreatments (hydrothermal and organosolv) on the kinetics of enzymatic hydrolysis of sugarcane bagasse. The kinetic parameters of the model were estimated using the Pikaia genetic algorithm with data from the experimental profiles of cellulose, cellobiose, glucose, and xylose. The model considered the phenomenon of non-productive adsorption of cellulase on lignin and inhibition of cellulase by xylose. Moreover, it included the behavior of cellulase adsorption on the substrate throughout hydrolysis and kinetic equations for obtaining xylose from xylanase-catalyzed hydrolysis of xylan. The model for both pretreatments was experimentally validated with bagasse concentration at 10% w/v. The Plackett-Burman design identified 17 kinetic parameters as significant in the behavior of process variables. In this way, the modeling and parameter estimation methodology obtained a good fit from the experimental data and a more comprehensive model.

Employing Cationic Kraft Lignin as Additive to Enhance Enzymatic Hydrolysis of Corn Stalk.

A water-soluble cationic kraft lignin (named JLQKL50), synthesized by combining quaternization and crosslinking reactions, was used as an additive to enhance the enzymatic hydrolysis of dilute-alkali-pretreated corn stalk. The chemical constitution of JLQKL50 was investigated by Fourier transform infrared spectroscopy, 1H nuclear magnetic resonance (NMR) and 13C NMR spectroscopy, and elemental analysis. The enzymatic hydrolysis efficiency of corn stalk at solid content of 10% (w/v) was significantly improved from 70.67% to 78.88% after 24 h when JLQKL50 was added at a concentration of 2 g/L. Meanwhile, the enzymatic hydrolysis efficiency after 72 h reached 91.11% with 10 FPU/g of cellulase and 97.92% with 15 FPU/g of cellulase. In addition, JLQKL50 was found capable of extending the pH and temperature ranges of enzymatic hydrolysis to maintain high efficiency (higher than 70%). The decrease in cellulase activity under vigorous stirring with the addition of JLQKL50 was 17.4%, which was much lower than that (29.7%) without JLQKL50. The addition of JLQKL50 reduced the nonproductive adsorption of cellulase on the lignin substrate and improved the longevity, dispersity, and stability of the cellulase by enabling electrostatic repulsion. Therefore, the enzymatic hydrolysis of the corn stalk was enhanced. This study paves the way for the design of sustainable lignin-based additives to boost the enzymatic hydrolysis of lignocellulosic biomass.

Exploring how lignin structure influences the interaction between carbohydrate-binding module and lignin using AFM

Nonproductive adsorption of cellulase onto the residual lignin in substrate seriously hinders the enzymatic hydrolysis. To understand how lignin structure affects lignin-cellulase interaction, the carbohydrate-binding module (CBM) functionalized atomic force microscope tip was used to measure CBM-lignin interaction by single-molecule dynamic force spectroscopy in this work. The results showed that sulfonated lignin (SL) has the greatest adhesion force to CBM (4.74 nN), while those of masson pine milled wood lignin (MWL), poplar MWL and herbaceous MWLs were 2.85, 1.03 and 0.27-0.61 nN, respectively. It provides direct quantitative evidence for the significance of lignin structure on lignin-cellulase interaction. The CBM-MWLs interaction decreased sharply to 0.054-0.083 nN while SL was added, indicating the primary mechanism of SL promoting lignocellulose hydrolysis was significantly reducing the nonproductive adsorption of substrate lignin on cellulase. Finally, the "competitive adsorption" mechanism was proposed to interpret why SL effectively promotes the enzymatic hydrolysis of lignin-containing substrates.

Alleviating Nonproductive Adsorption of Lignin on CBM through the Addition of Cationic Additives for Lignocellulosic Hydrolysis.

The nonproductive adsorption of cellulase onto lignin significantly inhibited the enzymatic hydrolysis of lignocellulosic biomass. In this study, we constructed a rapid fluorescence detection (RFD) system, and using this system, we demonstrated that the addition of cationic additives DTAB or polyDADMAC greatly increased the partition coefficients of cellulose/lignin, reduced nonproductive adsorption, and enhanced the hydrolysis efficiency of lignocellulose compared to those of Tweens or PEGs. Moreover, the addition of polyDADMAC and DTAB increased the glucose yield released from the mixture of Avicel and AICS-lignin (MCL) by 16.9 and 20.6%, respectively, and reduced the inhibition rate of lignin by 16.9 and 20.7%, respectively. Interestingly, polyDADMAC or DTAB treatment performed more effectively for the enzymatic hydrolysis of pretreated lignocellulosic biomass, compared with MCL. We confirmed that the reduced hydrophobicity and increased zeta potential of lignin cocontribute to the dampening nonproductive adsorption of lignin. In particular, the zeta potential values of lignin and the partition coefficients of Avicel/lignin with the addition of additives showed a good correlation, suggesting that electrostatic force also plays a crucial role in the adsorbing of cellulase on lignin. This work will be conducive to decreasing the nonproductive binding of cellulase onto lignin and enhancing cellulose conversion.

Polystyrene sulfonate is effective for enhancing biomass enzymatic saccharification under green liquor pretreatment in bioenergy poplar

BackgroundWater-soluble lignin (particularly lignosulfonate, LS) has been well documented for its significance on enzymatic saccharification of lignocellulose, though the promotion mechanism has not been fully understood. Much attention has been paid to natural lignin or its derivatives. The disadvantage of using natural lignin-based polymers as promoting agents lies in the difficulty in tailor-incorporating functional groups due to their complex 3D structures. To further improve our understanding on the promotion mechanism of water-soluble lignin in the bioconversion of lignocellulose and to pursue better alternatives with different skeleton structures other than natural lignin, herein we reported a synthetic soluble linear aromatic polymer, sodium polystyrene sulfonate (PSS), to mimic LS for enhancing the efficiency of enzymatic saccharification.ResultsThe role of PSS in enzymatic saccharification of pure cellulose and green liquor-pretreated poplar (GL-P) was explored by analyzing substrate enzymatic digestibility (SED) under different addition dosages and various pH media, along with LS for comparison. At the cellulase loading of 13.3 FPU/g-glucan, the glucose yield of GL-P increased from 53% for the control to 81.5% with PSS addition of 0.1 g/g-substrate. It outperformed LS with the addition of 0.2 g/g-substrate by 6.3%. In the pH range from 4.5 to 6, PSS showed a positive effect on lignocellulose saccharification with the optimum pH at 4.8, where the most pronounced SED of GL-P was achieved. The underlying mechanism was unveiled by measuring zeta potential and using Quartz Crystal Microbalance (QCM) and Multi-parametric Surface Plasmon Resonance (MP-SPR). The results confirmed that the complexes of cellulase and PSS were conjugated and the negatively supercharged complexes reduced non-productive binding effectively along with the improved saccharification efficiency. The thickness of PSS required to block the binding sites of cellulase film was less than half of that of LS, and the PSS adlayer on cellulase film is also more hydrated and with a much lower shear modulus than LS adlayer.ConclusionsPSS as LS analogue is effective for enhancing the biomass enzymatic saccharification of GL-pretreated poplar. PSS exhibited a severer inhibition on the enzymatic saccharification of pure cellulose, while a more positive effect on bioconversion of lignocellulose (GL-P) than LS. In addition, a much lower dosage is required by PSS. The dynamic enzymatic hydrolysis indicated PSS could prolong the processive activity of cellulase. The valid data stemmed from QCM and SPR expressed that PSS bound to cellulases and the as-formed complexes reduced the non-productive adsorption of cellulase onto substrate lignin more efficiently than LS due to its flexible skeleton and highly hydrated structure. Therefore, PSS is a promising alternative promoting agent for lignocellulose saccharification. From another perspective, the synthetic lignin mimics with controllable structures enable us to reach an in-depth understanding of the promotion mechanism of soluble lignins on enzymatic saccharification.

Comparison of sulfomethylated lignin from poplar and masson pine on cellulase adsorption and the enzymatic hydrolysis of wheat straw

In this work, effects of sulfomethylated lignins (SLs) prepared from masson pine (SLM) and poplar (SLP) on enzymatic hydrolysis and cellulase-lignin interaction were comparatively investigated. The results showed that both SLM and SLP significantly promoted the substrate enzymatic digestibility. The total sugar yield increased from 38.6% to 74.4% and∼100%, respectively at 10 FPU/g-cellulose of cellulase dosage. The protein content in hydrolysate linearly increased with the addition of SL (0 - 1.6g/g-substrate lignin), which suggested the competitive adsorption of cellulase may occur to substrate lignin and SLs. Further structural analysis of lignin revealed the high S/(V+H) ratio was directly related to the high enzymatic saccharification efficiency. The strong interaction between SL and cellulase decreased the nonproductive adsorption of cellulase onto substrate lignin and increased the accessibility of cellulase to carbohydrate, which was considered to be the key factor for the improvement of substrate enzymatic digestibility.

Cellulose induced protein 1 (Cip1) from Trichoderma reesei enhances the enzymatic hydrolysis of pretreated lignocellulose

BackgroundTrichoderma reesei is currently the main strain for the commercial production of cellulase. Cellulose induced protein 1 (Cip1) is one of the most abundant proteins in extracellular proteins of T. reesei. Reported literatures about Cip1 mainly focused on the regulation of Cip1 and its possible enzyme activities, but the effect of Cip1 on the enzymatic hydrolysis of lignocellulose and possible mechanism have not still been reported.ResultsIn this study, Cip1 from T. reesei was cloned, expressed and purified, and its effects on enzymatic hydrolysis of several different pretreated lignocellulose were investigated. It was found that Cip1 could promote the enzymatic hydrolysis of pretreated lignocellulose, and the promoting effect was significantly better than that of bovine serum albumin (BSA). And especially for the lignocellulosic substrate with high lignin content such as liquid hot water pretreated corn stover and corncob residue, the promoting effect of Cip1 was even better than that of the commercial cellulase when adding equal amount protein. It was also showed that the metal ions Zn2+ and Cu2+ influenced the promoting effect on enzymatic hydrolysis. The Cip1 protein had no lyase activity, but it could destroy the crystal structure of cellulose and reduce the non-productive adsorption of cellulase on lignin, which partly interpreted the promoting effect of Cip1 on enzymatic hydrolysis of lignocellulose.ConclusionThe Cip1 from T. reesei could significantly promote the enzymatic hydrolysis of pretreated lignocellulose, and the promotion of Cip1 was even higher than that of commercial cellulase in the enzymatic hydrolysis of the substrates with high lignin content. This study will help us to better optimize cellulase to improve its ability to degrade lignocellulose, thereby reducing the cost of enzymes required for enzymatic hydrolysis.

Understanding the promoting effect of non-catalytic protein on enzymatic hydrolysis efficiency of lignocelluloses

Lignin deposits formed on the surface of pretreated lignocellulosic substrates during acidic pretreatments can non-productively adsorb costly enzymes and thereby influence the enzymatic hydrolysis efficiency of cellulose. In this article, peanut protein (PP), a biocompatible non-catalytic protein, was separated from defatted peanut flour (DPF) as a lignin blocking additive to overcome this adverse effect. With the addition of 2.5 g/L PP in enzymatic hydrolysis medium, the glucose yield of the bamboo substrate pretreated by phenylsulfonic acid (PSA) significantly increased from 38 to 94% at a low cellulase loading of 5 FPU/g glucan while achieving a similar glucose yield required a cellulase loading of 17.5 FPU/g glucan without PP addition. Similar promotion effects were also observed on the n-pentanol-pretreated bamboo and PSA-pretreated eucalyptus substrates. The promoting effect of PP on enzymatic hydrolysis was ascribed to blocking lignin deposits via hydrophobic and/or hydrogen-bonding interactions, which significantly reduced the non-productive adsorption of cellulase onto PSA lignin. Meanwhile, PP extraction also facilitated the utilization of residual DPF as the adhesive for producing plywood as compared to that without protein pre-extraction. This scheme provides a sustainable and viable way to improve the value of woody and agriculture biomass.Peanut protein, a biocompatible non-catalytic protein, can block lignin, improve enzymatic hydrolysis efficiency and thereby facilitate the economics of biorefinery.Graphical abstract

Features correlated to improved enzymatic digestibility of corn stover subjected to alkaline hydrogen peroxide pretreatment

As one of the leading pretreatment approaches, alkaline hydrogen peroxide (AHP) pretreatment can enhance the enzymatic digestibility of lignocellulose significantly. In this study, the glucan conversion of AHP pretreated corn stover (CS) without and with water-wash were 28.4% and 50.0% higher than that of raw material, respectively. In order to systematically understand its mechanism, analyses of the features of AHP pretreated and raw CS, such as specific surface area, crystallinity, zeta potential, water holding capacity and swelling capacity and others were performed. The weight-average molecular weight (Mw) of the sugars in the hydrolysate and the particle size distribution of the hydrolysis residue were also analyzed. These results explained why AHP-CS was more conducive to enzymatic hydrolysis. The deeper reason was that the removal of lignin and the destruction of hydrogen bonds within cellulose and hemicellulose increased the accessibility of cellulose and reduced the non-productive adsorption of cellulase, which significantly improved the enzymatic digestibility.

Effects of Biosurfactants on Enzymatic Saccharification and Fermentation of Pretreated Softwood.

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, Pseudomonas aeruginosa 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 Saccharomyces cerevisiae 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.

Effects of Rhamnolipids on Enzymatic Hydrolysis of Bamboo Biomass and Mechanism

The lignin present in lignocellulose seriously affects the efficiency of cellulose enzymatic hydrolysis. In addition, lignin adsorbs high-cost cellulase, causing greater economic losses. Lignin can also disturb the site of action of cellulase and reduce the efficiency of hydrolysis. Therefore, if lignin is removed or surface modified before cellulose enzymatic hydrolysis, the enzymatic hydrolysis efficiency of lignocellulosic biomass will be greatly improved. In this paper, the cellulose enzymatic properties of bamboo biomass being treated with dilute acid and alkaline under the intervention of biosurfactant rhamnolipid were evaluated. The effects of rhamnolipids on the adsorption characterization of cellulose on pretreated bamboo were studied. Besides, the inter-communication between rhamnolipids and cellulose was investigated by fluorescence probe. The results showed that rhamnolipids could have a positive effect on the enzymatic hydrolysis of bamboo biomass by reducing the non-productive adsorption of cellulase on the surface of lignocellulose. The outcome illustrated that cellulase could be combined with rhamnolipids micelles, participating in the formation of rhamnolipids micelles, thereby increasing the internal hydrophobicity of the micelles, but could not change the properties of rhamnolipids micelles higher than one CMC (Critical Micelle Concentration). It can be seen that the interaction between rhamnolipids and cellulase is beneficial to enhance the stability and enzymatic activity of cellulase, thereby improving the enzymatic hydrolysis efficiency of cellulose in biomass. Based on these results, a theoretical knowledge about the mechanism of enhancing the enzymatic hydrolysis efficiency of lignocellulose by biosurfactants rhamnolipids is provided.

Comparison of nonproductive adsorption of cellulase onto lignin isolated from pretreated lignocellulose

Lignin, as a main factor inhibiting the enzymatic hydrolysis of lignocellulose, was investigated by comparing lignin isolated from untreated and pretreated lignocellulose to elucidate the differences in the enzyme adsorption ability. The nonproductive adsorption of cellulase on lignin was investigated by using a quartz crystal microbalance with dissipation. The results indicated that more cellulase adsorbed on the lignin isolated from autohydrolysis and green liquor-pretreated lignocellulose than on protolignin. Higher temperature and enzyme concentration increased the initial adsorption rate and the amount of enzyme adsorbed on lignin. A dynamic equation for enzyme adsorption on lignin was established. The adsorption of enzymes on lignin increased as the content of phenolic hydroxyl groups, and β-β and β-5 linkages in the lignin increased, attributed to enhanced hydrogen bonding and hydrophobic interaction. Consequently, inhibition of the enzymatic hydrolysis of cellulose was promoted by a higher phenolic hydroxyl group, β-β, and β-5 content. The adsorption of the enzyme components on lignin followed the order: β-glucosidase > xylanase > endo-glucanase > cellobiohydrolase, as revealed by polyacrylamide gel electrophoresis analysis. Electrostatic interaction increased the adsorption of β-glucosidase and xylanase on lignin, while synergistic hydrophobic interaction and hydrogen bonding increased the adsorption of endo-glucanase and cellobiohydrolase on lignin. The results indicate that the adsorption of enzymes on lignin is affected by the structure of lignin and the composition of the enzymes. The findings of the current study provide fundamental knowledge for improving the enzymatic conversion of lignocellulose.

Effect of lignin isolated from p-toluenesulfonic acid pretreatment liquid of sugarcane bagasse on enzymatic hydrolysis of cellulose and cellulase adsorption

Abstract During lignocellulosic enzymatic hydrolysis, lignin is generally considered negative. To investigate the relation between lignin profiles and the effect of lignin on hydrolysis, in this study, the enzymatic hydrolysis efficiency and cellulase adsorption amount of Avicel and PB (p-toluenesulfonic acid (p-TsOH) treated sugarcane bagasse) were firstly determined before and after adding the p-TsOH lignin (isolated lignin from PB). Subsequently, the particle size, crystallinity, hydrogen bonds, electrostatic charges and hydrophobicity of PB and Avicel, and the chemical bonds of p-TsOH lignin were measured. The results indicated that adding the p-TsOH lignin inhibited Avicel’s hydrolysis, but increased the cellulase adsorption. This might be because after adding p-TsOH lignin, the lignin adsorbed more cellulase and the cellulase molecules on Avicel decreased. For PB, adding the p-TsOH lignin reduced the cellulase adsorption, but did not impact the hydrolysis efficiency. This might be due to the decrease in non-productive adsorption of cellulase.

Enhancement of enzyme hydrolysis by increasing the zeta potential to reduce non-productive cellulase adsorption on sugarcane bagasse treated with liquid hot water

The enhancement of enzymatic hydrolysis is important for the biorefinery industry of lignocellulose. Changing the pH of hydrolysis is a simple and direct way to improve hydrolysis efficiencies. In this study, the enzymatic hydrolysis efficiencies of sugarcane bagasse (SCB) treated with liquid hot water (LHW) were 56.7% and 65.5% at pHs of 4.8 and 5.5, respectively. The result of cellulase adsorption on the LHW treated SCB showed that the non-productive adsorption was smaller at pH 5.5, which might tend to enhance hydrolysis. The surface hydrophobicity of lignin was larger at pH 5.5. This suggested that the hydrophobic interaction was not dominant because a strong hydrophobicity force can cause more non-productive adsorption of cellulase with lignin. At pH 5.5, the surface negative charges of lignin and cellulase increased. Therefore, the electrostatic repulsive force between lignin and cellulase increased, leading to less of the non-productive adsorption of cellulase on lignin. In addition, the cellulase desorption from the LHW treated SCB also increased at pH 5.5. This was beneficial in increasing the possibility of cellulase re-adsorption in new binding sites on cellulose and promoting enzyme hydrolysis efficiency.

Exploring why sodium lignosulfonate influenced enzymatic hydrolysis efficiency of cellulose from the perspective of substrate\u2013enzyme adsorption

BackgroundCellulase adsorbed on cellulose is productive and helpful to produce reducing sugars in enzymatic hydrolysis of lignocellulose; however, cellulase adsorbed on lignin is non-productive. Increasing productive adsorption of cellulase on cellulose would be beneficial in improving enzymatic hydrolysis. Adding lignin that was more hydrophilic in hydrolysis system could increase productive adsorption and promote hydrolysis. However, the effect mechanism is still worth exploring further. In this study, lignosulfonate (LS), a type of hydrophilic lignin, was used to study its effect on cellulosic hydrolysis.ResultsThe effect of LS on the enzymatic hydrolysis of pure cellulose (Avicel) and lignocellulose [dilute acid (DA) treated sugarcane bagasse (SCB)] was investigated by analyzing enzymatic hydrolysis efficiency, productive and non-productive cellulase adsorptions, zeta potential and particle size distribution of substrates. The result showed that after adding LS, the productive cellulase adsorption on Avicel reduced. Adding LS to Avicel suspension could form the Avicel–LS complexes. The particles were charged more negatively and the average particle size was smaller than Avicel before adding LS. In addition, adding LS to cellulase solution formed the LS–cellulase complexes. For DA-SCB, adding LS decreased the non-productive cellulase adsorption on DA-SCB from 3.92 to 2.99 mg/g lignin and increased the productive adsorption of cellulase on DA-SCB from 2.00 to 3.44 mg/g cellulose. Besides, the addition of LS promoted the formation of LS–lignin complexes and LS–cellulase complexes, and the complexes had more negative charges and smaller average sizes than DA-SCB lignin and cellulase particles before adding LS.ConclusionsIn this study, LS inhibited Avicel’s hydrolysis, but enhanced DA-SCB’s hydrolysis. This stemmed from the fact that LS could bind cellulase and Avicel, and occupied the binding sites of cellulase and Avicel. Thus, a decreased productive adsorption of cellulase on Avicel arose. Regarding DA-SCB, adding LS, which enhanced hydrolysis efficiency of DA-SCB, increased the electrostatic repulsion between DA-SCB lignin and cellulase, and therefore, decreased non-productive adsorption of cellulase on DA-SCB lignin and enhanced productive adsorption of cellulase on DA-SCB cellulose.Graphical abstract

Comparative study on the properties of lignin isolated from different pretreated sugarcane bagasse and its inhibitory effects on enzymatic hydrolysis

Five sugarcane bagasse lignin samples, namely, dilute sulfuric acid (DSAL), sodium hydroxide (SHL), ethanol (EL), hot liquid water (HLWL)-pretreated residual solids, and raw material (cellulolytic enzyme lignin, CEL), were extracted. Comparative studies on the physicochemical properties of isolated lignin, nonproductive adsorption of cellulase by lignin, and its effect on enzymatic hydrolysis was performed. Results showed that the molecular weight and homogeneity of lignin remarkably decreased after pretreatment compared with CEL. Lignin with low negative zeta potential, high phenolic hydroxyl group content and hydrophobicity exhibited strong nonproductive adsorption performance to cellulase. This phenomenon was positively correlated with it's inhibitory effect on enzymatic hydrolysis. Compared with the control (without lignin), the Avicel conversion rate (40mg lignin/200mg Avicel) decreased by 10.74%, 9.28%, 8.73%, 4.22%, and 2.80% after digestion of Avicel for 72h with the presence of EL, SHL, CEL, HLWL, and DSAL, respectively.