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Due to the poor enzyme thermal stability, the efficient conversion of high crystallinity cellulose into glucose in aqueous phase over 50 °C is challenging. Herein, an enzyme‐induced MOFs encapsulation of β‐glucosidase (β‐G) strategy was proposed for the first time. By using various methods, including SEM, XRD, XPS, NMR, FTIR and BET, the successful preparation of a porous channel‐type flower‐like enzyme complex (β‐G@MOFs) was confirmed. The prepared enzyme complex (β‐G@MOFs) materials showed improved thermal stability (from 50 °C to 100 °C in the aqueous phase) and excellent resistance to ionic liquids (the reaction temperature was as high as 110 °C) compared to the free enzyme (β‐G). Not only the catalytic hydrolysis of cellulose by single enzyme (β‐G) in ionic liquid was realized, but also the high‐temperature continuous reaction performance of the enzyme was significantly improved. Benefiting from the significantly improved heat resistance, the β‐G@MOFs exhibited 32.1 times and 34.2 times higher enzymatic hydrolysis rate compared to β‐G for cellobiose and cellulose substrates, respectively. Besides, the catalytic activity of β‐G@MOFs was retained up to 86 % after five cycles at 110 °C. This was remarkable because the fixation of the enzyme by the MOFs ensured that the folded structure of the enzyme would not expand at high temperatures, allowing the native conformation of the encapsulated protein well‐maintained. Furthermore, we believe that this structural stability was caused by the confinement of flower‐like porous MOFs. [ABSTRACT FROM AUTHOR]

Some zwitterionic surfactants exhibit upper critical solution temperature (UCST) in aqueous solutions. For the zwitterionic surfactant solution mixed with cellulase, when its temperature is below UCST, the cellulase can be recovered by coprecipitation with zwitterionic surfactant. In this work, 3-(Hexadecyldimethylammonio) propanesulfonate (SB3-16) was selected to enhance the enzymatic hydrolysis of lignocelluloses and recover the cellulase. After adding 2 mmol/L of SB3-16, the enzymatic digestibility of eucalyptus pretreated by dilute acid (Eu-DA) and by sulfite (Eu-SPORL) increased from 27.9% and 35.1% to 72.6% and 89.7%, respectively. The results showed that SB3-16 could reduce the non-productive adsorption of cellulase on hydrophobic interface, while it did not significantly inhibit the activity of cellulase. For the solution contained 1 wt% SB3-16 and 200 mg protein/L CTec2 cellulase, 55.2% of protein could be recovered by cooling. The filter paper activity of the recovered cellulase was 1.93 FPU/mg protein, which was 95.8% of its initial activity. [ABSTRACT FROM AUTHOR]

Effect of cationic surfactants alkyltrimethylammonium bromide (CTAB) with varied alkyl chain lengths on the enzymatic hydrolysis of Avicel and the surface charge of cellulase was investigated. Enzymatic hydrolysis of Avicel increased linearly from 42.1 to 61.4 % with the increase of the concentration of cetyltrimethylammonium bromide (CTAB) logarithmically from 0.0001 to 0.01 mM, and reached a maximum value at the concentration of 0.01-0.03 mM. When the concentration was increased further, the cellulase solution became positively charged and the enzymatic hydrolysis of Avicel decreased rapidly. With the increasing alkyl chain length, CTAB provided more proton and neutralized the negative charge of cellulase more obviously. Therefore, the required concentration of CTAB could be less to enhance the enzymatic hydrolysis of Avicel. In addition, CTAB could enhance enzymatic hydrolysis efficiency of corncob at high solid content from 35.0 to 56.3 %; CTAB could reduce about 60 % of the cellulase loading in the enzymatic hydrolysis of corncob to obtain the same glucose yield. Effect of CTAB on the enzymatic hydrolysis of typical pretreated softwood and hardwood was also investigated. This study laid the foundation for using CTAB to recover cellulase, and provided the design direction for cellulase with higher activity and better stability by adjusting its hydrophilicity and chargeability. [ABSTRACT FROM AUTHOR]

[Display omitted] Cellulase was recycled by using pH-UCST-type LSB, and the process is sustainable while realizing the value-added utilization of industrial lignin. • LSB showed a pH-UCST-sensitive response and was recyclable. • Cellulase was efficiently recycled at higher pH by using LSB. • The value-added utilization of enzymatic hydrolysis lignin was realized. Recycling cellulase can reduce the cost of lignocellulosic enzymatic hydrolysis. Here, a lignin-grafted sulfobetaine (LSB) was first synthesized by grafting sulfobetaine (SB) on enzymatic hydrolysis lignin (EHL). LSB had a sensitive response of pH and temperature. LSB dissolved under the conditions of lignocellulosic enzymatic hydrolysis (pH 5.0, 50 °C). After hydrolysis, LSB co-precipitated with cellulase when lowering pH of the hydrolysate to 4.0 and cooling to 25 °C. When 3.0 g/L LSB-100 was added to the hydrolysis system of corncob residue (CCR), 70 % of amount of cellulase was saved. LSB had a remarkable response and stronger cellulase recovery capacity. This was attributed that carboxylate radical in LSB was protonated, and positive and negative ions of SB associated to form salt at 25 °C. This work provides a new idea for reducing the cost for preparing fermentable sugars from lignocellulose, and increasing the added value of EHL. [ABSTRACT FROM AUTHOR]

• Lignin-based surfactants LC and LCQ with various isoelectric points were synthesized. • The effect and mechanism of the pI of LCQ on the enzymatic hydrolysis were studied. • The SED of Eu-SPORL increased from 35.2% to 95.3% by adding LCQ-40. • LCQ could be efficiently recovered for the excellent pH-response performance. The isoelectric point (pI) of lignin-based surfactant is an important factor in the enhancement on the enzymatic hydrolysis of lignocellulose. In this work, lignin carboxylate (LC) and quaternary ammonium lignin carboxylates (LCQ-x, x%: the mass ratio of quaternizing agent to enzymatic hydrolysis lignin) with different isoelectric points were synthesized. LC or LCQ-x with pI significantly lower or higher than 4.8 reduced the non-productive adsorption of cellulase on lignin, but for the significant inhibitory effect on cellulase activity, their enhancements on the enzymatic hydrolysis of lignocellulose were not remarkable. However, LCQ-x with pI around 4.8 preserved the cellulase activity, and significantly reduced the non-productive adsorption of cellulase, therefore remarkably enhanced the enzymatic hydrolysis. 2 g/L LC, LCQ-40 (pI = 5.0) and LCQ-100 (pI = 9.2) increased the enzymatic digestibility of pretreated eucalyptus from 35.2% to 53.4%, 95.3% and 60.4% respectively. In addition, for the excellent pH-response performance, LCQ could be efficiently recovered after enzymatic saccharification. [ABSTRACT FROM AUTHOR]

The marriage of AFM, QCM and computational simulation quantitatively offers in-depth understanding of cellulase interaction with lignin, toward achieving an economic production of biofuels. We found that the long-range hydrophobic interaction firstly captures protein from solution and decreases the distance between cellulase and lignin so that other short-range interactions play a role in stabilizing the non-productive adsorption. [Display omitted] • Lignin-enzyme interactions were unmasked using a combination of atomic force microscopy and computational simulations. • The temperature sensitivity of cellulase adsorption behaviors is largely due to the changes in hydrophobic interactions. • The contribution of different long-range and short-range forces was revealed. • Hydrophobic interactions, hydrogen bonding, and steric effects drive the final adsorption capacity and glucose yields. Quantitatively understanding of interaction mechanism between lignin and cellulases is essential for the efficient improvement of lignocellulose enzymatic hydrolysis. However, the individual contribution of multiple forces between lignin and cellulases to the non-productive adsorption of enzymes still remains deeply ambiguous, especially in situations of near enzymatic hydrolysis temperatures. Herein, atomic force microscopy (AFM) and computational simulations were utilized to quantitatively analyze the intermolecular forces between lignin and enzyme at 25 °C and 40 °C. Our results unveiled that an increase in temperature obviously improved adsorption capacity and total intermolecular forces between lignin and cellulases. This positive relationship mainly comes from the increase in the decay length of hydrophobic forces for lignin-cellulases when temperature increases. Different from the hydrophobic interaction which provides long-range part of attractions, van der Waals forces dominate the intermolecular force only at approaches < 2 nm. On the other hand, electrostatic forces exhibited repulsive effects, and its intensity and distance were limited due to the low surface potential of cellulases. Short-range forces including hydrogen bonding (main) and π-π stacking (minor) stabilize the non-specific binding of enzymes to lignin, but increasing temperature reduces hydrogen bond number. Therefore, the relative contribution of long-range forces increased markedly at higher temperatures, which benefits protein capture and brings lignin and cellulase close together. Finally, the structure–activity relationships between lignin physicochemical properties and its inhibitory effect to enzymes indicated that hydrophobic interactions, hydrogen bonding, and steric effects drive the final adsorption capacity and glucose yields. This work provides quantitative and basic insights into the mechanism of lignin-cellulase interfacial interactions and guides design of saccharification enhancement approaches. [ABSTRACT FROM AUTHOR]

Effects of nonionic surfactants on enzymatic hydrolysis of Avicel at different agitation rates and solid loadings and the mechanism were studied. Nonionic surfactants couldn’t improve the enzymatic hydrolysis efficiency at 0 and 100 rpm but could enhance the enzymatic hydrolysis significantly at high agitation rate (200 and 250 rpm). Cellulase was easily deactivated at high agitation rate and the addition of nonionic surfactants can protect against the shear-induced deactivation, especially when the cellulase concentration was low. When 25 mg protein/L of cellulase solution was incubated at 200 rpm for 72 h, the enzyme activity increased from 36.0% to 89.5% by adding PEG4600. Moreover nonionic surfactants can compete with enzyme in air-liquid interface and reduce the amount of enzyme exposed in the air-liquid interface. The mechanism was proposed that nonionic surfactants could enhance the enzymatic hydrolysis of Avicel by reducing the cellulase deactivation caused by shear force and air-liquid interface. [ABSTRACT FROM AUTHOR]

Polyvinylpyrrolidone (PVP) is an antifouling polymer to resist the adsorption of protein on solid surface. Effects of PVP on the enzymatic hydrolysis of pretreated lignocelluloses and its mechanism were studied. Adding 1 g/L of PVP8000, the enzymatic digestibility of eucalyptus pretreated by dilute acid (Eu-DA) was increased from 28.9% to 73.4%, which is stronger than the classic additives, such as PEG, Tween and bovine serum albumin. Compared with PEG4600, the adsorption of PVP8000 on lignin was larger, and the adsorption layer was more stable and hydrophilic. Therefore, PVP8000 reduced 73.1% of the cellulase non-productive adsorption on lignin and enhanced the enzymatic hydrolysis of lignocelluloses greatly. [ABSTRACT FROM AUTHOR]

Two pretreatment methods to overcome recalcitrance of lignocelluloses, sulfite pretreatment (SPORL) and dilute acid (DA), were conducted to pretreat softwood masson pine and hardwood eucalyptus for enzymatic hydrolysis. In the presence of corresponding pre-hydrolysates, adding moderate cetyltrimethylammonium bromide (CTAB) could enhance the enzymatic hydrolysis of the SPORL-pretreated substrates, but had no enhancement for the DA-pretreated substrates. The results showed that sodium lignosulfonate (SL) in pre-hydrolysates and CTAB together had a strong enhancement on the enzymatic hydrolysis of lignocelluloses. The compound of commercial lignosulfonate SXSL and CTAB (SXSL-CTAB) could enhance the substrate enzymatic digestibility (SED) of SPORL-pretreated masson pine from 27.1% to 71.0%, and that of DA-pretreated eucalyptus from 37.6% to 67.9%. The mechanism that CTAB increased the adsorption of SL on lignin to form more effective steric hindrance and reduced the non-productive adsorption of cellulase on lignin by neutralizing the negative charge of SL was proposed. [ABSTRACT FROM AUTHOR]

Effect of the molecular structure of lignosulfonate-based polyoxyethylene ether (LS-PEG) on the enzymatic hydrolysis of Avicel and corn stover was investigated. With the increase of PEG contents and M w of LS-PEG, glucose yield of corn stover increased from 16.7% to 51.9%. When LS-PEG was compounded with cationic surfactant cetyltrimethylammonium bromide (CTAB), the enhancement of glucose yield of corn stover was further increased from 45.3% to 62.8%. Sulfonic group of LS-PEG preferentially interacted with quaternary ammonium group of CTAB by the electrostatic attraction to form a similar non-ionic surfactant CTAB-LS-PEG. CTAB-LS-PEG showed electrically neutral and more hydrophobic, and blocked more nonproductive adsorption of cellulase on the lignin. Therefore, CTAB-LS-PEG enhanced enzymatic hydrolysis efficiency of lignocelluloses more significantly. The understanding developed in this study can help to develop potential approaches and strategies for effective application of lignosulfonate to improve bioconversion of lignocellulosic biomass. [ABSTRACT FROM AUTHOR]

Sodium lignosulfonate (SXSL) and long-chain fatty alcohols (LFAs) could enhance the enzymatic hydrolysis of xylan, and the compound of SXSL and LFAs have synergies on the enzymatic hydrolysis. SXSL shows a strong enhancement in buffer pH range from 4.0 to 6.0. The enhancement increased with the SXSL dosage and the xylanase loading. The cellulose and lignin in corncob substrate could not only adsorb xylanase nonproductively, but also seriously reduce the accessibility of xylanase on xylan to impede the enzymatic hydrolysis of xylan. Cellulase could break the plant cell wall structure of corncob and make additives work better. The xylose yield of corncob at 72 h increased from 59.4% to 73.7% by adding the compound of 5 g/L SXSL and 0.01% (v/v) n -decanol, which was higher than that without cellulase and additives by 30.7%. Meanwhile, the glucose yield at 72 h of corncob increased from 45.8% to 62.3%. [ABSTRACT FROM AUTHOR]

• Highly recyclable SC was used in lignocellulosic enzymatic hydrolysis. • Using SC to enhance cellulase recovery through ingenious pH regulation. • SC increased the sugar yield of Eu-DA by 95.5% and save at least 40% cellulase. Most additives that capable of enhancing enzymatic hydrolysis of lignocellulose are petroleum-based, which are not easy to recycle with poor biodegradability. In this work, highly recyclable and biodegradable sodium caseinate (SC) was used to enhance lignocellulosic hydrolysis with improved cellulase recyclability. When the pH decreased from 5.5 to 4.8, more than 96% SC could be precipitated from the solution and recovered. Adding SC increased enzymatic digestibility of dilute acid pretreated eucalyptus (Eu-DA) from 39.5% to 78.2% under Eu-DA loading of 10 wt% and pH = 5.5, and increase cellulase content in 72 h hydrolysate from only 15.2% of the original to 60.0%, which facilitated the recovery of cellulases through re-adsorption by fresh substrates. With multiple cycles of re-adsorption, application of SC not only increased the sugar yield of Eu-DA by 95.5%, but also reduced cellulase loading by 40%. [ABSTRACT FROM AUTHOR]