Adsorption characteristics of cellulase and β-glucosidase on Avicel, pretreated sugarcane bagasse, and lignin.

Although adsorption is an essential step in the enzymatic hydrolysis of lignocellulosic materials, literature reports controversial results in relation to the adsorption of the cellulolitic enzymes on different biomasses/pretreatments, which makes difficult the description of this phenomenon in hydrolysis mathematical models. In this work, the adsorption of these enzymes on Avicel and sugarcane bagasse pretreated by the hydrothermal bagasse (HB) and organosolv bagasse (OB) methods was evaluated. The results have shown no significant adsorption of β-glucosidase on Avicel or HB. Increasing solids concentration from 5% (w/v) to 10% (w/v) had no impact on the adsorption of cellulase on the different biomasses if stirring rates were high enough (>100 rpm for Avicel and >150 rpm for HB and OB). Adsorption equilibrium time was low for Avicel (10 Min) when compared with the lignocellulosic materials (120 Min). Adsorption isotherms determined at 4 and 50 °C have shown that for Avicel there was a decrease in the maximum adsorption capacity (Emax) with the temperature increase, whereas for HB increasing temperature increased Emax . Also, Emax increased with the content of lignin in the material. Adsorption studies of cellulase on lignin left after enzymatic digestion of HB show lower but significant adsorption capacity (Emax = 11.92 ± 0.76 mg/g).

Machado DL ，Moreira Neto J ，da Cruz Pradella JG ，Bonomi A ，Rabelo SC ，da Costa AC ... -  《-》

Access of cellulase to cellulose and lignin for poplar solids produced by leading pretreatment technologies.

Adsorption of cellulase on solids resulting from pretreatment of poplar wood by ammonia fiber expansion (AFEX), ammonia recycled percolation (ARP), controlled pH, dilute acid (DA), flowthrough (FT), lime, and sulfur dioxide (SO(2)) and pure Avicel glucan was measured at 4 degrees C, as were adsorption and desorption of cellulase and adsorption of beta-glucosidase for lignin left after enzymatic digestion of the solids from these pretreatments. From this, Langmuir adsorption parameters, cellulose accessibility to cellulase, and the effectiveness of cellulase adsorbed on poplar solids were estimated, and the effect of delignification on cellulase effectiveness was determined. Furthermore, Avicel hydrolysis inhibition by enzymatic and acid lignin of poplar solids was studied. Flowthrough pretreated solids showed the highest maximum cellulase adsorption capacity (sigma(solids) = 195 mg/g solid) followed by dilute acid (sigma(solids) = 170.0 mg/g solid) and lime pretreated solids (sigma(solids) = 150.8 mg/g solid), whereas controlled pH pretreated solids had the lowest (sigma(solids) = 56 mg/g solid). Lime pretreated solids also had the highest cellulose accessibility (sigma(cellulose) = 241 mg/g cellulose) followed by FT and DA. AFEX lignin had the lowest cellulase adsorption capacity (sigma(lignin) = 57 mg/g lignin) followed by dilute acid lignin (sigma(lignin) = 74 mg/g lignin). AFEX lignin also had the lowest beta-glucosidase capacity (sigma(lignin) = 66.6 mg/g lignin), while lignin from SO(2) (sigma(lignin) = 320 mg/g lignin) followed by dilute acid had the highest (301 mg/g lignin). Furthermore, SO(2) followed by dilute acid pretreated solids gave the highest cellulase effectiveness, but delignification enhanced cellulase effectiveness more for high pH than low pH pretreatments, suggesting that lignin impedes access of enzymes to xylan more than to glucan, which in turn affects glucan accessibility. In addition, lignin from enzymatic digestion of AFEX and dilute acid pretreated solids inhibited Avicel hydrolysis less than ARP and flowthrough lignin, whereas acid lignin from unpretreated poplar inhibited enzymes the most. Irreversible binding of cellulase to lignin varied with pretreatment type and desorption method.

Kumar R ，Wyman CE 《-》

Lignin prepared from different alkaline pretreated sugarcane bagasse and its effect on enzymatic hydrolysis.

Sugarcane bagasse (SCB) was pretreated by sodium hydroxide (SH), alkaline ethanol (AE) and alkaline hydrogen peroxide (AHP), and the solid residue with similar lignin content after different pretreatment was selected. Enzymatic saccharification with different substrates was performed at 2%, 10%, and 20% (w/v) solid loading. After that, the lignin from different substrate was extracted and its structure was characterized. Furthermore, the adsorption capability of isolated lignin to cellulase and its effect on enzymatic hydrolysis were studied. The results showed that, as the substrate content increased from 2% (w/v) to 20% (w/v), the glucose yield after digestion of SH, AE, and AHP pretreated SCB reduced by 41.8%, 35.4%, and 28.7%, respectively. The inhibitory effect of different prepared lignin on the digestibility of Avicel was as follows, SH pretreated lignin (SHL) > cellulase enzymatic hydrolysis lignin (CEL) > AE pretreated lignin (AEL) > AHP pretreated lignin (AHPL), which exhibited positive correlation with its non-productive adsorption capability to cellulase. Lignin samples with low negative zeta potential, high hydrophobicity and high ration of syringyl (S) to guaiacyl (G) were unfavorable for the enzymatic saccharification of cellulose.

Xu C ，Zhang J ，Zhang Y ，Guo Y ，Xu H ，Liang C ，Wang Z ，Xu J ... -  《-》

Temperature dependent cellulase adsorption on lignin from sugarcane bagasse.

Extents of adsorption of cellulolytic enzymes on lignin, derived from sugarcane bagasse, were an inverse function of incubation temperature and varied with type of lignin extraction. At 45 °C, lignin derived from acid hydrolyzed liquid hot water pretreated bagasse completely adsorbed cellulolytic enzymes from Trichoderma reesei within 90 min. Lignin derived from enzyme hydrolyzed liquid hot water pretreated bagasse adsorbed only 60% of T. reesei endoglucanase, exoglucanase and β-glucosidase activities. β-Glucosidase from Aspergillus niger was not adsorbed. At 30 °C, adsorption of all of the enzymes was minimal and enzyme hydrolysis at 30 °C approached that at 45 °C after 168 h. Hence, temperature provided an approach to decrease loss of enzyme activity by reducing enzyme adsorption on lignin. This helps to explain why simultaneous saccharification and fermentation (SSF) and consolidated bioprocessing (CBP), both carried out at 30-32 °C, could offer viable options for mitigating lignin-derived inhibition effects.

Zanchetta A ，Dos Santos ACF ，Ximenes E ，da Costa Carreira Nunes C ，Boscolo M ，Gomes E ，Ladisch MR ... -  《-》

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 (40 mg lignin/200 mg Avicel) decreased by 10.74%, 9.28%, 8.73%, 4.22%, and 2.80% after digestion of Avicel for 72 h with the presence of EL, SHL, CEL, HLWL, and DSAL, respectively.

Xu C ，Liu F ，Alam MA ，Chen H ，Zhang Y ，Liang C ，Xu H ，Huang S ，Xu J ，Wang Z ... -  《-》