Comparison of Alkaline Sulfite Pretreatment and Acid Sulfite Pretreatment with Low Chemical Loading in Saccharification of Poplar.

Sulfite pretreatment is a productive process for lignin dissolution in lignocelluloses and to reduce the hydrophobicity of lignin by sulfonation, thus promoting the hydrolyzability of the substrate. Previously, sulfite pretreatment needs high dosages of chemicals and thus results in the high cost of the pretreatment and the great pressure of environmental pollution. To overcome these problems, it was crucial to research whether alkaline sulfite pretreatment (ALS) and acid sulfite pretreatment (ACS) with low chemical loading could enhance the saccharification of poplar. In this work, the results indicated that with low loading of chemicals in sulfite pretreatment, ALS pretreatment (1.6% Na2SO3 and 0.5% NaOH) at 180 °C removed more lignin, resulted in lower hydrophobicity and higher cellulase adsorption capacity of poplar than ACS pretreatment (1.6% Na2SO3 and 0.5% H2SO4) at 180 °C. A satisfying glucose yield of 84.9% and a xylose yield of 76.0% were obtained from poplar after ALS pretreatment with 1.6% Na2SO3 and 0.5% NaOH at 180 °C for 1 h using 10 FPU cellulase/g dry matter, saving sodium sulfite by 60.0% compared to the loading of sulfite in traditional sulfite pretreatment. The strategy developed in this work reduced chemical loading and cellulase loading in alkali sulfite pretreatment for the saccharification of poplar.

Improving cellulases hydrolytic action: An expanded role for electron donors of lytic polysaccharide monooxygenases in cellulose saccharification.

Ascorbic acid (AscA) and gallic acid (GalA) are common electron donors and their boosting effect on lytic polysaccharide monooxygenases (LPMO) has been studied extensively. However, their influence on cellulase hydrolytic action has been ignored. In this work, the effect of AscA and GalA on cellulases hydrolytic action was evaluated. It was found that AscA could increase the hydrolysis of cellulose by cellulases, while GalA showed no effect on cellulases' hydrolytic action. The effect of AscA differed for the monocomponent cellulases: it showed a special boosting effect on cellobiohydrolase, rather than endoglucanase and ß-glucosidase. This promoting effect could be another mechanism behind the boosting effect of the AscA-driven LPMO system on cellulose saccharification. These findings thus advance the understanding of the role of electron donors on cellulose saccharification and offer important clues on how to evaluate the feasibility of electron donors from a new perspective.

Structural properties and hydrolysabilities of Chinese Pennisetum and Hybrid Pennisetum: Effect of aqueous ammonia pretreatment.

The effects of aqueous ammonia pretreatment on structural properties and hydrolysabilities of Chinese Pennisetum and Hybrid Pennisetum were investigated. Aqueous ammonia pretreatment increased cellulose crystallinities and hydrolysabilities of Chinese Pennisetum and Hybrid Pennisetum. Compared with Chinese Pennisetum, Hybrid Pennisetum showed better enzymatic digestibility. Xylanase supplementation was more effective than the increase of cellulase loadings in the hydrolysis of aqueous ammonia pretreated Chinese Pennisetum and Hybrid Pennisetum. After supplementation of 2mg of xylanase/g dry matter to 5 FPU of cellulases/g dry matter, the hydrolysis yields of cellulose of aqueous ammonia pretreated Chinese Pennisetum and Hybrid Pennisetum were 92.3-95.4%, and the hydrolysis yields of xylan were 86.9-94.2%. High hydrolysability and low dosage of enzyme loadings together with the advantages of high yield and widely distribution demonstrated the potential of Chinese Pennisetum and Hybrid Pennisetum for the production of platform sugars.

Comparison of aqueous ammonia and dilute acid pretreatment of bamboo fractions: Structure properties and enzymatic hydrolysis.

The effect of two pretreatments methods, aqueous ammonia (SAA) and dilute acid (DA), on the chemical compositions, cellulose crystallinity, morphologic change, and enzymatic hydrolysis of bamboo fractions (bamboo yellow, timber, green, and knot) was compared. Bamboo fractions with SAA pretreatment had better hydrolysability than those with DA pretreatment. High crystallinity index resulted in low hydrolysis yield in the conversion of SAA pretreated bamboo fractions, not DA pretreated fractions. The increase of cellulase loading had modestly positive effect in the hydrolysis of both SAA and DA pretreated bamboo fractions, while supplement of xylanase significantly increased the hydrolysis of the pretreated bamboo fractions, especially after SAA pretreatment. The results indicated that SAA pretreatment was more effective than DA pretreatment in conversion of bamboo fractions, and supplementation of xylanase was necessary in effective conversion of the SAA pretreated fractions into fermentable sugars.

Competitive inhibition of cellobiohydrolase I by manno-oligosaccharides.

In the hydrolysis of softwood, significant amounts of manno-oligosaccharides (MOS) are released from mannan, the major hemicelluloses in softwood. However, the impact of MOS on the performance of cellulases is not yet clear. In this work, the effect of mannan and MOS in cellulose hydrolysis by cellulases, especially cellobiohydrolase I (CBHI) from Thermoascus aurantiacus (Ta Cel7A), was studied. The glucose yield of Avicel decreased with an increasing amount of added mannan. Commercial cellulases contained mannan hydrolysing enzymes, and ß-glucosidase played an important role in mannan hydrolysis. Addition of 10mg/ml mannan reduced the glucose yield of Avicel (at 20g/l) from 40.1 to 24.3%. No inhibition of ß-glucosidase by mannan was observed. The negative effects of mannan and MOS on the hydrolytic action of cellulases indicated that the inhibitory effect was at least partly attributed to the inhibition of Ta Cel7A (CBHI), but not on ß-glucosidase. Kinetic experiments showed that MOS were competitive inhibitors of the CBHI from T. aurantiacus, and mannobiose had a stronger inhibitory effect on CBHI than mannotriose or mannotetraose. For efficient hydrolysis of softwood, it was necessary to add supplementary enzymes to hydrolyze both mannan and MOS to less inhibitory product, mannose.

Enhanced xylanase performance in the hydrolysis of lignocellulosic materials by surfactants and non-catalytic protein.

Addition of additives has been confirmed to increase cellulase performance in the hydrolysis of lignocellulosic materials. In the hydrolysis of xylan-containing lignocellulosic biomass, xylanase can synergistically enhance the performance of cellulase. However, the role of additives in xylan hydrolysis by xylanase is not yet clear. In this work, with the presence of additives (bovine serum albumin, poly(ethylene glycol), and Tween), the hydrolysis of isolated xylan and the xylan in corn stover increased to different extents. Additives increased free xylanase in supernatants in the hydrolysis with xylanase, indicating the reduction of the adsorption of xylanase on corn stover and insoluble xylan. Enhanced hydrolysis of Avicel and corn stover by additives suggested that besides the prevention of unproductive binding of xylanase to lignin by additives, reducing the adsorption of xylanase on substrates was also contributed to enzymatic hydrolysis. The increment of xylanase activity by additives suggests that the additives were activators of xylanase. The results of this work indicate that the supplementation of additives could improve xylanase performance, synergistically enhanced the cellulose hydrolysis, and beneficial for the recycling of xylanase.

Hydrolyzabilities of different corn stover fractions after aqueous ammonia pretreatment.

The effect of aqueous ammonia pretreatment on the hydrolysis of different corn stover fractions (rind, husk, leaf, and pith) by xylanase (XYL) with cellulases (CELs) was evaluated. The aqueous ammonia pretreatment had excellent delignification ability (above 66%) for different corn stover fractions. The corn rind exhibited the lowest susceptibility to aqueous ammonia pretreatment. The pretreated rind showed the lowest hydrolyzability by CEL and XYL, which was supported by a high content of crystalline cellulose in the hydrolyzed residues of rind, as confirmed by X-ray diffraction (XRD). With the addition of 1 mg XYL/g dry matter, a high glucose yield (above 90%) could be obtained from the pretreated rind by CEL. The results revealed that a high hydrolyzate yield of corn rind after aqueous ammonia pretreatment could be obtained with 1 mg xylanase/g dry matter, showing that aqueous ammonia pretreatment and xylanase addition to cellulases have great potential for the efficient hydrolysis of corn stover without previous fractionation.

Behavior of cellulose and xylan in aqueous ammonia pretreatment.

The effect of aqueous ammonia on the solubilization of cellulose and xylans was investigated by detecting the amounts of reducing sugars and monosaccharides in the treatment liquors. The degree of cellulose and xylan solubilization increased with the increase of treatment temperature. When the treatment temperature increased from 20 to 90 °C, the amounts of reducing sugars released from Avicel and cellulose fiber by 21 % ammonia at a solid to liquid ratio of 1:10 for 24 h increased from 1.0 and 0.9 to 4.4 and 2.7 mg/g dry matter (DM), respectively. The amounts of reducing sugars released from wheat straw, beechwood, and oat spelt xylans increased from 1.2-7.0 to 3.3-13.5 mg/g DM. Xylans appeared to be more susceptible than cellulose in aqueous ammonia treatment. Structure analysis of untreated and treated Avicel and cellulose fiber showed that aqueous ammonia increased the specific surface area and crystallinity index of cellulose. Most of the cellulose and xylan that were solubilized existed in the form of oligomers such as cello-oligosaccharides and xylo-oligosaccharides. Xylobiose and xylotriose were the main oligosaccharides released from oat spelt xylan by aqueous ammonia treatment as confirmed by electrospray ionization-mass spectrometry. The results here indicated that a slight amount of cellulose and xylans was solubilized and low amounts of cellulase inhibitors, oligomers, were found during mild aqueous ammonia pretreatment process. Therefore, from the economical perspectives, mild ammonia pretreatment would be favorable for aqueous ammonia pretreatment of lignocelluloses.