Shiyuan Yu

Three-stage hydrolysis to enhance enzymatic saccharification of steam-exploded corn stover.

The objective of the present research was to explore new approach to reduce the hydrolysis time and to enhance the productivity of enzymatic saccharification. One-stage hydrolysis of steam-exploded corn stover required 72 h to reach a yield of 62.8%, while multi-stage hydrolysis could reduce the time to 24 h. A concept of three-stage hydrolysis was therefore proposed in which cellulosic substrate was hydrolyzed for 6, 6, and 12 h, respectively. High hydrolysis yields, 70.2% with enzyme recycling and 76.1% with the supplement of fresh enzyme to eliminate enzyme recovery procedure, were obtained in 24 h. Analysis indicated that short-time hydrolysis and the removal of end products at each stage improved cellulase activities and benefited the adsorption of cellulase enzyme to the solid substrate. When steam-exploded corn stover was used as the substrate for cellulase synthesis, a hydrolysis yield of 88.6% was achieved in 24 h.

Optimization of culture conditions for production of yeast biomass using bamboo wastewater by response surface methodology

Response surface methodology (RSM) was applied to optimize culture conditions for the growth of Candida utilis with bamboo wastewater. A significant influence of initial pH, fermentation time and yeast extract on biomass of C. utilis was evaluated by Plackett-Burman design (PBD). These factors were further optimized using a central composite design (CCD) and RSM. A combination of initial pH 6.1, fermentation time 69 h and yeast extract 1.17 g/L was optimum for maximum biomass of C. utilis. A 1.7-fold enhancement of biomass of C. utilis was gained after optimization in shake-flask cultivation. The biomass of C. utilis reached 19.17 g/L in 3 L fermentor.

Detoxification of corn stover prehydrolyzate by trialkylamine extraction to improve the ethanol production with Pichia stipitis CBS 5776

In order to realize the separated ethanol fermentation of glucose and xylose, prehydrolysis of corn stover with sulfuric acid at moderate temperature was applied, while inhibitors were produced inevitably. A complex extraction was adopted to detoxify the prehydrolyzate before fermentation to ethanol with Pichia stipitis CBS 5776. The best proportion of mixed extractant was 30% trialkylamine-50% n-octanol -20% kerosene. Detoxification results indicated that 73.3% of acetic acid, 45.7% of 5-hydroxymethylfurfural and 100% of furfural could be removed. Compared with the undetoxified prehydrolyzate, the fermentability of the detoxified prehydrolyzate was significantly improved. After 48 h fermentation of the detoxified prehydrolyzate containing 7.80 g/l of glucose and 52.8 g/l of xylose, the sugar utilization ratio was 93.2%; the ethanol concentration reached its peak value of 21.8 g/l, which was corresponding to 82.3% of the theoretical value.

Three-stage enzymatic hydrolysis of steam-exploded corn stover at high substrate concentration.

The feasibility of three-stage hydrolysis of steam-exploded corn stover at high-substrate concentration was investigated. When substrate concentration was 30% and enzyme loading was 15-30 FPU/g cellulose, three-stage (9+9+12 h) hydrolysis could reach a hydrolysis yield of 59.9-81.4% in 30 h. Compared with one-stage hydrolysis for 72 h, an increase of 34-37% in hydrolysis yield could be achieved. When steam-exploded corn stover was used as the substrate for enzyme synthesis and hydrolysis was conducted at a substrate concentration of 25% with an enzyme loading of 20 FPU/g cellulose, a hydrolysis yield of 85.1% was obtained, 19% higher than that the commercial cellulase could reach under the same conditions. The removal of end products was suggested to improve the adsorption of cellulase on the substrate and enhance the productivity of enzymatic hydrolysis.

Remarkable solvent and extractable lignin effects on enzymatic digestibility of organosolv pretreated hardwood.

Low solvent concentration effect on substrate digestibility of ethanol organosolv pretreated sweetgum was examined. Surprisingly, lower ethanol concentration in organosolv pretreatments resulted in faster initial rates and higher 72h hydrolysis yields in pretreated substrates. A strong correlation (r(2)=0.96) between pretreatment combined severity factor and residual xylan/glucan ratio was observed. The residual xylan/glucan ratio was associated with the initial hydrolysis rate closely. Furthermore, it was found that preserving extractable lignin in the pretreated substrates could improve enzymatic hydrolysis yield by 33%. This has an important implication in reducing the pretreatment and enzyme cost, because the typical solvent washing after pretreatment could be eliminated and preserving extractable lignin could reduce enzyme loading. Finally, we observed that xylan removal by xylanase could improve the initial rate by 53% and increase the 72h hydrolysis yield by 21%. The extractable lignin precipitation on pretreated substrates increased the 72h hydrolysis yield by 10%.

Contrasting effects of hardwood and softwood organosolv lignins on enzymatic hydrolysis of lignocellulose.

Identifying an appropriate parameter to elucidate effects of lignin on enzymatic hydrolysis is essential to understand the interactions between enzymes and lignin. Contrasting effects of hardwood organosolv lignin (EOL-SG) and softwood organosolv lignin (EOL-LP) on enzymatic hydrolysis were observed. The addition of EOL-SG (8 g/L) significantly improved the 72 h hydrolysis yields of organosolv pretreated sweetgum (OPSG) and loblolly pine (OPLP) from 49.3% to 68.6% and from 41.2% to 60.8%, respectively. In contrast, the addition of EOL-LP decreased the 72 h hydrolysis yields of OPSG and OPLP to 42.0% and 38.1%, respectively. A strong correlation between the distribution coefficients of cellulase enzymes on lignins and the changes of hydrolysis yields indicated that the inhibitory or stimulatory effects of organosolv lignins on enzymatic hydrolysis were governed by the distribution coefficients (R). The different R values probably were related to the electrostatic interactions, hydrophobic interactions and hydrogen bondings between enzymes and lignin.

Impacts of lignocellulose-derived inhibitors on L-lactic acid fermentation by Rhizopus oryzae.

Inhibitors generated in the pretreatment and hydrolysis of corn stover and corn cob were identified. In general, they inhibited cell growth, lactate dehydrogenase, and lactic acid production but with less or no adverse effect on alcohol dehydrogenase and ethanol production in batch fermentation by Rhizopus oryzae. Furfural and 5-hydroxymethyl furfural (HMF) were highly toxic at 0.5-1 g L(-1), while formic and acetic acids at less than 4 g L(-1) and levulinic acid at 10 g L(-1) were not toxic. Among the phenolic compounds at 1 g L(-1), trans-cinnamic acid and syringaldehyde had the highest toxicity while syringic, ferulic and p-coumaric acids were not toxic. Although these inhibitors were present at concentrations much lower than their separately identified toxic levels, lactic acid fermentation with the hydrolysates showed much inferior performance compared to the control without inhibitor, suggesting synergistic or compounded effects of the lignocellulose-degraded compounds on inhibiting lactic acid fermentation.

An integrated process to produce ethanol, vanillin, and xylooligosaccharides from Camellia oleifera shell

This study aims to present an integrated process that can be used to produce ethanol, vanillin, and xylooligosaccharides from Camellia oleifera shell. After the shell was pretreated with NaOH, two fractions were obtained: solid and liquid fractions. The solid fraction was hydrolyzed with cellulase and then fermented with Pichia stipitis to produce ethanol. The liquid fraction was subjected to oxidation to prepare vanillin or hydrolysis with xylanase to prepare xylooligosaccharides. The optimal pretreatment conditions of an orthogonal test were as follows: 12% NaOH concentration; 120°C; 150 min; and liquid-solid ratio of 10.0. After pretreatment, the solid fraction containing cellulose and a small part of xylan at 10% substance concentration via enzymatic hydrolysis and glucose-xylose cofermentation could obtain 17.35 g/L of ethanol, 80.90% of the theoretical yield. The liquid fraction was initially hydrolyzed with xylanase to produce 1758.63 mg/L of xylooligosaccharides (DP2-6) and then oxidized to produce 322.07 mg/L of vanillin.

Co-production of functional xylooligosaccharides and fermentable sugars from corncob with effective acetic acid prehydrolysis

A novel and green approach for the coproduction of xylooligosaccharides (XOS), in terms of a series of oligosaccharide components from xylobiose to xylohexose, and fermentable sugars was developed using the prehydrolysis of acetic acid that was fully recyclable and environmentally friendly, followed by enzymatic hydrolysis. Compared to hydrochloric acid and sulfuric acid, acetic acid hydrolysis provided the highest XOS yield of 45.91% and the highest enzymatic hydrolysis yield. More than 91% conversion of cellulose was achieved in a batch-hydrolysis using only a cellulase loading of 20FPU/g cellulose and even a high solid loading of 20% without any special strategies. The acetic acid pretreated corncob should be washed adequately before saccharification to achieve complete hydrolysis. Consequently, a mass balance analysis showed that 139.8g XOS, 328.1g glucose, 25.1g cellobiose, and 147.8g xylose were produced from 1000g oven dried raw corncob.

Disparate roles of solvent extractable lignin and residual bulk lignin in enzymatic hydrolysis of pretreated sweetgum

The roles of solvent extractable lignin and residual bulk lignin in enzymatic hydrolysis of Avicel and lignocellulosic biomass were distinguished in this study. Solvent extractable lignin removal reduced the 72 h hydrolysis yields of dilute acid pretreated sweetgum (DASG) and organosolv pretreated sweetgum (OPSG) from 38.1% to 31.8% and from 69.9 to 49.3%, respectively. On the contrary, residual bulk lignin removal enhanced the 72 h hydrolysis yields of DASG and OPSG to 91.7% and 90.5%, respectively. The isolated lignins were added into enzymatic hydrolysis of Avicel, which revealed the positive effect of extractable lignin and the negative effect of residual bulk lignin on enzymatic hydrolysis. The cellulase distribution during the hydrolysis and cellulase adsorption indicated that the extractable lignin could counter the negative effect of residual bulk lignin by reducing the non-productive binding between cellulase and bulk lignin.