Combined oxidization and liquid ammonia pretreatment of bamboo of various ages and species for maximizing fermentable sugar release.

To determine the potential for improving biomass enzymolysis, a combined oxidization and liquid ammonia pretreatment (OD-LAT) was employed for bamboo. The effects of oxidant, bamboo ages, and species on the pretreatment effectiveness and subsequent enzymolysis were studied. Under the optimal OD-LAT pretreatment and enzymolysis of the B-NA bamboo Neosinocalamus affinis, the glucan and xylan conversion reached 83.85% and 78.66%, respectively, and approximately 59.7-68.5 g of fermentable sugars can be produced per 100 g of dry bamboo, which was an approximately 5-8 fold increase compared with untreated sample. The H2O2 loading of 1.0 was the optimal oxidant dosage for the OD-LAT process. The OD-LAT pretreatment was only suitable for bamboo under three-year-old, and it significantly improved the enzymolysis of B-NA and B-BM, while it was limited to B-DO and B-PP. The pretreatment effects of bamboo were not only related to composition but also to the bamboo age, species, macro-structures and micro-structures.

Recent advances on ammonia-based pretreatments of lignocellulosic biomass.

Ammonia-based pretreatments have been extensively studied in the last decade as one of the leading pretreatment technologies for lignocellulose biorefining. Here, we discuss the key features and compare performances of several leading ammonia-based pretreatments (e.g., soaking in aqueous ammonia or SAA, ammonia recycled percolation or ARP, ammonia fiber expansion or AFEX, and extractive ammonia or EA). We provide detailed insight into the distinct physicochemical mechanisms employed during ammonia-based pretreatments and its impact on downstream bioprocesses (e.g., enzymatic saccharification); such as modification of cellulose crystallinity, lignin/hemicellulose structure, and other ultrastructural changes such as cell wall porosity. Lastly, a brief overview of process technoeconomics and environmental impacts are discussed, along with recommendations for future areas of research on ammonia-based pretreatments.

Production of glycoprotein bioflocculant from untreated rice straw by a CAZyme-rich bacterium, Pseudomonas sp. HP2.

It has been reported that certain biomass-degrading bacteria can produce bioflocculant through directly utilizing untreated biomass as carbon source. However, little is known about the synthesis mechanism of bioflocculant in these bacteria. In this study, a biomass-degrading bacterium Pseudomonas sp. HP2 showing excellent production ability of bioflocculant was isolated from the forest soil. The HP2 strain secreted alkali-thermo-tolerant CMCase and xylanase, with the maximum activities of 0.06 and 1.07 U ml-1, respectively, when the untreated rice straw was used as carbon source. The maximum flocculating efficiency with the value of 92.5% was produced from untreated rice straw by HP2 strain. Component analysis showed that this bioflocculant was abundant in the amino acids and monosaccharides with the total contents of 384.9 and 478.3 mg g-1 dry bioflocculant, respectively. The most amino acid and monosaccharide in this bioflocculant were proline and rhamnose, which accounted for 26.5% and 33.3% of total amino acids and total monosaccharides, respectively. To explore the synthesis mechanism of bioflocculant in HP2, the genome of HP2 strain was measured by Illumina HiSeq PE150 platform. The results showed that the genome of HP2 strain possessed abundant CAZy family related genes, which may play an important role in biomass degradation and bioflocculant synthesis.

Pretreatment of Miscanthus with biomass-degrading bacteria for increasing delignification and enzymatic hydrolysability.

Biomass recalcitrance is still a main challenge for the production of biofuels and high-value products. Here, an alternative Miscanthus pretreatment method by using lignin-degrading bacteria was developed. Six efficient Miscanthus-degrading bacteria were first cultured to produce laccase by using 0.5% Miscanthus biomass as carbon source. After 1-5 days of incubation, the maximum laccase activities induced by Miscanthus in the six strains were ranged from 103 to 8091 U l-1 . Then, the crude enzymes were directly diluted by equal volumes of citrate buffer and added Miscanthus biomass to a solid concentration at 4% (w/v). The results showed that all bacterial pretreatments significantly decreased the lignin content, especially in the presence of two laccase mediators (ABTS and HBT). The lignin removal directly correlated with increases in total sugar and glucose yields after enzymatic hydrolysis. When ABTS was used as a mediator, the best lignin-degrading bacteria (Pseudomonas sp. AS1) can remove up to 50.1% lignin of Miscanthus by obtaining 2.2-fold glucose yield, compared with that of untreated biomass. Therefore, this study provided an effective Miscanthus pretreatment method by using lignin-degrading bacteria, which may be potentially used in improving enzymatic hydrolysability of biomass.

The use of steam explosion to increase the nutrition available from rice straw.

In the present study, rice straw was pretreated using steam-explosion (ST) technique to improve the enzymatic hydrolysis of potential reducing sugars for feed utilization. The response surface methodology based on central composite design was used to optimize the effects of steam pressure, pressure retention time, and straw moisture content on the yield of reducing sugar. All the investigated variables had significant effects (P < 0.001) on the reducing sugar yield. The optimum yield of 30.86% was obtained under the following pretreatment conditions: steam pressure, 1.54 MPa; pressure retention time, 140.5 Sec; and straw moisture content, 41.6%. The yield after thermal treatment under the same conditions was approximately 16%. Infrared (IR) radiation analysis showed a decrease in the cellulose IR crystallization index. ST noticeably increases reducing sugars in rice straw, and this technique may also be applicable to other cellulose/lignin sources of biomass.

Effects of compositional changes of AFEX-treated and H-AFEX-treated corn stover on enzymatic digestibility.

Corn stover is one of the main agricultural residues being considered as a cellulosic ethanol feedstock. This work evaluated the effectiveness of AFEX™(1) pretreatment for converting corn stover to fermentable sugars, both with and without pre-soaking in hydrogen peroxide. The compositional changes and enzymatic digestibility of AFEX-treated and H-AFEX-treated biomass were investigated. Results showed that most of the polysaccharides remained intact following each of these two methods. Compared with AFEX pretreatment, the H-AFEX process enhanced delignification and enzymatic hydrolysis yields of both glucose and xylose. The maximum glucan and xylan digestibility of H-AFEX process were 87.78% and 90.64%, respectively, and were obtained using 0.7 (w/w) water loading, 1.0 (w/w) ammonia loading, 0.5 (w/w) 30wt.% hydrogen peroxide loading, and 130°C for 10min. The results of the present work show that H-AFEX is a feasible pretreatment to improve the enzymatic saccharification of corn stover for bioethanol production.

[Optimization of liquid ammonia treatment for enzymatic hydrolysis of Saccharum arundinaceum to fermentable sugars].

China has abundant available marginal land that can be used for cultivation of lignocellulosic energy plants. Saccharum arundinaceum Retz. is a potential energy crop with both high biomass yield and low soil fertility requirements. It can be planted widely as cellulosic ethanol feedstock in southern China. In the present work Saccharum arundinaceum was pretreated by liquid ammonia treatment (LAT) to overcome biomass recalcitrance, followed by enzymatic hydrolysis. The monosaccharide contents (glucose, xylose, and arabinose) of the enzymatic hydrolysate were determined by high performance liquid chromatography. Experimental results show that the optimal LAT pretreatment conditions were 130 0C, 2:1 (W/W) ammonia to biomass ratio, 80% moisture content (dry weight basis) and 5 min residence time. Approximately 69.34% glucan and 82.60% xylan were converted after 72 h enzymatic hydrolysis at 1% glucan loading using 15 FPU/(g of glucan) of cellulase. The yields of glucose and xylose were 573% and 1 056% higher than those of the untreated biomass, and the LAT-pretreated substrates obtained an 8-fold higher of total monosaccharide yield than untreated substrates. LAT pretreatment was an effective to increase the enzymatic digestibility of Saccharum arundinaceum compared to acid impregnated steam explosion and similar to that of acid treatment and ammonia fiber expansion treatment.

Hydrogen peroxide presoaking of bamboo prior to AFEX pretreatment and impact on enzymatic conversion to fermentable sugars.

Bamboo is a fast growing plant found worldwide that has high potential as an energy crop. This project evaluated the effectiveness of AFEX pretreatment for converting moso bamboo (Phyllostachys heterocycla var. pubescens) to fermentable sugars, both with and without pre-soaking in hydrogen peroxide. Pretreatment conditions including temperature, water loading, residence time, ammonia loading, and hydrogen peroxide loadings were varied to maximize hydrolysis yields. The optimal conditions for AFEX were 150°C, 0.8 or 2.0 (w/w) water loading, 10-30 min residence time, and 2.0-5.0 (w/w) ammonia loading. The optimal conditions for H-AFEX were same AFEX conditions with 0.7-1.9 (w/w) 30% (wt) hydrogen peroxide solutions loading. Using 15 FPU/g glucan cellulase and under optimal conditions, AFEX pretreatment achieved a theoretical sugars yield of 64.8-72.7% and addition of hydrogen peroxide presoaking increased the yield to 83.4-92.1%. It is about 5-fold and 7-fold increase in sugars yield for AFEX-treated and H-AFEX-treated bamboo respectively.

Enzymatic digestibility and ethanol fermentability of AFEX-treated starch-rich lignocellulosics such as corn silage and whole corn plant.

BACKGROUND: Corn grain is an important renewable source for bioethanol production in the USA. Corn ethanol is currently produced by steam liquefaction of starch-rich grains followed by enzymatic saccharification and fermentation. Corn stover (the non-grain parts of the plant) is a potential feedstock to produce cellulosic ethanol in second-generation biorefineries. At present, corn grain is harvested by removing the grain from the living plant while leaving the stover behind on the field. Alternatively, whole corn plants can be harvested to cohydrolyze both starch and cellulose after a suitable thermochemical pretreatment to produce fermentable monomeric sugars. In this study, we used physiologically immature corn silage (CS) and matured whole corn plants (WCP) as feedstocks to produce ethanol using ammonia fiber expansion (AFEX) pretreatment followed by enzymatic hydrolysis (at low enzyme loadings) and cofermentation (for both glucose and xylose) using a cellulase-amylase-based cocktail and a recombinant Saccharomyces cerevisiae 424A (LNH-ST) strain, respectively. The effect on hydrolysis yields of AFEX pretreatment conditions and a starch/cellulose-degrading enzyme addition sequence for both substrates was also studied. RESULTS: AFEX-pretreated starch-rich substrates (for example, corn grain, soluble starch) had a 1.5-3-fold higher enzymatic hydrolysis yield compared with the untreated substrates. Sequential addition of cellulases after hydrolysis of starch within WCP resulted in 15-20% higher hydrolysis yield compared with simultaneous addition of hydrolytic enzymes. AFEX-pretreated CS gave 70% glucan conversion after 72 h of hydrolysis for 6% glucan loading (at 8 mg total enzyme loading per gram glucan). Microbial inoculation of CS before ensilation yielded a 10-15% lower glucose hydrolysis yield for the pretreated substrate, due to loss in starch content. Ethanol fermentation of AFEX-treated (at 6% w/w glucan loading) CS hydrolyzate (resulting in 28 g/L ethanol at 93% metabolic yield) and WCP (resulting in 30 g/L ethanol at 89% metabolic yield) is reported in this work. CONCLUSIONS: The current results indicate the feasibility of co-utilization of whole plants (that is, starchy grains plus cellulosic residues) using an ammonia-based (AFEX) pretreatment to increase bioethanol yield and reduce overall production cost.