Sulfonated hierarchical H-USY zeolite for efficient hydrolysis of hemicellulose/cellulose

Sulfonated hierarchical H-USY zeolite was prepared and characterized by X-ray diffraction, N₂ physisorption, Fourier transform infrared spectroscopy, inductively coupled plasma atomic emission spectroscopy, temperature-programmed desorption of ammonia, and acid–base titration. It was proved that sulfonic group was successfully anchored onto the hierarchical H-USY zeolite. The acidity of the hierarchical H-USY was remarkably improved. Sulfonated hierarchical H-USY zeolite was efficient for the hydrolysis of hemicellulose and cellulose. The yield of TRS for hydrolysis of hemicellulose reached 78.0% at 140 °C for 9 h. For hydrolysis of α-cellulose, 60.8% conversion with 22.4% yield of glucose was obtained. Even for microcrystalline cellulose, 43.7% conversion with 15.1% yield of glucose can be obtained. These results are much higher than those obtained over hierarchical H-USY zeolite, indicating that both the acidity and the pore structure determine the activity of zeolite as catalyst in the hydrolysis of biomass.     

Xylanase and feruloyl esterase from actinomycetes cultures could enhance sugarcane bagasse hydrolysis in the production of fermentable sugars

The addition of enzymes that are capable of degrading hemicellulose has a potential to reduce the need for commercial enzymes during biomass hydrolysis in the production of fermentable sugars. In this study, a high xylanase producing actinomycete strain (Kitasatospora sp. ID06-480) and the first ethyl ferulate producing actinomycete strain (Nonomuraea sp. ID06-094) were selected from 797 rare actinomycetes, respectively, which were isolated in Indonesia. The addition (30%, v/v) of a crude enzyme supernatant from the selected strains in sugarcane bagasse hydrolysis with low-level loading (1 FPU/g-biomass) of Cellic® CTec2 enhanced both the released amount of glucose and reducing sugars. When the reaction with Ctec2 was combined with crude enzymes containing either xylanase or feruloyl esterase, high conversion yield of glucose from cellulose at 60.5% could be achieved after 72 h-saccharification.     

Development and validation of a kinetic model for enzymatic saccharification of lignocellulosic biomass

A multireaction kinetic model was developed for closed-system enzymatic hydrolysis of lignocellulosic biomass such as corn stover. Three hydrolysis reactions were modeled, two heterogeneous reactions for cellulose breakdown to cellobiose and glucose and one homogeneous reaction for hydrolyzing cellobiose to glucose. Cellulase adsorption onto pretreated lignocellulose was modeled via a Langmuir-type isotherm. The sugar products of cellulose hydrolysis, cellobiose and glucose, as well as xylose, the dominant sugar prevalent in most hemicellulose hydrolyzates, were assumed to competitively inhibit the enzymatic hydrolysis reactions. Model parameters were estimated from experimental data generated using dilute acid pretreated corn stover as the substrate. The model performed well in predicting cellulose hydrolysis trends at experimental conditions both inside and outside the design space used for parameter estimation and can be used for in silico process optimization.     

Fermentability of the hemicellulose‐derived sugars from steam‐exploded softwood (douglas fir)

Steam explosion ofDouglas fir wood chips under low‐severity conditions (log Ro = 3.08 corresponding to 175°C, 7.5 min, and 4.5% SO₂) resulted in the recovery of around 87% of the original hemicellulose component in the water‐soluble stream. More than 80% of the recovered hemicellulose was in a monomeric form. As the pretreatment severity increased from 3.08 to 3.76, hemicellulose recovery dropped to 43% of the original hemicellulose found in Douglas fir chips while the concentration of glucose originating from cellulose hydrolysis increased along with the concentration of sugar degradation products such as furfural and hydroxymethylfurfural. Despite containing a higher concentration of hexose monomers (mainly glucose originating from cellulose degradation), the water‐soluble fraction prepared under high‐severity conditions (log Ro = 3.73 corresponding to 215°C, 2.38 min, and 2.38% SO₂) was not readily fermented. Only the two hydrolyzates obtained at low and medium (195°C, 4.5 min, and 4.5% SO₂) severities were fermented to ethanol using a spent sulfur liquor adapted strain of Saccharomyces cerevisiae. High ethanol yields were obtained for these two hydrolyzates with 0.44 g of ethanol produced per gram of hexose utilized (86% of theoretical). However, the best results of hemicellulose recovery and fermentability were obtained for the low‐severity water‐soluble fraction which was fermented significantly faster than the fraction obtained after medium‐severity treatment probably because it contained higher amounts of fermentation inhibitors. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 64: 284–289, 1999.     

Detection of Metabolic Key Enzymes of Methane Turnover Processes in Cold Seep Microbial Biofilms

In anoxic environments, methane oxidation is conducted in a syntrophic process between methanotrophic archaea (ANME) and sulfate reducing bacteria (SRB). Microbial mats consisting of ANME, SRB and other microorganisms form methane seep-related carbonate buildups in the anoxic bottom waters of the Black Sea Crimean shelf. To shed light on the localization of the biochemical processes at the level of single cells in the Black Sea microbial mats, we applied antibody-based markers for key enzymes of the relevant metabolic pathways. The dissimilatory adenosine-5′-phosphosulfate (APS) reductase, methyl-coenzyme M reductase (MCR) and methanol dehydrogenase (MDH) were selected to localize sulfate respiration, reverse methanogenesis and aerobic methane oxidation, respectively. The key enzymes could be localized by double immunofluorescence and immunocytochemistry at light- and electron microscopic levels. In this study we show that sulfate reduction is conducted synchronized and in direct proximity to reverse methanogenesis of ANME archaea. Microcolonies in interspaces between ANME/SRB express methanol dehydrogenase, which is indicative for oxidation of C₁ compounds by methylotrophic or methanotrophic bacteria. Thus, in addition to syntrophic AOM, oxygen-dependent processes are also conducted by a small proportion of the microbial population.     

Anaerobic biodegradability of cellulose and hemicellulose in excavated refuse samples using a biochemical methane potential assay

Summary Improved techniques are needed to predict potential methane generation from refuse buried in landfills. The Biochemical Methane Potential (BMP) test was used to measure the methane potential of ten refuse samples excavated from a Berkeley, CA, landfill. The test was conducted in 125-ml serum bottles containing phosphate-buffered medium and inoculated with anaerobically digested sewage sludge. Comparison of the measured BMP to the theoretical BMP calculated from measured cellulose and hemicellulose concentrations indicated that cellulose plus hemicellulose is not well correlated with the measured BMP. The average of the measured to theoretical BMP was 19.1% (range 0–53%, s.d.=16.9%). Measured sulfate concentrations showed that sulfate was an insignificant electron sink in the samples tested. Once methane production from the refuse was complete, 0.072 g of Whatman no. 1 filter paper was added to two of the four serum bottles incubated for each sample. An average of 84.9% (s.d=2.5%) of the added filter paper was recovered as methane, suggesting that some cellulose and hemicellulose present in refuse is recalcitrant or otherwise not bioavailable.     

Metabolic associations with archaea drive shifts in hydrogen isotope fractionation in sulfate‐reducing bacterial lipids in cocultures and methane seeps

Correlation between hydrogen isotope fractionation in fatty acids and carbon metabolism in pure cultures of bacteria indicates the potential of biomarker D/H analysis as a tool for diagnosing carbon substrate usage in environmental samples. However, most environments, in particular anaerobic habitats, are built from metabolic networks of micro‐organisms rather than a single organism. The effect of these networks on D/H of lipids has not been explored and may complicate the interpretation of these analyses. Syntrophy represents an extreme example of metabolic interdependence. Here, we analyzed the effect of metabolic interactions on the D/H biosignatures of sulfate‐reducing bacteria (SRB) using both laboratory maintained cocultures of the methanogen Methanosarcina acetivorans and the SRB Desulfococcus multivorans in addition to environmental samples harboring uncultured syntrophic consortia of anaerobic methane‐oxidizing archaea (ANME) and sulfate‐reducing Deltaproteobacteria (SRB) recovered from deep‐sea methane seeps. Consistent with previously reported trends, we observed a ~80‰ range in hydrogen isotope fractionation (ε_{lipid–water}) for D. multivorans grown under different carbon assimilation conditions, with more D‐enriched values associated with heterotrophic growth. In contrast, for cocultures of D. multivorans with M. acetivorans, we observed a reduced range of ε_{lipid–water} values (~36‰) across substrates with shifts of up to 61‰ compared to monocultures. Sediment cores from methane seep settings in Hydrate Ridge (offshore Oregon, USA) showed similar D‐enrichment in diagnostic SRB fatty acids coinciding with peaks in ANME/SRB consortia concentration suggesting that metabolic associations are connected to the observed shifts in ε_{lipid–water} values.     

Major substrates for microbial sulfate reduction in the sediments of Ise Bay, Japan

To clarify the anaerobic microbial interactions in the process of carbon mineralization in marine eutrophic environments, the microbial sulfate reduction and methane production rates were examined in coastal marine sediments of Ise Bay, Japan, in autumn 1990. Sulfate reduction rates (51–210 nmol ml⁻¹ day⁻¹ at 24°C) were much higher than the methane production ones (<1.78 nmol ml⁻¹ day⁻¹) in the surface sediments (top 2 cm) at the six stations surveyed (water depth: 10.7–23.3 m). Substrates for sulfate-reducing bacteria (SRB) were estimated after the addition of a specific inhibitor for SRB (20 mmol l⁻¹ molybdate) into the sediment slurry, from the substrate accumulation rates. In the presence of the inhibitor, sulfate reduction was completely stopped and volatile fatty acids (mainly acetate) were accumulated, although hydrogen was not. Methane production occurred markedly accompanied by consumption of the accumulated acetate from the third day after the addition of molybdate. The maximum rate of methane production was 1.2–1.9 μmol ml⁻¹ day⁻¹, which was similar to those in highly polluted freshwater sediments such as the Tama River, Tokyo, Japan. These results show that acetate is a common major substrate for sulfate reduction and methane production, and SRB competitively inhibit potential acetoclastic methanogenesis in coastal sediments. Methanogens may potentially inhabit the sediments at low levels of population density and activity.     

Solid state fermentation of oat straw by Polyporus sp A-336 and the effect of added sugars

Summary Supplementing oat straw in SSF by Polyporus sp A-336 with xylose, mannose, glucose and arabinogalactan at levels of 5 and 10% of straw weight stimulated lignin degradation and cellulose hydrolysis. Degradation of lignin, hemicellulose and cellulose was monitored for 30 days in plain straw, and straw plus xylose and showed that xylose shortened the lag in lignin breakdown and slowed hemicellulose utilization. At 24 days, similar polymer losses occurred in both systems and enzymatic cellulose hydrolysis had reached a maximum of 47% weight loss.     

Integrated metagenomic and metaproteomic analyses of an ANME-1-dominated community in marine cold seep sediments

Sulfate‐reducing methanotrophy by anaerobic methanotrophic archaea (ANME) and sulfate‐reducing bacteria (SRB) is a major biological sink of methane in anoxic methane‐enriched marine sediments. The physiology of a microbial community dominated by free‐living ANME‐1 at 14–16 cm below the seafloor in the G11 pockmark at Nyegga was investigated by integrated metagenomic and metaproteomic approaches. Total DNA was subjected to 454‐pyrosequencing (829 527 reads), and 16.6 Mbp of sequence information was assembled into 27352 contigs. Taxonomic analysis supported a high abundance of Euryarchaea (70%) with 66% of the assembled metagenome belonging to ANME‐1. Extracted sediment proteins were separated in two dimensions and subjected to mass spectrometry (LTQ‐Orbitrap XL). Of 356 identified proteins, 245 were expressed by ANME‐1. These included proteins for cold‐adaptation and production of gas vesicles, reflecting both the adaptation of the ANME‐1 community to a permanently cold environment and its potential for positioning in specific sediment depths respectively. In addition, key metabolic enzymes including the enzymes in the reverse methanogenesis pathway (except N⁵,N¹⁰‐methylene‐tetrahydromethanopterin reductase), heterodisulfide reductases and the F₄₂₀H₂:quinone oxidoreductase (Fqo) complex were identified. A complete dissimilatory sulfate reduction pathway was expressed by sulfate‐reducing Deltaproteobacteria. Interestingly, an APS‐reductase comprising Gram‐positive SRB and related sequences were identified in the proteome. Overall, the results demonstrated that our approach was effective in assessing in situ metabolic processes in cold seep sediments.     

Pretreatment of wheat straw using combined wet oxidation and alkaline hydrolysis resulting in convertible cellulose and hemicellulose

The wet oxidation process of wheat straw has been studied as a pretreatment method to attain our main goal: To break down cellulose to glucose enzymatic, and secondly, to dissolve hemicellulose (e.g., for fermentation) without producing microbial inhibitors. Wet oxidation combined with base addition readily oxidizes lignin from wheat straw facilitating the polysaccharides for enzymatic hydrolysis. By using a specially constructed autoclave system, the wet oxidation process was optimized with respect to both reaction time and temperature. The best conditions (20 g/L straw, 170 degrees C, 5 to 10 min) gave about 85% w/w yield of converting cellulose to glucose. The process water, containing dissolved hemicellulose and carboxylic acids, has proven to be a direct nutrient source for the fungus Aspergillus niger producing exo-beta-xylosidase. Furfural and hydroxymethyl-furfural, known inhibitors of microbial growth when other pretreatment systems have been applied, were not observed following the wet oxidation treatment. (c) 1996 John Wiley & Sons, Inc.     

Catalytic conversion of cellulose to chemicals in ionic liquid

A simple and effective route for the production of 5-hydroxymethyl furfural (HMF) and furfural from microcrystalline cellulose (MCC) has been developed. CoSO₄ in an ionic liquid, 1-(4-sulfonic acid) butyl-3-methylimidazolium hydrogen sulfate (IL-1), was found to be an efficient catalyst for the hydrolysis of cellulose at 150 °C, which led to 84% conversion of MCC after 300 min reaction time. In the presence of a catalytic amount of CoSO₄, the yields of HMF and furfural were up to 24% and 17%, respectively; a small amount of levulinic acid (LA) and reducing sugars (8% and 4%, respectively) were also generated. Dimers of furan compounds were detected as the main by-products through HPLC-MS, and with the help of mass spectrometric analysis, the components of gas products were methane, ethane, CO, CO_2, and H₂. A mechanism for the CoSO₄-IL-1 hydrolysis system was proposed and IL-1 was recycled for the first time, which exhibited favorable catalytic activity over five repeated runs. This catalytic system may be valuable to facilitate energy-efficient and cost-effective conversion of biomass into biofuels and platform chemicals.     

Kinetic studies of corn stover saccharification using sulphuric acid

The kinetics of crystalline cellulose and hemicellulose hydrolysis in corn stover were studied with a nonisothermal technique. Reactions were arrested at temperatures between 160 and 240 degrees C and product sugars were analyzed using a Bio-Rad HPX-85 liquid chromatographic column. A simple first-order series reaction model was used for both cellulose and hemicellulose hydrolysis reactions. Kinetic parameters were obtained for three different sulphuric acid concentrations (0.49, 0.92, and 1.47 wt %). Activation energies remained constant over this acid concentration range but the preexponential factors showed an increase with acid concentration. Relationships were obtained between the preexponential factors and acid concentrations. Cellulose hydrolysis and glucose degradation reactions were observed to be of higher order with respect to acid concentration in comparison with the previous studies with other raw materials.     

Influence of xylan on the enzymatic hydrolysis of steam‐pretreated corn stover and hybrid poplar

The focus of this study was to alter the xylan content of corn stover and poplar using SO₂‐catalyzed steam pretreatment to determine the effect on subsequent hydrolysis by commercial cellulase preparations supplemented with or without xylanases. Steam pretreated solids with xylan contents ranging from ～1 to 19% (w/w) were produced. Higher xylan contents and improved hemicellulose recoveries were obtained with solids pretreated at lower severities or without SO₂‐addition prior to pretreatment. The pretreated solids with low xylan content (<4% (w/w)) were characterized by fast and complete cellulose to glucose conversion when utilizing cellulases. Commercial cellulases required xylanase supplementation for effective hydrolysis of pretreated substrates containing higher amounts of xylan. It was apparent that the xylan content influenced both the enzyme requirements for hydrolysis and the recovery of sugars during the pretreatment process. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009     

Hydrolysis of lignocellulosic materials for ethanol production: a review

Lignocellulosic biomass can be utilized to produce ethanol, a promising alternative energy source for the limited crude oil. There are mainly two processes involved in the conversion: hydrolysis of cellulose in the lignocellulosic biomass to produce reducing sugars, and fermentation of the sugars to ethanol. The cost of ethanol production from lignocellulosic materials is relatively high based on current technologies, and the main challenges are the low yield and high cost of the hydrolysis process. Considerable research efforts have been made to improve the hydrolysis of lignocellulosic materials. Pretreatment of lignocellulosic materials to remove lignin and hemicellulose can significantly enhance the hydrolysis of cellulose. Optimization of the cellulase enzymes and the enzyme loading can also improve the hydrolysis. Simultaneous saccharification and fermentation effectively removes glucose, which is an inhibitor to cellulase activity, thus increasing the yield and rate of cellulose hydrolysis.     

微生物烟气脱硫中硫酸盐还原阶段的限制性因素及其影响

【目的】利用硫酸盐还原菌(SRB)厌氧活性污泥进行烟气脱硫,探索硫酸盐生物还原的最适条件及重金属离子对硫酸盐生物还原的影响,以提高硫酸盐还原阶段的效率。【方法】对取自污水处理厂的SRB厌氧活性污泥进行高浓度硫酸盐胁迫驯化。分析生物脱硫过程中SRB厌氧污泥还原硫酸盐的限制性因素及影响。【结果】在最适生长条件下(pH 6.5,32°C),经驯化获得的SRB厌氧活性污泥有较强的硫酸盐还原能力。Fe2+的适量添加对硫酸盐还原有一定促进作用。SRB厌氧污泥还原硫酸盐的ThCOD/SO42-最适值为3.00,ThCOD=3.33为最适理论化学需氧量,硫酸盐还原率可达72.15%。SRB厌氧污泥还原硫酸盐反应体系中抑制SRB活性的硫化物浓度为300 mg/L。Pb2+和Ni2+在较低的浓度下(1.0 mg/L和2.0 mg/L)对硫酸盐的还原产生较强的抑制作用,而Cu2+在稍高的浓度下(8.0 mg/L)显示出明显的抑制作用。【结论】经驯化,SRB厌氧活性污泥显示出较强的硫酸盐还原能力,具有应用于工业烟气生物脱硫的潜力。去除重金属离子Pb2+、Ni2+和Cu2+可有效解除对硫酸盐生物还原作用的抑制。     

Methylmercury and methane production potentials in North Carolina Piedmont stream sediments

Methylated mercury (MeHg) can be produced by all microbes possessing the genes hgcA and hgcB, which can include sulfate-reducing bacteria (SRB), iron-reducing bacteria (FeRB), methane-producing archaea (MPA), and other anaerobic microbes. These microbial groups compete for substrates, including hydrogen and acetate. When sulfate is in excess, SRB can outcompete other anaerobic microbes. However, low concentrations of sulfate, which often occur in stream sediments, are thought to reduce the relative importance of SRB. Although SRB are regarded as the primary contributors of MeHg in many aquatic environments, their significance may not be universal, and stream sediments are poorly studied with respect to microbial Hg methylation. We evaluated suppression of methanogenesis by SRB and the potential contributions from SRB, MPA and other MeHg producing microbes (including FeRB) to the production of MeHg in stream sediments from the North Carolina Piedmont region. Lower methanogenesis rates were observed when SRB were not inhibited, however, application of a sulfate-reduction inhibitor stimulated methanogenesis. Greater MeHg production occurred when SRB were active. Other MeHg producing microbes (i.e., FeRB) contributed significantly less MeHg production than SRB. MPA produced MeHg in negligible amounts. Our results suggest that SRB are responsible for the majority of MeHg production and suppress methanogenesis in mid-order stream sediments, similar to other freshwater sediments. Further investigation is needed to evaluate the generality of these findings to streams in other regions, and to determine the mechanisms regulating sulfate and electron acceptor availability and other potential factors governing Hg methylation and methane production in stream sediments.     

Bioethanol,biohydrogen and biogas production from wheat straw in a biorefinery concept

The production of bioethanol, biohydrogen and biogas from wheat straw was investigated within a biorefinery framework. Initially, wheat straw was hydrothermally liberated to a cellulose rich fiber fraction and a hemicellulose rich liquid fraction (hydrolysate). Enzymatic hydrolysis and subsequent fermentation of cellulose yielded 0.41 g-ethanol/g-glucose, while dark fermentation of hydrolysate produced 178.0 ml-H₂/g-sugars. The effluents from both bioethanol and biohydrogen processes were further used to produce methane with the yields of 0.324 and 0.381 m³/kg volatile solids (VS)_added, respectively. Additionally, evaluation of six different wheat straw-to-biofuel production scenaria showed that either use of wheat straw for biogas production or multi-fuel production were the energetically most efficient processes compared to production of mono-fuel such as bioethanol when fermenting C6 sugars alone. Thus, multiple biofuels production from wheat straw can increase the efficiency for material and energy and can presumably be more economical process for biomass utilization.     

Isolation and characterization of a furfural degrading sulfate-reducing bacterium from an anaerobic digester

A sulfate-reducing bacterium (SRB) was isolated from a continuous anaerobic digester, which converted the furfural-containing wastewater to methane and CO₂. This SRB isolate could use furfural, furfuryl alcohol, and 2-furoic acid as sole source of carbon and energy in a defined mineral sulfate medium. Acetic acid was the major end product of furfural degradation. This organism also used wide varieties of other carbon sources, including ethanol, pyruvate, lactate, succinate, propanol, formate, and malate. The SRB isolate contained the electron carrier desulfoviridin. It used SO₄, NO₃, and thiosulfate as electron acceptors. This isolate used ammonium chloride, nitrate and glutamate as nitrogen source. The characteristics of the SRB isolate were closely similar toDesulfovibrio sp.