The assessment of chromophores in bleached cellulosic pulps employing UV-Raman spectroscopy

UV-Resonance Raman (UV-RR) coupled with UV-visible Diffuse Reflectance (UV-vis DR) spectroscopy was applied to a solid-state study of chromophores in Eucalyptus globulus kraft cellulosic pulps bleached by chlorine dioxide and hydrogen peroxide. The UV-RR spectra were acquired at 325 nm laser beam excitation, which was shown to be appropriate for selective analysis of chromophore structures in polysaccharides. The proposed approach allowed the monitoring of chromophores in pulps and to track the extent of polysaccharide oxidation. However, precaution was suggested while performing a quantitative analysis of chromophores at the characteristic band of ∼1600 cm⁻¹ because of charge transfer complexes (CTCs) that exist in the pulp. These CTCs can affect the intensity of the aforementioned band by diminishing the conjugate state in the chromophore moieties. The amount of carbonyl and carboxyl groups in polysaccharides correlated with the intensity of the band at 1093 cm⁻¹. The analysis of UV-RR spectra revealed xylan as an important source of chromophores in eucalypt kraft pulp.     

Thermophilic anaerobic bacteria which ferment hemicellulose: characterization of organisms and identification of plasmids

Seven thermophilic anaerobic bacteria which ferment xylan were isolated from natural geothermal features in the western United States. Typically, these strains were Gram-negative non-sporeforming rods with an unusual double-layered cell wall which resembled that observed in Thermobacteroides acetoethylicus. The strains differed from known thermophilic anaerobes in their ability to utilize a very wide variety of carbohydrates and in their ability to grow in a chemically-defined medium and/or at pH 3.5. Four of the strains contained cryptic plasmids of 1.2 or 1.5×10⁶ daltons. The taxonomic characteristics of the strains are discussed in terms of their relatedness to those of Thermoanaerobium, Thermoanaerobacter, and Thermobacteroides species.Contribution No. 3222 of the Central Research and Development Department     

A mesophilic Clostridium species that produces butanol from monosaccharides and hydrogen from polysaccharides

A unique mesophilic Clostridium species strain BOH3 is obtained in this study, which is capable of fermenting monosaccharides to produce butanol and hydrolyzing polysaccharides to produce hydrogen (H(2)) and volatile fatty acids (VFAs). From 30 g/L of glucose and xylose each, batch culture BOH3 was able to produce 4.67 and 4.63 g/L of butanol. Enhancement treatments by increasing the inoculated cells improved butanol production to 7.05 and 7.41 g/L, respectively. Hydrogen production (2.47 and 1.93 mmol) was observed when cellulose and xylan (10 g/L each) were used, suggesting that strain BOH3 possesses xylanolytic and cellulolytic capabilities. These unique features reveal the strain's novelty as most wild-type solventogenic strains have not been reported to have such properties. Therefore, culture BOH3 is promising in generating butanol and hydrogen from renewable feedstock.     

Acid-catalyzed conversion of xylose, xylan and straw into furfural by microwave-assisted reaction

Furfural is a biomass derived-chemical that can be used to replace petrochemicals. In this study, the acid-catalyzed conversion of xylose and xylan to furfural by microwave-assisted reaction was investigated at selected ranges of temperature (140-190 °C), time (1-30 min), substrate concentration (1:5-1:200 solid:liquid ratio), and pH (2-0.13). We found that a temperature of 180 °C, a solid:liquid ratio of 1:200, a residence time of 20 min, and a pH of 1.12 gave the best furfural yields. The effect of different Brønsted acids on the conversion efficiency of xylose and xylan was also evaluated, with hydrochloric acid being found to be the most effective catalyst. The microwave-assisted process provides highly efficient conversion: furfural yields obtained from wheat straw, triticale straw, and flax shives were 48.4%, 45.7%, and 72.1%, respectively.     

Study of the intracellular xylanolytic activity of the phytopathogenic fungus Sporisorium reilianum

The present study demonstrates that Sporisorium reilianum, a phytopathogenic fungus of corn, produces intracellular xylanolytic activity during submerged fermentation. Production reached its highest levels in a medium containing glucose, corn hemicellulose and yeast extract. An intracellular xylanase was purified by a process that included precipitation with ammonium sulfate, ion exchange chromatography and gel filtration. Optimal pH and temperature values were 5.0 and 60 °C, respectively. The enzyme showed activity through a broad pH range. The molecular weights of pure xylanase were 36 and 37 kDa, determined by SDS PAGE and gel filtration, respectively. K_m and V_max were 0.160 mg/mL and 1.564 μmol/min/mg, respectively, on a substrate of birchwood xylan. SDS, EDTA, β-Mercaptoethanol, Tween 80, Triton and Mn²⁺ and Ca²⁺ strongly inhibited activity. The purified enzyme hydrolyzed xylan, releasing xylotriose and xylobiose. Sequence protein analysis showed 95% similarity with the theoretical protein encoded by the sr14403 gene of S. reilianum, which encodes a putative endo-β-1,4-xylanase. The enzyme is an isoform of the extracellular xylanase SRXL1 of this basidiomycete.     

A comparative study of antioxidative activities of cell-wall polysaccharides

Oxidative burst in plants is elicited by biotic and abiotic stressors. Analogously to some monosaccharides which act as intracellular antioxidants, cell-wall polysaccharides may be in charge of buffering free-radical production in the extracellular compartment under pronounced prooxidative settings. Although a wide range of plant polysaccharides have been examined for their antioxidative properties, this usually has not been done in a coherent and comparative manner and against biologically relevant reactive species. Here we show that different cell-wall polysaccharides, cellulose, pectin, d-galacto-d-mannan, arabinogalactan, and xylan, exhibit distinctive antioxidative activities against the hydroxyl radical (OH)-generating Fenton reaction and superoxide. We found, using an EPR spin-trapping method, that the main carriers of ‘anti-Fenton’ activity in the plant cell wall are pectin and xylan. They most likely act by binding metal ions in such a manner to allow the Fenton reaction, after which they scavenge OH. Such a mode of action is preferred by cells resulting in a safe degradation of H₂O₂. On the other hand, the polysaccharides examined showed similar superoxide scavenging capacities. We propose that plants may employ different antioxidative characteristics of polysaccharides to regulate their redox status by modifying the composition of the cell wall.     

牦牛源木聚糖酶产生菌的筛选和鉴定

目的以牦牛粪便为样本,筛选并鉴定产木聚糖酶菌株。方法利用碱提取法从玉米芯中提取木聚糖,以自制木聚糖为唯一碳源,从牦牛牛粪中筛选产木聚糖酶细菌,利用16S rDNA基因序列分析鉴定菌种,3,5-二硝基水杨酸法(DNS)测定其产酶能力并分析所产酶的酶学特性。结果筛选获得牦牛源产木聚糖酶类芽胞杆菌,所产木聚糖酶的最适反应条件为50℃、pH 8.0,在pH值为7.0或8.0以及温度50℃条件下,表现出较好的稳定性,Mn~(2+)对酶活力具有显著抑制作用,该菌最佳发酵时间为12 h,酶活最高达到1.2 U/mL。结论该菌所产木聚糖酶能够针对性地降解玉米芯木聚糖,在畜牧业和工业上有一定的应用价值。     

Development of strains of the thermotolerant yeast Hansenula polymorpha capable of alcoholic fermentation of starch and xylan

The thermotolerant yeast Hansenula polymorpha ferments glucose and xylose to ethanol at high temperatures. However, H. polymorpha cannot utilize starchy materials or xylans. Heterologous amylolytic and xylanolytic enzymes have to be expressed in this yeast to provide for utilization and growth on starch and xylan. Genes SWA2 and GAM1 from the yeast Schwanniomyces occidentalis, encoding α-amylase and glucoamylase, respectively, were expressed in H. polymorpha. The expression was achieved by integration of the SWA2 and GAM1 genes under the strong constitutive promoter of the H. polymorpha glyceraldehyde-3-phosphate dehydrogenase gene (HpGAP) into H. polymorpha genome. Resulting transformants acquired the ability to grow on a minimal medium containing soluble starch as a sole carbon source. Ethanol production at high-temperature fermentation from starch by the recombinant strains was up to 10 g/L. The XYN2 gene encoding endoxylanase of the fungus Trichoderma reseei was expressed in H. polymorpha. Co-expression of xlnD gene coding for β-xylosidase of the fungus Aspergillus niger and the XYN2 gene in H. polymorpha was achieved by integration of these genes under control of the HpGAP promoter. Resulting transformants were capable of growth and alcoholic fermentation on a minimal medium supplemented with birchwood xylan as a sole carbon source at 48 °C.     

Quantitative analysis of sugars in wood hydrolyzates with H NMR during the autohydrolysis of hardwoods

The focus of this work was to determine the utility of ¹H NMR spectroscopy in the quantification of sugars resulting from the solubilization of hemicelluloses during the autohydrolysis of hardwoods and the use of this technique to evaluate the kinetics of this process over a range of temperatures and times. Yields of residual xylan, xylooligomers, xylose, glucose, and the degraded products of sugars, i.e., furfural and HMF (5-hydroxymethyl furfural), were determined. The monosaccharide and oligomer contents were quantified with a recently developed high resolution ¹H NMR spectroscopic analysis. This method provided precise measurement of the residual xylan and cellulose remaining in the extracted wood samples and xylose and glucose in the hydrolyzates. NMR was found to exhibit good repeatability and provided carbohydrate compositional results comparable to published methods for sugar maple and aspen woods.     

Water-based woody biorefinery

The conversion of biomass into chemicals and energy is essential in order to sustain our present way of life. Fossil fuels are currently the predominant energy source, but fossil deposits are limited and not renewable. Biomass is a reliable potential source of materials, chemicals and energy that can be replenished to keep pace with our needs. A biorefinery is a concept for the collection of processes used to convert biomass into materials, chemicals and energy. The biorefinery is a “catch and release” method for using carbon that is beneficial to both the environment and the economy. In this study, we discuss three elements of a wood-based biorefinery, as proposed by the SUNY College of Environmental Science and Forestry (ESF): hot-water extraction, hydrolysis, and membrane separation/concentration. Hemicelluloses are the most easily separable main component of woody biomass and thus form the bulk of the extracts obtained by hot-water extraction of woody biomass. Hot-water extraction is an important step in the processes of woody biomass and product generation, replacing alternative costly pre-treatment methods. The hydrolysis of hemicelluloses produces 5-carbon sugars (mainly xylose), 6-carbon sugars (mainly glucose and mannose), and acetic acid. The use of nano-filtration membranes is an efficient technology that can be employed to fractionate hot-water extracts and wood hydrolysate. The residual solid mass after hot-water extraction has a higher energy content and contains fewer easily degradable components. This allows for more efficient subsequent processing to convert cellulose and lignin into conventional products.