Simulation of the ozone pretreatment of wheat straw.

Wheat straw is a potential feedstock in biorefinery for sugar production. However, the cellulose, which is the major source of sugar, is protected by lignin. Ozonolysis deconstructs the lignin and makes cellulose accessible to enzymatic digestion. In this study, the change in lignin concentration with different ozonolysis times (0, 1, 2, 3, 5, 7, 10, 15, 20, 30, 60min) was fit to two different kinetic models: one using the model developed by Garcia-Cubero et al. (2012) and another including an outer mass transfer barrier or "cuticle" region where ozone mass transport is reduced in proportion to the mass of unreacted insoluble lignin in the cuticle. The kinetic parameters of two mathematical models for predicting the soluble and insoluble lignin at different pretreatment time were determined. The results showed that parameters derived from the cuticle-based model provided a better fit to experimental results compared to a model without a cuticle layer.

Structural characterization of lignin: a potential source of antioxidants guaiacol and 4-vinylguaiacol.

The structure of lignin obtained from the ozone and soaking aqueous ammonia pretreatment of wheat straw has been characterized utilizing chemical analytical methods in order to reveal its antioxidant characteristics, including attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS), pyrolysis/tetramethylammonium hydroxide-gas chromatography/mass spectrometry (Py/TMAH-GC/MS), gel permeation chromatography (GPC), ultra violet-visible spectroscopy (UV-vis), and 1,1-diphenyl-2-picrylhydrazyl (DPPH) antioxidant evaluation assay. The results demonstrated that the isolated lignin is a ρ-hydroxyphenyl- guaiacyl-syringyl (H-G-S) lignin, with S/G ratio of 0.35 and significant amounts of phenol 2-methoxy (guaiacol) and phenol 2-methoxy-4-vinyl (4-vinylguaiacol). The Py-GC/MS and Py/TMAH-GC/MS pyrograms indicated that the major units in this lignin are derived from hydroxycinnamic acids. The GPC results revealed the molecular weight of the lignin was considerably low and also the FTIR analysis showed that the lignin possessed hydroxyl and methoxy functional groups; the factors led to the extracted lignin having a comparable antioxidant activity to that of currently used commercial antioxidants. The UV-vis and DPPH antioxidant assay results suggested a percentage of inhibition of the DPPH radicals in the following order: guaiacol (103.6 ± 1.36)>butylated hydroxytoluene (103.3 ± 1)>ferulic acid (102.6 ± 0.79)>pretreated lignin (86.9 ± 0.34).

Enhancing volatile fatty acid (VFA) and bio-methane production from lawn grass with pretreatment.

The bioconversion of fiber-based carbohydrates during anaerobic digestion (AD) is impeded due to the recalcitrant nature of the plant cell wall. Pretreatment of lignocellulose materials under mild conditions are needed to improve the digestibility at minimum cost. This study investigated the effects of different pretreatments, including ozone, soaking aqueous ammonia (SAA), combined ozone and SAA (OSAA), and size reduction to enhance volatile fatty acid (VFA) and bio-methane production when lawn grass was used as substrate. To study VFA production, methanogenesis was selectively inhibited by sodium 2-bromoethanesulfonate to decouple the relation between VFA and bio-methane. The enzymatic hydrolysis of SAA (residence time 24h at 50°C) and OSAA (10 min ozonation and 6h of SAA) in pretreatment of lawn grass sample resulted in 86.71% and 89.63% sugar recovery, respectively. The specific methane yields of the control, ozone, SAA, OSAA, and size-reduced grass samples were 402.5, 358.8, 481.0, 462.6, and 358.3 ml CH4/g volatile solid (VS), respectively.

Structural modification of lignin and characterization of pretreated wheat straw by ozonation.

Ozonolysis is potentially an effective method for pretreating lignocellulosic biomass to improve the production of fermentable sugars via enzymatic hydrolysis. Further understanding of the ozonolysis process and identifying specific lignin structural changes are crucial for improving the pretreatment process. Investigation into pretreatment of wheat straw using ozonolysisis is reported in this paper, with special emphasis on selective modification/degradation of lignin subunits. The ozonolysis was performed for 2 h with less than 60 mesh particles in order to achieve maximum lignin oxidation. The results showed that the lignin structure was significantly modified under these conditions, leading to higher sugar recovery of more than 50% which increased from 13.11% to 63.17% corresponding to the control and ozone treated samples, respectively. Moisture content was found to be an important parameter for improving sugar recovery. Ninety percent (w/w) moisture produced the highest sugar recovery. The concentration of acid soluble lignin in the ozone treated sample increased from 4% to 11% after 2 h treatment. NMR analysis revealed that the S2/6 and G2 lignin units in the wheat straw were most prone to oxidation by ozone as the concentration of aromatic units decreased while the carboxylic acids became more abundant. The experimental data suggest the degradation of ß-O-4 moieties and aromatic ring opening in lignin subunits. The pyrolysis-gas chromatography/mass spectrometry results revealed that the rate of lignin unit degradation was in the following order: syringyl > guaiacyl > p-hydroxyphenyl. Long ozone exposure resulted in few condensed lignin structure formation. In addition, the formation of condensed units during this process increased the activation energy from ASTM-E, 259.74 kJ/mol; Friedman-E, 270.08 kJ/mol to ASTM-E, 509.29 kJ/mol; Friedman-E, 462.17 kJ/mol. The results provide new information in overcoming lignin barrier for lignocellulose utilization.

Structural and thermal characterization of wheat straw pretreated with aqueous ammonia soaking.

Production of renewable fuels and chemicals from lignocellulosic feedstocks requires an efficient pretreatment technology to allow ready access of polysaccharides for cellulolytic enzymes during saccharification. The effect of pretreatment on wheat straw through a low-temperature and low-pressure soaking aqueous ammonia (SAA) process was investigated in this study using Fourier transform infrared (FTIR), pyrolysis-gas chromatography/mass spectroscopy (Py-GC/MS), solid and liquid state nuclear magnetic resonance (NMR), and thermogravimetry/differential thermogravimetry (TG/DTG) to demonstrate the changes in lignin, hemicellulose, and cellulose structure. After treatment of 60 mesh wheat straw particles for 60 h with 28-30% ammonium hydroxide (1:10 solid/liquid) at 50 °C, sugar recovery increased from 14% (untreated) to 67% (SAA treated). The FTIR study revealed a substantial decrease in absorbance of lignin peaks. Solid and liquid state NMR showed minimal lignin structural changes with significant compositional changes. Activation energy of control and pretreated wheat straw was calculated according to the Friedman and ASTM methods and found to be decreased for SAA-treated wheat straw, from 259 to 223 kJ/mol. The SAA treatment was shown to remove significant amounts of lignin without strongly affecting lignin functional groups or structure.

Development of a protocol for supercritical carbon dioxide extraction of ubiquinone-10 from dried biomass of Pseudomonas diminuta.

Ubiquinone (Coenzyme Q10, CoQ10), a yellow-to-orange-colored lipophilic substance having nutraceutical value, was extracted from dried biomass of Pseudomonas diminuta using supercritical carbon dioxide (SC-CO(2)). The effect of different operational parameters (temperature, pressure, and extraction time) and addition of co-solvent on SC-CO(2) extraction of CoQ10 was studied in detail. The solubility parameter of CoQ10, CO(2), and CO(2) with ethanol and methanol as co-solvents was calculated and validated with experimental results. Theoretically, ethanol and methanol had significant effect as co-solvent, and the difference between the two was only marginal. A maximum recovery of 22.33% was obtained using pure SC-CO(2) at 40 °C, 150 bar, and run time of 60 min. Ethanol as co-solvent at 3 mL/g of dried biomass at 40 °C and 150 bar increased the recovery from 22.33 to 68.57%. Further optimization of the extraction conditions by Box-Behnken design effectively increased the recovery to 96.2%. The optimized conditions were a temperature of 38 °C, pressure of 215 bar, and run time of 58 min.

Production of biovanillin by one-step biotransformation using fungus Pycnoporous cinnabarinus.

The current study proposes a one-step biotransformation process for vanillin production from ferulic acid using the wild fungal strain Pycnoporous cinnabarinus belonging to the family Basidiomycete. Improvement of biotransformation conditions was performed in two steps; initially a one factor at a time method was used to investigate effects of medium composition variables (i.e., carbon, nitrogen) and environmental factors such as pH on vanillin production. Subsequently, concentrations of medium components were optimized using an orthogonal matrix method. After primary screening, glucose as carbon source and corn steep liquor and ammonium chloride as organic and inorganic nitrogen source, respectively, supported maximum biotransformation of ferulic acid to vanillin. Under statistically optimum conditions vanillin production from ferulic acid by P. cinnabarinus was 126 mg/L with a molar yield of 54%. The overall molar yield of vanillin production increased by 4 times.

Glucose oxidase--an overview.

Glucose oxidase (beta-D-glucose:oxygen 1-oxidoreductase; EC 1.1.2.3.4) catalyzes the oxidation of beta-D-glucose to gluconic acid, by utilizing molecular oxygen as an electron acceptor with simultaneous production of hydrogen peroxide. Microbial glucose oxidase is currently receiving much attention due to its wide applications in chemical, pharmaceutical, food, beverage, clinical chemistry, biotechnology and other industries. Novel applications of glucose oxidase in biosensors have increased the demand in recent years. Present review discusses the production, recovery, characterization, immobilization and applications of glucose oxidase. Production of glucose oxidase by fermentation is detailed, along with recombinant methods. Various purification techniques for higher recovery of glucose oxidase are described here. Issues of enzyme kinetics, stability studies and characterization are addressed. Immobilized preparations of glucose oxidase are also discussed. Applications of glucose oxidase in various industries and as analytical enzymes are having an increasing impact on bioprocessing.

Preparation of ferulic acid from agricultural wastes: its improved extraction and purification.

Ferulic acid (FA) is a phenolic antioxidant present in plants, which is widely used in the food and cosmetic industry. In the present study, various agricultural wastes such as maize bran, rice bran, wheat bran, wheat straw, sugar cane baggasse, pineapple peels, orange peels, and pomegranate peels were screened for the presence of esterified FA (EFA). Among the sources screened, maize bran was found to contain the highest amount of EFA. Pineapple peels, orange peels, and pomegranate peels were also found to contain traces of EFA. Alkaline extraction of EFA from maize bran was carried out using 2 M NaOH. Response surface methodology (RSM) was used for optimization of EFA extraction, which resulted in a 1.3-fold increase as compared to the unoptimized conventional extraction technique. FA was analyzed by means of high-performance liquid chromatography (HPLC). Purification was carried out by adsorption chromatography using Amberlite XAD-16 followed by preparative high-performance thin-layer chromatography (HPTLC). The recovery of Amberlite XAD-16 purified FA was up to 57.97% with HPLC purity 50.89%. The fold purity achieved was 1.35. After preparative HPTLC, the maximum HPLC purity obtained was 95.35% along with an increase in fold purity up to 2.53.