Extrusion of xylans extracted from corn cobs into biodegradable polymeric materials.

Solvent casting technique, which comprises multiple energy demanding steps including the dissolution of a polymer in a solvent followed by the evaporation of the solvent from the polymer solution, is currently the main technique for the production of xylan based polymeric materials. The present study shows that sufficient water content renders arabinoglucuronoxylan (AGX) polymers extrudable, enabling the production of AGX based polymeric materials in a single step via extrusion, which is economically advantageous to solvent casting process for mass production. AGX polymers with water content of 27% were found to yield extrudates at an extrusion temperature of 90°C. The extruded strips showed very good mechanical properties with an ultimate tensile strength of 76 ± 6 MPa and elongation at break value of 35 ± 8%, which were superior to the mechanical properties of the strips obtained from polylactic acid.

Structure, recombinant expression and mutagenesis studies of the catalase with oxidase activity from Scytalidium thermophilum.

Scytalidium thermophilum produces a catalase with phenol oxidase activity (CATPO) that catalyses the decomposition of hydrogen peroxide into oxygen and water and also oxidizes various phenolic compounds. A codon-optimized catpo gene was cloned and expressed in Escherichia coli. The crystal structures of native and recombinant S. thermophilum CATPO and two variants, H82N and V123F, were determined at resolutions of 2.7, 1.4, 1.5 and 1.9 Å, respectively. The structure of CATPO reveals a homotetramer with 698 residues per subunit and with strong structural similarity to Penicillium vitale catalase. The haem component is cis-hydroxychlorin γ-spirolactone, which is rotated 180° with respect to small-subunit catalases. The haem-binding pocket contains two highly conserved water molecules on the distal side. The H82N mutation resulted in conversion of the native d-type haem to a b-type haem. Kinetic studies of the H82N and V123F mutants indicate that both activities are likely to be associated with the haem centre and suggest that the secondary oxidase activity may be a general feature of catalases in the absence of hydrogen peroxide.

Enhancing the value of nitrogen from rapeseed meal for microbial oil production.

Rapeseed meal, a major byproduct of biodiesel production, has been used as a low-cost raw material for the production of a generic microbial feedstock through a consolidated bioconversion process. Various strategies were tested for the production of a novel fermentation medium, rich in free amino nitrogen (FAN): commercial enzymes (CEs) (2.7 mg g⁻¹ dry meal), liquid state fungal pre-treatment (LSF) using Aspergillus oryzae (4.6 mg g⁻¹), liquid state fungal pre-treatment followed by fungal autolysis (LSFA) (9.13 mg g⁻¹), liquid state pre-treatment using fungal enzymatic broth (EB) (2.1 mg g⁻¹), but the best strategy was a solid state fungal pre-treatment followed by fungal autolysis (34.5 mg g⁻¹). The bioavailability of the nitrogen sources in the novel medium was confirmed in fed-batch bioreactor studies, in which 82.3g dry cell L⁻¹ of the oleaginous yeast Rhodosporidium toruloides Y4 was obtained with a lipid content of 48%. The dry cell weight obtained was higher than that obtained using conventional yeast extract, due to a higher total nitrogen content in the novel biomedium. The fatty acids obtained from the microbial oil were similar to those derived from rapeseed oil.

Evaluation of alkaline pretreatment temperature on a multi-product basis for the co-production of glucose and hemicellulose based films from lignocellulosic biomass.

Cotton stalks were subjected to alkaline pretreatment for the co-production of glucose and hemicellulose based films with a multi-product approach. Three pretreatment temperatures (25, 60 and 90 °C) were evaluated for their effects both on the glucose yield and on the properties of hemicellulose based films. Compared to untreated cotton stalks, the glucose yields were enhanced 3.9, 4.1 and 4.2 times for pretreatments conducted at 25, 60 and 90 °C, respectively. The pretreatment temperature of 90 °C was detrimental in terms of film formation. Tensile energy to break values of the films obtained after pretreatments conducted at 25, 60 and 90 °C were 1.1, 0.8, and 0.4 MJ/m3, respectively. The hemicellulosic part of the process, which considers the production of hemicellulose based films, should govern the pretreatment temperature since it was more responsive to the changes in the pretreatment temperature compared to the cellulosic part that accounts for glucose production.

Selecting the right blood glucose monitor for the determination of glucose during the enzymatic hydrolysis of corncob pretreated with different methods.

In order to assess their accuracy for the determination of glucose during the enzymatic hydrolysis of pretreated lignocellulosic biomass, four different blood glucose monitors (BGMs), each utilizing a different enzymatic mechanism for the determination of glucose, were utilized in an experimental setup, which compares the efficiency of ionic liquid pretreatment with dilute acid and alkaline pretreatments applied on corncob. Among the tested devices, Optium Xceed was found to be the most accurate device for the determination of glucose where Accu-Chek Active was the least accurate BGM, yielding similar results to those obtained with DNS method. Based on the HPLC results, the % error values for Optium Xceed ranged between 3.9-10.5% for the determination of glucose concentration. Upon enzymatic hydrolysis, ionic liquid and alkaline pretreatments gave similar glucose yields, which were slightly higher than the dilute acid pretreatment, which were 31.9%, 31.0% and 27.8%, respectively, based on untreated corncob.

The optimization of dilute acid hydrolysis of cotton stalk in xylose production.

Cotton stalk, a lignocellulosic waste material, is composed of xylose that can be used as a raw material for production of xylitol, a high-value product. There is a growing interest in the use of lignocellulosic wastes for conversion into various chemicals because of their low cost and the fact that they are renewable and abundant. The objective of the study was to determine the effects of H(2)SO(4) concentration, temperature, and reaction time on the production of sugars (xylose, glucose, and arabinose) and on the reaction by-products (furfural and acetic acid). Response surface methodology was used to optimize the hydrolysis process in order to obtain high xylose yield and selectivity. The optimum reaction temperature, reaction time, and acid concentration were 140 °C, 15 min, and 6%, respectively. Under these conditions, xylose yield and selectivity were found to be 47.88% and 2.26 g g(-1), respectively.

Crystallization and preliminary X-ray analysis of a bifunctional catalase-phenol oxidase from Scytalidium thermophilum.

Catalase-phenol oxidase from Scytalidium thermophilum is a bifunctional enzyme: its major activity is the catalase-mediated decomposition of hydrogen peroxide, but it also catalyzes phenol oxidation. To understand the structural basis of this dual functionality, the enzyme, which has been shown to be a tetramer in solution, has been purified by anion-exchange and gel-filtration chromatography and has been crystallized using the hanging-drop vapour-diffusion technique. Streak-seeding was used to obtain larger crystals suitable for X-ray analysis. Diffraction data were collected to 2.8 A resolution at the Daresbury Synchrotron Radiation Source. The crystals belonged to space group P2(1) and contained one tetramer per asymmetric unit.

Cloning, expression and characterization of endo-beta-1,4-mannanase from Aspergillus fumigatus in Aspergillus sojae and Pichia pastoris.

To be utilized in biomass conversion, including ethanol production and galactosylated oligosaccharide synthesis, namely prebiotics, the gene of extracellular endo-beta-1,4-mannanase (EC 3.2.1.78) of Aspergillus fumigatus IMI 385708 (formerly known as Thermomyces lanuginosus IMI 158749) was expressed first in Aspergillus sojae and then in Pichia pastoris under the control of the glyceraldehyde triphosphate dehydrogenase (gpdA) and the alcohol oxidase (AOX1) promoters, respectively. The highest production of mannanase (352 U mL(-1)) in A. sojae was observed after 6 days of cultivation. In P. pastoris, the highest mannanase production was observed 10 h after induction with methanol (61 U mL(-1)). The fold increase in mannanase production was estimated as approximately 12-fold and approximately 2-fold in A. sojae and P. pastoris, respectively, when compared with A. fumigatus. Both recombinant enzymes showed molecular mass of about 60 kDa and similar specific activities ( approximately 350 U mg(-1) protein). Temperature optima were at 60 degrees C and 45 degrees C, and maximum activity was at pH 4.5 and 5.2 for A. sojae and P. pastoris, respectively. The enzyme from P. pastoris was more stable retaining most of the activity up to 50 degrees C, whereas the enzyme from A. sojae rapidly lost activity above 40 degrees C.

Hydrothermal synthesis of nanostructured TiO2 particles and characterization of their photocatalytic antimicrobial activity.

Nanostructured titania particles were synthesized by using hydrothermal processing and the photocatalytic antimicrobial activities were characterized. Both sol-gel synthesized and commercial TiO2 (anatase) samples were processed with two step hydrothermal treatments, under alkaline and neutral conditions. Scanning Electron Microscope (SEM) images showed that alkaline treatment yields nanofibers and lamellar structured particles from the commercial anatase and sol-gel synthesized samples respectively. Further treatment of nanofibers and nanostructured lamellar particles with distilled water results with crystal growth and the formation of nano structured bipyramidal crystalline particles. The photocatalytic antimicrobial activities of the samples were determined against Escherichia coil under irradiation. It was observed that the samples treated under alkaline conditions have improved activity than the original anatase samples. Limited activity and resulting time lag in bacterial inactivation were observed for hydrothermally treated samples with distilled water. However, a post treatment comprising the UV irradiation in aqueous conditions enhanced the photocatalytic activity.

Purification, characterization, and identification of a novel bifunctional catalase-phenol oxidase from Scytalidium thermophilum.

A novel bifunctional catalase with an additional phenol oxidase activity was isolated from a thermophilic fungus, Scytalidium thermophilum. This extracellular enzyme was purified ca. 10-fold with 46% yield and was biochemically characterized. The enzyme contains heme and has a molecular weight of 320 kDa with four 80 kDa subunits and an isoelectric point of 5.0. Catalase and phenol oxidase activities were most stable at pH 7.0. The activation energies of catalase and phenol oxidase activities of the enzyme were found to be 2.7 +/- 0.2 and 10.1 +/- 0.4 kcal/mol, respectively. The pure enzyme can oxidize o-diphenols such as catechol, caffeic acid, and L-DOPA in the absence of hydrogen peroxide and the highest oxidase activity is observed against catechol. No activity is detected against tyrosine and common laccase substrates such as ABTS and syringaldazine with the exception of weak activity with p-hydroquinone. Common catechol oxidase inhibitors, salicylhydroxamic acid and p-coumaric acid, inhibit the oxidase activity. Catechol oxidation activity was also detected in three other catalases tested, from Aspergillus niger, human erythrocyte, and bovine liver, suggesting that this dual catalase-phenol oxidase activity may be a common feature of catalases.

Production and characterization of films from cotton stalk xylan.

Composite film production based on cotton stalk xylan was studied, and the mechanical and physical properties of the films formed were investigated. Xylan and lignin were separated from cellulose by alkali extraction and, then, lignin was removed using ethanol washing. Self-supporting continuous films could not be produced using pure cotton stalk xylan. However, film formation was achieved using 8-14% (w/w) xylan without complete removal of lignin during xylan isolation. Keeping about 1% lignin in xylan (w/w) was determined to be sufficient for film formation. Films were produced by casting the film-forming solutions, followed by solvent evaporation in a temperature (20 degrees C) and relative humidity (40%) controlled environment. The elastic modulus and hypothetical coating strength of the films obtained by using 8% xylan were significantly different from the ones containing 10-14% xylan. The water vapor transfer rates (WVTR) decreased with increasing xylan concentration, which made the films thicker. The glycerol addition as an additional plasticizer resulting in more stretchable films having higher WVTR and lower water solubility values. As a result, film production was successfully achieved from xylan, which was extracted from an agricultural waste (cotton stalk), and the film-forming effect of lignin on pure xylan has been demonstrated.

Enzymatic production of xylooligosaccharides from cotton stalks.

Xylooligosaccharide (XO) production was performed from xylan, which was obtained by alkali extraction from cotton stalk, a major agricultural waste in Turkey. Enzymatic hydrolysis was selected to prevent byproduct formation such as xylose and furfural. Xylan was hydrolyzed using a commercial xylanase preparation, and the effects of pH, temperature, hydrolysis period, and substrate and enzyme concentrations on the XO yield and degree of polymerization (DP) were investigated. Cotton stalk contains about 21% xylan, the composition of which was determined as 84% xylose, 7% glucose, and 9% uronic acid after complete acid hydrolysis. XOs in the DP range of 2-7 (X6 approximately X5>X2>X3) were obtained with minor quantities of xylose in all of the hydrolysis conditions used. Although after 24 h of hydrolysis at 40 degrees C, the yield was about 53%, the XO production rate leveled off after 8-24 h of hydrolysis. XO yield was affected by all of the parameters investigated; however, none of them affected the DP of the end product significantly, except the hydrolysis period. Enzyme hydrolysis was maintained by the addition of fresh substrate after 72 h of hydrolysis, indicating the persistence of enzyme activity. The optimal hydrolysis conditions were determined as 40 degrees C, pH 5.4, and 2% xylan. The obtained product was fractionated via ultrafiltration by using 10, 3, and 1 kDa membranes. Complete removal of xylanase and unhydrolyzed xylan was achieved without losing any oligosaccharides having DP 5 or smaller by 10 kDa membrane. After a two-step membrane processing, a permeate containing mostly oligosaccharides was obtained.

Drying temperature and relative humidity effects on wheat gluten film properties.

The mechanical and physical properties of glycerol-plasticized wheat gluten films dried at different temperatures (20, 50, and 80 degrees C) and relative humidities (35 and 70% RH) were investigated. Dispersion of wheat gluten was prepared at pH 11 in aqueous solution. Films were obtained by casting the wheat gluten suspension, followed by solvent evaporation in a temperature and relative humidity controlled chamber. Decreasing relative humidity altered most of the mechanical properties. At 35% RH, tensile strength increased when drying temperature increased. However, at 70% RH, tensile strength decreased when temperature increased. Thickness of the films decreased by increasing temperature. Hypothetical coating strength increased with increasing drying temperature at 35% RH. However, at 70% RH, a maximum value was observed at 50 degrees C. Films produced at 80 degrees C exhibited low solubility in aqueous solution. Addition of 1.5% (w/v) sodium dodecyl sulfate increased solubility of all of the films except the film dried at 50 degrees C and 70% RH. Overall, drying temperature and relative humidity affected mechanical and physical properties of the wheat gluten films. However, the effect of drying temperature was more pronounced than the effect of relative humidity.

Use of xylan, an agricultural by-product, in wheat gluten based biodegradable films: mechanical, solubility and water vapor transfer rate properties.

The possibility of using xylan, as an agricultural by-product, for production of composite films in combinations with wheat gluten was investigated. Different levels of xylan (0-40% w/w) were incorporated into wheat gluten to form biodegradable composite films. Films were prepared at pH 4 and 11, and dried at either uncontrolled or controlled conditions. The mechanical properties, solubilities and water vapour transfer rate (WVTR) of the composite films were studied. Films were obtained with added xylan without decreasing film-forming quality. Xylan can be used as an additive, as much as 40% (w/w), in wheat gluten films. Changing pH, wheat gluten/xylan ratio, xylan type and drying conditions affected mechanical and solubility properties, however, WVTR was not affected by xylan additions. Wheat gluten/xylan composite films having different characteristics can be produced depending on xylan type, composition and process conditions.