Lipid production from Yarrowia lipolytica Po1g grown in sugarcane bagasse hydrolysate

This study investigated the possibility of utilizing detoxified sugarcane bagasse hydrolysate (DSCBH) as an alternative carbon source to culture Yarrowia lipolytica Po1g for microbial oil and biodiesel production. Sugarcane bagasse hydrolysis with 2.5% HCl resulted in maximum total sugar concentration (21.38 g/L) in which 13.59 g/L is xylose, 3.98 g/L is glucose, and 2.78 g/L is arabinose. Detoxification of SCBH by Ca(OH)₂ neutralization reduced the concentration of 5-hydroxymethylfurfural and furfural by 21.31% and 24.84%, respectively. Growth of Y. lipolytica Po1g in DSCBH with peptone as the nitrogen source gave maximum biomass concentration (11.42 g/L) compared to NH₄NO₃ (6.49 g/L). With peptone as the nitrogen source, DSCBH resulted in better biomass concentration than d-glucose (10.19 g/L), d-xylose (9.89 g/L) and NDSCBH (5.88 g/L). The maximum lipid content, lipid yield and lipid productivity of Y. lipolytica Po1g grown in DSCBH and peptone was 58.5%, 6.68 g/L and 1.76 g/L-day, respectively.     

Poly-3-hydroxybutyrate (P3HB) production by bacteria from xylose,glucose and sugarcane bagasse hydrolysate

Fifty-five bacterial strains isolated from soil were screened for efficient poly-3-hydroxybutyrate (P3HB) biosynthesis from xylose. Three strains were also evaluated for the utilization of bagasse hydrolysate after different detoxification steps. The results showed that activated charcoal treatment is pivotal to the production of a hydrolysate easy to assimilate. Burkholderia cepacia IPT 048 and B. sacchari IPT 101 were selected for bioreactor studies, in which higher polymer contents and yields from the carbon source were observed with bagasse hydrolysate, compared with the use of analytical grade carbon sources. Polymer contents and yields, respectively, reached 62% and 0.39 g g^–1 with strain IPT 101 and 53% and 0.29 g g^–1 with strain IPT 048. A higher polymer content and yield from the carbon source was observed under P limitation, compared with N limitation, for strain IPT 101. IPT 048 showed similar performances in the presence of either growth-limiting nutrient. In high-cell-density cultures using xylose plus glucose under P limitation, both strains reached about 60 g l^–1 dry biomass, containing 60% P3HB. Polymer productivity and yield from this carbon source reached 0.47 g l^–1 h^–1 and 0.22 g g^–1, respectively.     

Hydrolysis of cellulose derived from steam exploded bagasse by Penicillium cellulases: Comparison with commercial cellulase

A complete cellulase from Penicillium pinophilum was evaluated for the hydrolysis of α-cellulose derived from steam exploded sugarcane bagasse and other cellulosic substrates. α-Cellulose at 1% substrate concentration was completely hydrolyzed by Penicillium cellulase within 3 h wherein at 10% the hydrolysis was 100% within 24 h with an enzyme loading of 10 FPU/g. The hydrolysate yielded glucose as major end product as analyzed by HPLC. Under similar conditions, hydrolysis of Sigmacell (microcrystalline cellulose), CP-123 (pulverized cellulose powder) and ball milled Solka Floc were 42%, 56% and 52%, respectively. Further the hydrolysis performance of Penicillium sp. cellulase is compared with Trichoderma reesei cellulase (Accellerase^TM 1000) from Genencore. The kinetics of hydrolysis with respect to enzyme and substrate concentration will be presented.     

Secretome analysis of Trichoderma reesei and Aspergillus niger cultivated by submerged and sequential fermentation processes: Enzyme production for sugarcane bagasse hydrolysis

Cellulases and hemicellulases from Trichoderma reesei and Aspergillus niger have been shown to be powerful enzymes for biomass conversion to sugars, but the production costs are still relatively high for commercial application. The choice of an effective microbial cultivation process employed for enzyme production is important, since it may affect titers and the profile of protein secretion. We used proteomic analysis to characterize the secretome of T. reesei and A. niger cultivated in submerged and sequential fermentation processes. The information gained was key to understand differences in hydrolysis of steam exploded sugarcane bagasse for enzyme cocktails obtained from two different cultivation processes. The sequential process for cultivating A. niger gave xylanase and β-glucosidase activities 3- and 8-fold higher, respectively, than corresponding activities from the submerged process. A greater protein diversity of critical cellulolytic and hemicellulolytic enzymes were also observed through secretome analyses. These results helped to explain the 3-fold higher yield for hydrolysis of non-washed pretreated bagasse when combined T. reesei and A. niger enzyme extracts from sequential fermentation were used in place of enzymes obtained from submerged fermentation. An enzyme loading of 0.7 FPU cellulase activity/g glucan was surprisingly effective when compared to the 5–15 times more enzyme loadings commonly reported for other cellulose hydrolysis studies. Analyses showed that more than 80% consisted of proteins other than cellulases whose role is important to the hydrolysis of a lignocellulose substrate. Our work combined proteomic analyses and enzymology studies to show that sequential and submerged cultivation methods differently influence both titers and secretion profile of key enzymes required for the hydrolysis of sugarcane bagasse. The higher diversity of feruloyl esterases, xylanases and other auxiliary hemicellulolytic enzymes observed in the enzyme mixtures from the sequential fermentation could be one major reason for the more efficient enzyme hydrolysis that results when using the combined secretomes from A. niger and T. reesei.     

Fermentation of pretreated sugarcane bagasse hemicellulose hydrolysate to ethanol by <Emphasis Type="Italic">Pachysolen tannophilus</Emphasis>

Sugarcane bagasse hemicellulose hydrolysates, pretreated by either over-liming or electrodialysis and, supplemented with nutrient materials, were fermented to ethanol using Pachysolen tannophilus DW06. Compared with detoxification by over-liming, detoxification by electrodialysis decreased the loss of sugar and increased the acetic acid removal, leading to better fermentability. A batch culture with electrodialytically pretreated hydrolysate as substrate was developed giving 21 g ethanol l⁻¹ with a yield of 0.35 g g⁻¹ sugar and productivity of 0.59 g l⁻¹ h⁻¹.     

The influence of pH and dilution rate on continuous production of xylitol from sugarcane bagasse hemicellulosic hydrolysate by C. guilliermondii

Continuous fermentation of sugarcane bagasse hemicellulosic hydrolysate by the yeast Candida guilliermondii FTI 20037 was used for xylitol production from xylose. Experiments were carried out in a reactor with 1.25 l of treated hydrolysate, at 30 °C and 300 rpm. A 2² full-factorial central composite design was employed for experimental study and analysis of the results. A statistical analysis of the results showed that the effects of the pH and dilution rate (D), the interactions between these variables and the second-order effect of D on the xylitol volumetric productivity (Q_p) were significant at a 95% confidence level. The second-order effect of pH was also significant at a 90% confidence level. The k_La effect on the Q_p was not significant. A volumetric productivity of 0.68 g/l h, representing 95.8% of the predicted value (0.72 g/l h), was obtained.     

Adsorption of Cu(II), Cd(II), and Pb(II) from aqueous single metal solutions by sugarcane bagasse and mercerized sugarcane bagasse chemically modified with succinic anhydride

This work describes the preparation of new chelating material from mercerized sugarcane bagasse. The first part treats the chemical modification of non-mercerized sugarcane bagasse (SCB) and twice-mercerized sugarcane bagasse (MMSCB) with succinic anhydride. Mass percent gains (mpg) and degrees of succinylation (DS) of succinylated non- and twice-mercerized sugarcane bagasse 1 (SCB 1 and MMSCB 1) were calculated. MMSCB 1 exhibited an increase in mpg and DS of 49.2% and 0.9 mmol/g in relation to SCB 1. SCB 2 and MMSCB 2 were obtained by treatment of MMSCB 1 and SCB 1 with bicarbonate solution to release the carboxylate functions and characterized by FTIR. The second part evaluates and compares the adsorption capacity of SCB 2 and MMSCB 2 for Cu²⁺, Cd²⁺ and Pb²⁺ ions in an aqueous single metal solution. Adsorption isotherms were developed using Langmuir model. MMSCB 2 exhibited an increase in Q_max for Cd²⁺ (43.6 mg/g) and Pb²⁺ (83.3 mg/g) in relation to SCB 2.     

Kinetics of the solid state fermentation of sugarcane bagasse by Thermoascus aurantiacus for the production of xylanase

Xylanase was produced by solid-state fermentation using Thermoascus aurantiacus. Maximum production (500 U g^–1 bagasse) was achieved on the sixth day of cultivation on solid sugarcane bagasse medium supplemented with 15% (v/w) rice bran extract. The fungal biomass, determined from its glucosamine content, reached 28 mg g^–1 on the 8th day of cultivation. The cell yield against O₂ (Y _x/o =0.18g _cell/gO₂) and maintenance coefficient (m ₀=0.013g _O2/g _cell h) were determined with the low Y _x/o value for T. aurantiacus agreeing with the calculated value.     

Batch xylitol production by<Emphasis Type="Italic"> Candida guilliermondii</Emphasis> FTI 20037 from sugarcane bagasse hemicellulosic hydrolyzate at controlled pH values

Batch fermentation of sugarcane bagasse hemicellulosic hydrolyzate by the yeast Candida guilliermondii FTI 20037 was performed using controlled pH values (3.5, 5.5, 7.5). The maximum values of xylitol volumetric productivity (Q _p=0.76 g/l h) and xylose volumetric consumption (Q _s=1.19 g/l h) were attained at pH 5.5. At pH 3.5 and 7.5 the Q _p value decreased by 66 and 72%, respectively. Independently of the pH value, Y _x/s decreased with the increase in Y _p/s suggesting that the xylitol bioconversion improves when the cellular growth is limited. At the highest pH value (7.5), the maximum specific xylitol production value was the lowest (q _pmax=0.085 g/l h.), indicating that the xylose metabolism of the yeast was diverted from xylitol formation to cell growth.List of symbols P _max xylitol concentration (g/l) - Q _x volumetric cell production rate (g/l h) - Q _s volumetric xylose uptake rate (g/l h) - Q _p volumetric xylitol production rate (g/l h) - q _pmax specific xylitol production (g/g h) - q _smax specific xylose uptake rate (g/g h) - _max specific cell growth rate (h^–1) - Y _p/s xylitol yield coefficient, g xylitol per g xylose consumed (g/g) - Y _p/x xylitol yield coefficient, g xylitol per g dry cell mass produced (g/g) - Y _x/s cell yield coefficient, g dry cell mass per g xylose consumed (g/g) - _cell percentage of the cell yield from the theoretical value (%) - _xylitol percentage of xylitol yield from the theoretical value (%)     

Biogas upgrading using single-walled carbon nanotubes by molecular simulation

Abstract

Biogas from anaerobic digestion of biological wastes is a renewable energy resource that mainly contains CH₄, CO₂, trace amounts of H₂S and a fraction of H₂O vapour. In order to transfer biogas into biomethane to meet the standards for use as vehicle fuel or for injection in the natural gas grid, removing H₂S from biogas in advance is necessary. In addition, biogas is usually saturated with water vapour. It is significant to study the effect of the presence of H₂O on the biogas separation performance. Adsorption of H₂S/CO₂/CH₄ and H₂O/CO₂/CH₄ ternary mixtures using single-walled carbon nanotubes (SWCNT) were investigated via the Grand Canonical Monte Carlo (GCMC) method. We studied the effects of carbon nanotube diameter, –COOH modification, temperature and pressure on H₂S adsorption. The results indicate that the presence of hydrophilic –COOH groups does affect the separation of H₂S/CO₂/CH₄ mixtures. Temperature swing adsorption is more suitable than pressure swing adsorption for the separation of H₂S/CO₂/CH₄ mixtures. The effect of water vapour on the separation of CO₂/CH₄ was also investigated. The result shows that the presence of H₂O has little effect on the selectivity of CO₂/CH₄ in pristine CNT, but the selectivity of CO₂/CH₄ with the presence of H₂O is markedly enhanced after modification in –COOH modified SWCNT with specific modification degree. It is expected that this work could provide some useful information for biogas upgrading.     

Production and characterization of cellulolytic and xylanolytic enzymes from the ligninolytic white-rot fungus Phanerochaete chrysosporium grown on sugarcane bagasse

The ligninolytic white-rot fungus Phanerochaete chrysosporium BKM-F-1767 produced extracellular cellulolytic enzymes (carboxymethylcellulase, CMCase and -glucosidase) and xylanolytic enzymes (xylanase and -xylosidase) in liquid medium containing 1.0% sugarcane bagasse with or without 1.0% glucose. The changes in pH and soluble protein content were monitored in the culture filtrates. The results obtained showed that the pH decreased after 3 days and then increased. The soluble protein content increased and reached the maximum value after 12 days. The results showed that the activities of enzymes were higher in the case of sugarcane bagasse without glucose. The characterization study indicated that the optimum pH values were 4.6, 4.2, 5.0 and 5.0 for CMCase, -glucosidase, xylanase and -xylosidase, respectively and the optimum temperatures were 60, 70, 65 and 60 °C for the investigated enzymes, respectively. The results showed also that after prolonged heating (5 h) at 60 °C, CMCase, -glucosidase, xylanase and -xylosidase retained 81.2, 86.8, 51.5 and 27.4% activity, respectively.     

Structure of the extracellular glutathione S-transferase OvGST1 from the human pathogenic parasite Onchocerca volvulus

Onchocerciasis or river blindness, caused by the filarial worm Onchocerca volvulus, is the world’s second leading infectious cause of blindness. In order to chronically infect the host, O. volvulus has evolved molecular strategies that influence and direct immune responses away from the modes most damaging to it. The O. volvulus GST1 (OvGST1) is a unique glutathione S-transferase (GST) in that it is a glycoprotein and possesses a signal peptide that is cleaved off in the process of maturation. The mature protein starts with a 25-amino-acid extension not present in other GSTs. In all life stages of the filarial worm, it is located directly at the parasite-host interface. Here, the OvGST1 functions as a highly specific glutathione-dependent prostaglandin D synthase (PGDS). The enzyme therefore has the potential to participate in the modulation of immune responses by contributing to the production of parasite-derived prostanoids and restraining the host’s effector responses, making it a tempting target for chemotherapy and vaccine development. Here, we report the crystal structure of the OvGST1 bound to its cofactor glutathione at 2.0 Å resolution. The structure reveals an overall structural homology to the haematopoietic PGDS from vertebrates but, surprisingly, also a large conformational change in the prostaglandin binding pocket. The observed differences reveal a different vicinity of the prostaglandin H₂ binding pocket that demands another prostaglandin H₂ binding mode to that proposed for the vertebrate PGDS. Finally, a putative substrate binding mode for prostaglandin H₂ is postulated based on the observed structural insights.     

Molecular mechanisms of nitrogen dioxide induced epithelial injury in the lung

The lung can be exposed to a variety of reactive nitrogen intermediates through the inhalation of environmental oxidants and those produced during inflammation. Reactive nitrogen species (RNS) include, nitrogen dioxide (·NO₂) and peroxynitrite (ONOO^–). Classically known as a major component of both indoor and outdoor air pollution, ·NO₂ is a toxic free radical gas. ·NO₂ can also be formed during inflammation by the decomposition of ONOO^– or through peroxidase-catalyzed reactions. Due to their reactive nature, RNS may play an important role in disease pathology. Depending on the dose and the duration of administration, ·NO₂ has been documented to cause pulmonary injury in both animal and human studies. Injury to the lung epithelial cells following exposure to ·NO₂ is characterized by airway denudation followed by compensatory proliferation. The persistent injury and repair process may contribute to airway remodeling, including the development of fibrosis. To better understand the signaling pathways involved in epithelial cell death by ·NO₂ or other RNS, we routinely expose cells in culture to continuous gas-phase ·NO₂. Studies using the ·NO₂ exposure system revealed that lung epithelial cell death occurs in a density dependent manner. In wound healing experiments, ·NO₂ induced cell death is limited to cells localized in the leading edge of the wound. Importantly, ·NO₂-induced death does not appear to be dependent on oxidative stress per se. Potential cell signaling mechanisms will be discussed, which include the mitogen activated protein kinase, c-Jun N-terminal Kinase and the Fas/Fas ligand pathways. During periods of epithelial loss and regeneration that occur in diseases such as asthma or during lung development, epithelial cells in the lung may be uniquely susceptible to death. Understanding the molecular mechanisms of epithelial cell death associated with the exposure to ·NO₂ will be important in designing therapeutics aimed at protecting the lung from persistent injury and repair.     

A study of gramicidin using deuterium oxide

The single channel conductivity of the gramicidin channel has been measured for all the alkali ions using both H₂O and ²H₂O as a medium. Significant changes in conductivity with medium have been observed in all cases except lithium.     

Involvement of phospholipid, biomembrane integrity, and NO peroxidase activity in the NO catabolism by cytochrome c oxidase

The physiological regulation of mitochondrial respiration by NO has been reported to result from the reversible binding of NO to the two-electron reduced binuclear center (Fe(2+)(a3)-Cu(1+)(B)) of cytochrome c oxidase (CcO). Although the role of CcO and its derived catalytic intermediates in the catabolism of NO has been documented, little has been established for the enzyme in its fully oxidized state (Fe(3+)(a3)-Cu(2+)(B)). We report: (1) CcO, in its fully oxidized state, represents the major component of the mitochondrial electron transport chain for NO consumption as controlled by the binding of NO to its binuclear center. Phospholipid enhances NO consumption by fully oxidized CcO, whereas the consumption of NO is slowed down by membrane structure and membrane potential when CcO is embedded in the phospholipid bilayer. (2) In the presence of H(2)O(2), CcO was shown to serve as a mitochondria-derived NO peroxidase. A CcO-derived protein radical intermediate was induced and involved in the modulation of NO catabolism.     

Visible light-induced H2 production from cellulose using photosensitization of Mg chlorophyll a

A photoinduced-H₂ production system, coupling cellulose degradation by cellulase and glucose dehydrogenase (GDH) and H₂ production with colloidal Pt as a catalyst using the visible light-induced photosensitization of Mg chlorophyll a, has been developed. When the sample solution containing methylcellulose, cellulase, GDH, NAD⁺, Mg chlorophyll a, Methyl viologen and colloidal Pt was irradiated, continuous H₂ production was observed. The amount of H₂ production was about 12 mol after 4 h irradiation.     

Isotopic signatures of N2O produced by ammonia-oxidizing archaea from soils

N₂O gas is involved in global warming and ozone depletion. The major sources of N₂O are soil microbial processes. Anthropogenic inputs into the nitrogen cycle have exacerbated these microbial processes, including nitrification. Ammonia-oxidizing archaea (AOA) are major members of the pool of soil ammonia-oxidizing microorganisms. This study investigated the isotopic signatures of N₂O produced by soil AOA and associated N₂O production processes. All five AOA strains (I.1a, I.1a-associated and I.1b clades of Thaumarchaeota) from soil produced N₂O and their yields were comparable to those of ammonia-oxidizing bacteria (AOB). The levels of site preference (SP), δ¹⁵N^bulk and δ¹⁸O -N₂O of soil AOA strains were 13–30%, −13 to −35% and 22–36%, respectively, and strains MY1–3 and other soil AOA strains had distinct isotopic signatures. A ¹⁵N-NH₄⁺-labeling experiment indicated that N₂O originated from two different production pathways (that is, ammonia oxidation and nitrifier denitrification), which suggests that the isotopic signatures of N₂O from AOA may be attributable to the relative contributions of these two processes. The highest N₂O production yield and lowest site preference of acidophilic strain CS may be related to enhanced nitrifier denitrification for detoxifying nitrite. Previously, it was not possible to detect N₂O from soil AOA because of similarities between its isotopic signatures and those from AOB. Given the predominance of AOA over AOB in most soils, a significant proportion of the total N₂O emissions from soil nitrification may be attributable to AOA.     

A comprehensive study on the contents and leaching of trace elements from fly-ash originating from polish hard coal by NAA and AAS methods

In order to assess the environmental risks associated with the emission of fly-ash into the atmosphere and its storage on waste heaps, the trace element contents of fly-ashes from burning Polish hard coal were determined by a newly developed INAA method. Leaching of trace elements from the fly-ash by water and H₂SO₄ solution (pH≈2.5) simulating acid rain, respectively, was studied using AAS and spectrophotometric methods. Analogous experiments were done with neutron-irradiated fly-ash, following the composition of the eluate gamma-spectrometrically. The new fine fly-ash (CTA-FFA-1) candidate reference material was prepared, and the certification was undertaken on the basis of an international intercomparison run. Preliminary evaluation of results shows that at least 38 elements will be certified and, in addition, the “information values” for at least 12 elements will be given.     

Removal of Ca(II) and Mg(II) from aqueous single metal solutions by mercerized cellulose and mercerized sugarcane bagasse grafted with EDTA dianhydride (EDTAD)

In a previous work, chemically modified cellulose (EMC) and sugarcane bagasse (EMMB) were prepared from mercerized cellulose (MC) and twice-mercerized sugarcane bagasse (MMB) using ethylenediaminetetraacetic dianhydride (EDTAD) as modifying agent. In this work we described in detail the modification of these materials in function of reaction time and EDTAD amount in the reaction media. The resistance of ester bond at pH 1, 2, 11, and 12 was also evaluated by FTIR. The results were used to model the hydrolysis process and a kinetic model was proposed. The modified materials (EMMB and EMC) were used to adsorb Ca²⁺ and Mg²⁺ ions from aqueous single solutions. The adsorption isotherms were developed at two pH values. These materials showed maximum adsorption capacities for Ca²⁺ and Mg²⁺ ions ranging from 15.6 to 54.1 mg/g and 13.5 to 42.6 mg/g, respectively. The modified material from sugarcane bagasse (EMMB) showed larger maximum adsorption capacities than modified material from cellulose (EMC) for both metals.