Single-Cell Protein Production in Alkaline Mesquite Wood Extracts

A nine-member, mixed, cellulolytic, bacterial culture was used to evaluate the effects of sodium hydroxide normality, length and temperature of treatment, and the ratio of volume of alkali to mesquite wood on the suitability of alkali mesquite wood extractives as nutrients for bacterial growth. The presence or absence of air during the extraction process did not significantly affect results. The amount of lignin extracted and the total loss in weight of the wood during extraction were correlated to both alkali concentration and temperature. Neutralized extracts supported bacterial growth; growth was inversely related to the final salt concentration of the neutralized extract. Deionized extracts were superior to acid-neutralized extracts for the support of bacterial growth. The optimum conditions for extraction were 2.5 N NaOH at 30°C for 12 h. The study demonstrated that nutrients as well as growth inhibitory compounds are released from wood by alkali treatment. This study of alkali wood extracts and the previous study of washed alkali treated wood residues provide a data base for the optimization of alkali treatments of hard woods that are to be used as nutrients for the growth of cellulolytic cultures.     

Investigation on catechin as a beech wood decay biomarker

A high-performance liquid chromatography (HPLC) method based on the evolution of wood extractives was developed to follow the first stages of fungal degradation of beech wood exposed to Coriolus versicolor. The nature and the quantity of the extracts initially present in wood depended on the extraction conditions and also on the wood-drying conditions (time and temperature). The most interesting fraction was soxhlet extracted with acetone at 56 °C for 6 h. The best conditions to avoid extractives degradation consisted of a moderate drying at 55 °C for 48 h allowing identification of catechin as potential tracer. After 2 weeks of wood blocks exposure to C. versicolor, analysis of their acetonic extractives showed that catechin signal initially detected in beech wood, had totally disappeared. Treatment of wood with an appropriate fungicide such as propiconazole before exposure to C. versicolor, prevents the catechin amount from any variation. The comparison of these results with the classical weight loss (WL) measurements obtained after long-time experiments on treated and untreated wood blocks shows that the catechin amount evolution, monitored during 2 weeks, correlates with the wood resistance evaluated during 16 weeks, allowing the use of this flavonoid as a valuable biomarker of wood decay.     

Isolation of the extractives and betulin from birch bark exposed to a microwave field

The process of isolation of the extractives and betulin from birch bark exposed to a microwave field was investigated. It was shown that under the use of microwave treatment in a microwave field the duration of the extraction process was reduced 10–15 times, as compared with the extraction by infusion. The influence of the main parameters on the process of microwave extraction of extractives from birch bark was studied. Using high performance liquid chromatography we established quantitatively the content of betulin in the birch bark extracts.     

Evaluation of the decay resistance properties of Cerbera odollam extracts and their influence on properties of particleboard

Metallic-based wood preservatives currently face some restrictions over disposal and environmental issues; one possibility to develop new more benign systems is to study extractives in naturally durable woody plants. This study investigated the resistance of extracts from the leaf, fruit, wood, bark, seed and flower of Cerbera odollam to deterioration from fungus and termites. Antifungal assays with n-hexane, ethyl acetate, ethanol and methanol extracts were evaluated using paper discs. Termite mortality was evaluated with the methanol extract against Coptotermes gestroi. Physical and protective properties of particleboard impregnated with C. odollam extracts, including thickness swelling, internal bond strength, formaldehyde release, termite-decay and soil burial decay were investigated. Methanol wood extracts from C. odollam showed the highest activities against Trametes versicolor, Pycnoporus sanguineus, and Schizophyllum commune in the paper disc antifungal assay. Methanol flower extracts exhibited high performance in termite mortality, termite-decay and soil burial decay. Thickness swelling, internal bond strength and the formaldehyde emission of particleboard specimens treated with methanol extracts of C. odollam were up to the EN Standards.     

A sustainable woody biomass biorefinery

Woody biomass is renewable only if sustainable production is imposed. An optimum and sustainable biomass stand production rate is found to be one with the incremental growth rate at harvest equal to the average overall growth rate. Utilization of woody biomass leads to a sustainable economy. Woody biomass is comprised of at least four components: extractives, hemicellulose, lignin and cellulose. While extractives and hemicellulose are least resistant to chemical and thermal degradation, cellulose is most resistant to chemical, thermal, and biological attack. The difference or heterogeneity in reactivity leads to the recalcitrance of woody biomass at conversion. A selection of processes is presented together as a biorefinery based on incremental sequential deconstruction, fractionation/conversion of woody biomass to achieve efficient separation of major components. A preference is given to a biorefinery absent of pretreatment and detoxification process that produce waste byproducts. While numerous biorefinery approaches are known, a focused review on the integrated studies of water-based biorefinery processes is presented. Hot-water extraction is the first process step to extract value from woody biomass while improving the quality of the remaining solid material. This first step removes extractives and hemicellulose fractions from woody biomass. While extractives and hemicellulose are largely removed in the extraction liquor, cellulose and lignin largely remain in the residual woody structure. Xylo-oligomers, aromatics and acetic acid in the hardwood extract are the major components having the greatest potential value for development. Higher temperature and longer residence time lead to higher mass removal. While high temperature (>200°C) can lead to nearly total dissolution, the amount of sugars present in the extraction liquor decreases rapidly with temperature. Dilute acid hydrolysis of concentrated wood extracts renders the wood extract with monomeric sugars. At higher acid concentration and higher temperature the hydrolysis produced more xylose monomers in a comparatively shorter period of reaction time. Xylose is the most abundant monomeric sugar in the hydrolysate. The other comparatively small amounts of monomeric sugars include arabinose, glucose, rhamnose, mannose and galactose. Acetic acid, formic acid, furfural, HMF and other byproducts are inevitably generated during the acid hydrolysis process. Short reaction time is preferred for the hydrolysis of hot-water wood extracts. Acid hydrolysis presents a perfect opportunity for the removal or separation of aromatic materials from the wood extract/hydrolysate. The hot-water wood extract hydrolysate, after solid-removal, can be purified by Nano-membrane filtration to yield a fermentable sugar stream. Fermentation products such as ethanol can be produced from the sugar stream without a detoxification step.     

Comparison of methods to evaluate the potential of fungal growth on decay of spruce wood after short-time treatment

Several analytical methods were compared to evaluate characteristic wood decaying fungi for their potential to depolymerise lignin on spruce wood particles. Wood samples were treated with the white rot fungi Phlebia brevispora, Ceriporiopsis subvermispora, Merulius tremellosus, Pycnoporus sanguineus, Trametes pubescens and with the brown rot fungus Gloeophyllum trabeum. The UV absorbancies of crude ethanol extracts, total extractives content from sequential extraction, ligninolytic enzyme activities, lignin solubilisation and decrease of lignin content were compared. It was shown, that, in early decay stages, UV absorbancies of crude ethanol extracts and total extractives content correlate well with lignin degradation, increase of acid soluble lignin and increased production of ligninolytic enzymes (total peroxidase). Lignin content was determined using FT-NIR spectroscopy as well as by wet-chemical analysis, indicating a very good correlation between the two methods. According to the different analytical methods, the tested fungi can be classified into three categories based on their characteristic behaviour: brown rot, “slow” and “fast” white rot.     

The GSTome Reflects the Chemical Environment of White-Rot Fungi

White-rot fungi possess the unique ability to degrade and mineralize all the different components of wood. In other respects, wood durability, among other factors, is due to the presence of extractives that are potential antimicrobial molecules. To cope with these molecules, wood decay fungi have developed a complex detoxification network including glutathione transferases (GST). The interactions between GSTs from two white-rot fungi, Trametes versicolor and Phanerochaete chrysosporium, and an environmental library of wood extracts have been studied. The results demonstrate that the specificity of these interactions is closely related to the chemical composition of the extracts in accordance with the tree species and their localization inside the wood (sapwood vs heartwood vs knotwood). These data suggest that the fungal GSTome could reflect the chemical environment encountered by these fungi during wood degradation and could be a way to study their adaptation to their way of life.     

Effects of bacterial treatments on wood extractives

Bacterial strains were isolated from spruce wood chips and their ability to reduce the content of wood extractives was studied. Strains were screened by cultivation on liquid media containing wood extractives as the major nutrient. Some bacterial species could decrease remarkably the amount of extractives in the liquid media and reduced the amount of triglycerides, steryl esters and total extractives by 100, 20 and 39%, respectively. Spruce wood chips were treated in controlled conditions with selected bacteria to test their effects on the chips. All the bacteria grew well on wood chips. The effect of bacterial metabolism on wood extractives was significant. Bacterial treatments reduced the amount of lipophilic extractives by 16-38% in 1 week of treatment and up to 67% in 2 weeks. The most efficient strain removed 90, 66 and 50% of triglycerides, steryl esters and resin acids, respectively, in 2 weeks. These results indicate that bacteria may be promising agents for the removal of extractives for improved pulping and papermaking processes.     

Fixation,leachability, and decay resistance of wood treated with some commercial extracts and wood preservative salts

Beech (Fagus orientalis) and Scots pine (Pinus sylvestris) wood blocks were treated with some commercial extracts as well as water-based wood preservative salts at various concentrations to increase retention and fixation. The penetration, fixation, and antifungal properties of different treatment solutions were compared with statistical analysis. Retention levels of solutions were generally higher for Scots pine wood than beech wood. The highest retention levels were seen in wood treated with sumac leaf extract and oak valonia extract. Leaching tests indicated that both wood types treated with sumac extracts showed higher retention levels than wood treated with the other fruit and bark extract solutions. Adding 1% water-based wood preservative salts to valonia and sumac leaf extracts increased the retention levels. Concentrations of more than 1% did not contribute to retention either individually or with salt additions. Three percent and higher concentrations of wood-preserving salts accelerated and increased the amount of leaching. The results showed that the extract alone was resistant to leaching. Mycological tests showed that all extractives were significantly effective against wood decay.     

Bacterial biodegradation of extractives and patterns of bordered pit membrane attack in pine wood

Wood extractives, commonly referred to as pitch, cause major problems in the manufacturing of pulp and paper. Treatment of nonsterile southern yellow pine chips for 14 days with Pseudomonas fluorescens, Pseudomonas sp., Xanthomonas campestris, and Serratia marcescens reduced wood extractives by as much as 40%. Control treatments receiving only water lost 11% of extractives due to the growth of naturally occurring microorganisms. Control treatments were visually discolored after the 14-day incubation, whereas bacterium-treated wood chips were free of dark staining. Investigations using P. fluorescens NRRL B21432 showed that all individual resin and fatty acid components of the pine wood extractives were substantially reduced. Micromorphological observations showed that bacteria were able to colonize resin canals, ray parenchyma cells, and tracheids. Tracheid pit membranes within bordered pit chambers were degraded after treatment with P. fluorescens NRRL B21432. P. fluorescens and the other bacteria tested appear to have the potential for biological processing to substantially reduce wood extractives in pine wood chips prior to the paper making process so that problems associated with pitch in pulp mills can be controlled.     

The effect of different solvents and number of extraction steps on the polyphenol content and antioxidant capacity of basil leaves (Ocimum basilicum L.) extracts

The objectives of this study were to determine best conditions for the extraction of phenolic compounds from fresh, frozen and lyophilized basil leaves. The acetone mixtures with the highest addition of acetic acid extracted most of the phenolic compounds when fresh and freeze-dried material have been used. The three times procedure was more effective than once shaking procedure in most of the extracts obtained from fresh basil leaves – unlike the extracts derived from frozen material. Surprisingly, there were not any significant differences in the content of phenolics between the two used procedures in the case of lyophilized basil leaves used for extraction. Additionally, the positive correlation between the phenolic compounds content and antioxidant activity of the studied extracts has been noted. It is concluded that the acetone mixtures were more effective than the methanol ones for polyphenol extraction. The number of extraction steps in most of the cases was also a statistically significant factor affecting the yield of phenolic extraction as well as antioxidant potential of basil leaf extracts.     

Comparison of different procedures for the lipid extraction from HL-60 cells: a MALDI-TOF mass spectrometric study

A human leukaemia cell line--HL-60--can be differentiated into neutrophils or macrophages and both differentiation processes are accompanied by changes of the lipid composition. Various methods were described for the extraction of lipids from cellular systems, but only two of them were applied to the HL-60 cell line so far. In this study we compared five selected extraction methods for the lipid extraction from HL-60 cells with regard to their qualitative analysis by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS): chloroform/methanol at volume ratios 2:1 and 1:2, isopropanol/ chloroform, isopropanol/hexane and butanol. In addition, the cholesterol and phospholipid concentrations in organic extracts were measured by colorimetric assays. Results can be summarized as follows: For the analysis of polar phospholipids obtained from HL-60 cells by MALDI-TOF MS, a chlorofom/methanol (1:2) or isopropanol/chloroform mixture or butanol can be applied as extraction systems On the other hand, if one would like to analyze changes in triacylglycerols, then chloroform/methanol (2:1) would be the method of choice.     

An evaluation of the use of Sep-Pak C18 cartridges for the extraction of vitamin D3 and some of its metabolites from plasma and urine

The use of Sep-Pak C18 cartridges for the extraction of vitamin D and some of its metabolites from plasma and urine has been evaluated by studying the recovery of added tritiated secosteroids. The preparation of the cartridges, recoveries, extraction and elution with a number of solvents, effect of varying flow rates for application and elution, and the effect of increasing volumes of plasma and urine have been investigated. Two methods for the application of secosteroids present in plasma to Sep-Pak C18 cartridges have been examined, using methyl cyanide extracts removing precipitated protein by centrifugation, and using acidified methanolic plasma. Methyl cyanide extracts applied to Sep-Pak C18 cartridges and eluted with methanol or methyl cyanide gave the cleanest extracts suitable for direct HPLC. Acidified methanolic plasma, applied to Sep-Pak C18 cartridges and eluted with methanol or methyl cyanide gave extracts which could not be applied directly to an HPLC--further fractionation using Sep-Pak SIL cartridges was necessary. Recoveries of added tritiated secosteroids using both methods were greater than 80% with the exception of vitamin D itself which was poorly recovered--methyl cyanide extraction giving only 30% recovery and use of acidified methanolic plasma giving 66% recovery.     

Investigation of the pharmaceutical and pharmacological equivalence of different Hawthorn extracts

Seven Hawthorn extracts were tested in isolated guinea pig aorta rings. The effect on noradrenaline- (10 microM) induced contraction was investigated. The extracts were prepared using ethanol (40 to 70% v/v), methanol (40 to 70% v/v), and water as the extraction solvents. The aqueous-alcoholic extracts displayed similar spectra of constituents. They were characterised by similar procyanidin, flavonoid, total vitexin and total phenols content and by similar TLC fingerprint chromatograms. The aqueous extract, however, showed a different fingerprint and a noticeably lower concentration of procyanidins, flavonoids and total phenols but a similar total vitexin content. All 7 extracts had a relaxant effect on the aorta precontracted by noradrenaline and led to relaxations to 44 until 29% of the initial values. The EC50 values of the aqueous-alcoholic extracts varied between 4.16 and 9.8 mg/l. The aqueous extract produced a similarly strong maximal relaxation as the other extracts, but the EC50, at 22.39 mg/l, was markedly higher. The results show that Hawthorn extracts with comparable quality profiles were obtained by using aqueous-alcoholic extraction solvents (40 to 70% ethanol or methanol). The extracts exerted comparable pharmacological effects. When using water as the extraction solvent, both, the spectrum of constituents and the pharmacological effect, deviated remarkably. It is thus possible to obtain bioequivalent extracts with comparable effect profiles by using 40 to 70% ethanol or methanol as the extraction solvent.     

Plumbagin recovery from field specimens of Drosophyllum lusitanicum (L.) link

The naphthoquinone plumbagin has a broad spectrum of biological activities. The aim of this study was to investigate the efficiency of two extraction methods (Soxhlet and ultrasound-assisted extraction) and three solvents (methanol, chloroform and hexane) to recover plumbagin from fresh and dried tissues of field specimens of Drosophyllum lusitanicum (L.) Link. The highest extraction yields were obtained with methanol as solvent and using fresh plant material. The obtained extracts were analysed by gas chromatography with mass spectrometric detection and plumbagin was the major compound present. Plumbagin was quantified in the extracts using the external standard methodology. The results obtained showed that the best recoveries of plumbagin were attained using fresh plant material and there were no significant differences between Soxhlet and ultrasound-assisted extraction. Moreover, hexane proved to be the more appropriate solvent for the extraction of plumbagin, providing high recoveries and the most concentrated extracts, yielding 2.42 mg of plumbagin per gram of plant material with the highest degree of purity. This method is a simple and efficient one to extract large amounts of plumbagin from D. lusitanicum field specimens.     

The effect of water soluble Scots pine (Pinus sylvestris L.) and Sitka spruce [Picea sitchensis (Bong.) carr.] heartwood and sapwood extracts on the growth of selected Trichoderma species

Extractable materials from some timber species have been identified which prevent wood decay; however, little has been reported on the effect(s) of such materials against mould species that colonize timber. With increasing interest in the use of Trichoderma species, both as agents of permeability enhancement and biological control, more information is required on how chemical components within fresh and processed timber influence growth of Trichoderma. Fresh and dried samples of Scots pine and Sitka spruce sapwood and heartwood were leached in a Soxhlet apparatus and the resulting extract was combined with malt extract agar and inoculated with Trichoderma. Trichoderma isolates were inhibited to varying degrees by extractives removed from fresh and dried heartwood of the two timbers. Growth on sapwood extractives, however, showed a lesser degree of inhibition. The implications of the results for applications of Trichoderma in timber are discussed.     

Naturally durable heartwood: evidence for a proposed dual defensive function of the extractives

We previously proposed that extractives in highly durable heartwood may protect wood against fungal colonization and subsequent degradation by dual mechanisms: the extractives have some fungicidal activity and are also free radical scavengers (antioxidants). In short-term laboratory decay tests using two different wood species and decay fungi, the antioxidant 2,6-dimethyl-di-tert-butyl-4-methylphenol (BHT) alone had little or no preservative effect. In contrast, the combination of BHT with different organic commercial biocides always showed an increase in efficacy compared to the organic biocide alone. Consequently, we conclude that the combination of a commercial antioxidant and biocide is synergistic. This implies that extractives may protect wood by more than simply being fungicidal.     

Novel gene expression profiles define the metabolic and physiological processes characteristic of wood and its extractive formation in a hardwood tree species,Robinia pseudoacacia

Wood is of critical importance to humans as a primary feedstock for biofuel, fiber, solid wood products, and various natural compounds including pharmaceuticals. The trunk wood of most tree species has two distinctly different regions: sapwood and heartwood. In addition to the major constituents, wood contains extraneous chemicals that can be removed by extraction with various solvents. The composition and the content of the extractives vary depending on such factors as, species, growth conditions, and time of year when the tree is cut. Despite the great commercial and keen scientific interest, little is known about the tree-specific biology of the formation of heartwood and its extractives. In order to gain insight on the molecular regulations of heartwood and its extractive formation, we carried out global examination of gene expression profiles across the trunk wood of black locust (Robinia pseudoacacia L.) trees. Of the 2,915 expressed sequenced tags (ESTs) that were generated and analyzed in the current study, 55.3% showed no match to known sequences. Cluster analysis of the ESTs identified a total of 2278 unigene sets, which were used to construct cDNA microarrays. Microarray hybridization analyses were then performed to survey the changes in gene expression profiles of trunk wood. The gene expression profiles of wood formation differ according to the region of trunk wood sampled, with highly expressed genes defining the metabolic and physiological processes characteristic of each region. For example, the gene encoding sugar transport had the highest expression in the sapwood, while the structural genes for flavonoid biosynthesis were up-regulated in the sapwood-heartwood transition zone. This analysis also established the expression patterns of 341 previously unknown genes.     

Application of centrifugal force to the extraction and separation of parasorboside and gerberin from Gerbera hybrida

A method for the separation of parasorboside and gerberin from the ornamental plant Gerbera hybrida (Asteraceae) has been developed. The two closely related glucosides were extracted using an Extrachrom instrument, a prototype multi-functional separation tool equipped with an extraction chamber. The rotation planar extraction procedure was compared with that of a medium pressure solid-liquid extraction system. The resulting extracts were pre-purified using rotation planar chromatography and the results compared with those obtained using medium pressure liquid chromatography with silica gel as the stationary phase and a mobile phase of methanol:ethyl acetate:tetrahydrofuran at selectivity point Ps = 111 with 1% formic acid as modifier. The title compounds were isolated from the purified extracts by TLC and their structures confirmed by 1H- and 13C-NMR spectroscopy.