Eucalyptus globulus wood fractionation by autohydrolysis and organosolv delignification

This work provides an assessment on the fractionation of Eucalyptus globulus wood by sequential stages of autohydrolysis (to cause the solubilization of hemicelluloses) and organosolv pulping (to dissolve lignin, leaving solids enriched in cellulose). With this approach, valuable products (hemicellulose-derived saccharides, sulphur-free lignin fragments and cellulosic substrates with low contents of residual hemicelluloses) are obtained in separate streams, according to the biomass refinery approach. Autohydrolysis was carried out under optimized operational conditions, and organosolv pulping was performed using uncatalyzed ethanol-water solutions. The effects of the most influential operational variables (autohydrolysis severity, delignification temperature and ethanol concentration in the organosolv stage) on solid yield, solid composition, cellulose susceptibility and recovery of the various fractions was assessed using statistical methods, which enabled the identification of the most favourable operational conditions.     

Integral valorization of Leucaena diversifolia by hydrothermal and pulp processing

Wood from the leguminous tree, Leucaena diversifolia, was subjected to hydrothermal treatment (autohydrolysis) at 160-180 °C for 0-30 min followed by ethanol-soda-anthraquinone delignification. The liquid phase contained 18.65 g of sugars per liter, and the solid phase had a gross heating value of 19.083 MJ/kg, but could also be used as a source of cellulose pulp for the production of paper with tear, burst, and tensile indexes of 2.4 N m²/kg, 2.6 MPa m²/kg and 40.7 kN m/kg, respectively. L. diversifolia lends itself readily to valorization for energy production, and also to integral, fractional exploitation by autohydrolysis and ethanol-soda-anthraquinone delignification, which can additionally bring environmental benefits to cropping zones.     

RAPD characterisation of two neotropical hybrid legumes

An investigation of randomly amplified polymorphic DNA (RAPD) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) marker distribution was made for two well-characterised hybrids and their parents,Leucaena leucocephala andL. esculenta andParkinsonia aculeata andCercidium praecox. Three chloroplast DNA (cpDNA) markers identified the maternal parent of eachL. leucocephala ×L. esculenta hybrid. Fifteen species-diagnostic RAPD markers (invariant in one taxon and absent from the other) were always present in theLeucaena hybrid and assumed to be of nuclear origin, whilst three RAPD markers showed expression patterns identical to the cpDNA markers and were assumed to be of organellar origin. No RAPD or PCR-RFLP taxon-diagnostic markers were discovered for eitherP. aculeata orC. praecox. However, 21 RAPD markers were species-specific (polymorphic within one taxon but absent from the other) and Southern analysis indicated that none of the markers were of organellar origin. Only 67% additivity of markers specific toP. aculeata andC. praecox was demonstrated in the hybrids between these two species, whilst inLeucaena 97% additivity was demonstrated. Differences between the two hybridising situations were related to the behaviour of the molecular markers and the biology of the species.     

Improvement of pressure chamber measurements of two legumes by constriction of stems

Summary A valid determination of the balancing pressure end-point was not possible withStylosanthes hamata (L.) Taub. cv. Verano andS. scabra Vog. using a sealing stopper of the Waring and Cleary type. An exudate, apparently from the pith, occurred at low pressures even in stressed plants. The problem did not occur in a chamber fitted with a compression gland. A modification of the Waring and Cleary chamber which caused greater stem constriction proved successful.     

Polymer properties of softwood organosolv lignins produced in two different reactor systems

Lignin, the second most abundant biopolymer on earth and with a predominantly aromatic structure, has the potential to be a raw material for valuable chemicals and other bio-based chemicals. In industry, lignin is underutilized by being used mostly as a fuel for producing thermal energy. Valorization of lignin requires knowledge of the structure and different linkages in the isolated lignin, making the study of structure of lignin important. In this article, lignin samples isolated from two types of reactors (autoclave reactor and displacement reactor) were analyzed by FT-IR, size exclusion chromatography, thermogravimetric analysis (TGA), and Py-GC-MS. The average molecular mass of the organosolv lignins isolated from the autoclave reactor decreased at higher severities, and FT-IR showed an increase in free phenolic content with increasing severity. Except for molecular mass and molecular mass dispersity, there were only minor differences between lignins isolated from the autoclave reactor and lignins isolated from the displacement reactor. Carbohydrate analysis, Py-GC–MS and TGA showed that the lignin isolated using either of the reactor systems is of high purity, suggesting that organosolv lignin is a good candidate for valorization.     

Nitrogen-15 labeling effectiveness of two tropical legumes

Nitrogen-15 foliar applications for the production of field-labeled plant tissues may achieve more effective labeling of plant shoot and root tissues and minimize directly labeling the soil N fraction as occurs when¹⁵ N is soil applied. Consequently, foliar-labeled plant tissues should be better suited for subsequent ¹⁵N mineralization studies. A field experiment was conducted to determine the effectiveness of ¹⁵N-labeling and the accumulation of ¹⁵N in various plant parts of two tropical legumes. Desmodium ovalifolium Guillemin and Perrottet and Pueraria phaseoloides (Roxb.) Benth., grown in 0.5 m² microplots, were labeled with foliar-applied urea containing 99 atom% ¹⁵N. Plants in each microplot received a total of 0.1698 g ¹⁵N that was applied all at once or split equally into two, three or four applications. Legume shoots and roots and soil were destructively harvested and analyzed for total ¹⁵N content. Averaged over both legumes and foliar application rates, total plant (shoots, flowers, leaf litter, and roots) recovery was approximately 79% of the ¹⁵N applied. The soil contained 3% of the ¹⁵N applied, of which 2.5 and 0.5% were in the inorganic and organic fractions, respectively. Nitrogen-15 recovery in shoots (76%) was sixty-five fold greater than in roots (1%) and about nineteen fold greater than the sum of roots and soil (4.1%), a much greater percent recovery than observed in other foliar labeling studies. Averaged over all four foliar split-application rates, ¹⁵N recovery by Desmodium shoots was greater than Pueraria. Results demonstrate that ¹⁵N foliar application to legumes is an effective method for labeling, resulting in atom% excess ¹⁵N levels and ¹⁵N recoveries comparable to those reported with the more traditional soil-labeling approach. Another advantage of this method is a nondestructive, in situ labeling method that permits separation of shoot and root residual N contribution to subsequent crops in N tracer studies.     

Production of oligosaccharides by autohydrolysis of brewery's spent grain

Brewery's spent grain was treated with water in a process oriented towards the production of xylo-oligosaccharides (XOS). A wide range of temperatures and reaction times were tested and the effects of these operational variables on hemicellulose solubilization and reaction products were investigated. The maximal XOS yield (61% of the feedstock xylan) was obtained at 190 degrees C after 5 min of reaction. Several oligosaccharide mixtures with different molecular weight distributions were obtained depending on temperature and reaction time. Longer reaction times led to decreased oligosaccharide production and enhanced concentrations of monosaccharides, sugar decomposition products and acetic acid. With reaction times leading to the maximal yields of XOS, little decomposition into organic acids and aldehydes was found at all the temperatures assayed. From the composition of processed solids, it was calculated that 63-77% of the initial xylan was selectively solubilized in autohydrolysis treatments.     

Photosynthetic responses of two heliotropic legumes from contrasting habitats

Abstract Photosynthetic responses to light, temperature and leaf-to-air water vapour concentration deficit for Strophostyles helvola (L.) Ell. in an open beach site, and Amphicarpa bracteata (L.) Ell. in two deciduous forest sites were quantified. Photosaturated rates of net CO₂ assimilation were 52.1 ± 4.6, 11.0 ± 1.6 and 4.1 ± 0.3 μmol m^?2s^?2 for plants in beach, roadside and closed canopy sites, respectively. In terms of photosynthesis, plants in the beach site were more tolerant of higher leaf temperatures and water vapour concentration deficits than were plants in forested sites. Heliotropic leaf movements in the beach site reduced calculated total daily transpiration by 2%, increased total daily carbon gain by 8% and reduced the transpiration ratio by 9% relative to an horizontal leaf. During long-term sunflecks in forested sites, heliotropism reduced transpiration by 15%, increased carbon gain by 71% and reduced the transpiration ratio by 50% relative to an horizontal leaf. We hypothesize that heliotropic leaf movements in mesic, high-light, low-nitrogen habitats may increase carbon return on nitrogen investment in photosynthesis, while heliotropism in canopy gaps may represent a morphological mechanism to reduce damage to low-light acclimated photosynthetic systems during long-term sunflecks.     

Nodulation of legumes by rhizobium

Abstract. The formation of nitrogen-fixing nodules on leguminous plants is the culmination of an integrated development involving many plant and bacterial genes. This review focuses on the structure, function and regulation of the bacterial genes involved in the nodulation process. We attempt to interpret recent observations on these genes in the context of signal exchanges which occur between the macro-and micro-symbionts.     

Extraction of sulfated polysaccharides by autohydrolysis of brown seaweed Fucus vesiculosus

The extraction of sulfated polysaccharides (fucoidan) by autohydrolysis (AH) of brown seaweed Fucus vesiculosus was studied. Experimental assays were performed under different conditions of temperature (160 to 200°C) and reaction time (10 to 30 min) according to a 2² central composite design, and the conditions able to maximize the fucoidan yield were selected. The alga degradation and the total sugar yield in the liquor after AH were also determined to each experimental condition. The highest fucoidan yield (～16.5% w/w) was obtained when the AH process was performed at 180°C for 20 min. This product was characterized by high-performance liquid chromatography, infrared analysis spectroscopy, and thermal gravimetric analyses, which verified the presence of fucose and galactose as main components (70:30% mol ratio, in average) and an SO₃ content higher than 20%. AH process under optimum reaction conditions was an effective method to recover fucoidan from F. vesiculosus. The use of this technology brings also important advantages from economical and environmental viewpoints since it does not require the use of chemical solvent and generates less waste when compared to conventional extraction procedures.     

Short-term effects of infestation by two aphid species on plant growth and shoot respiration of three legumes

The short-term effects of infestation by cowpea aphids ( Aphis craccivora Koch) and pea aphids [ Acyrthosiphon pisum (Harris)], both Homoptera: Aphididae, on plant growth and respiration of excised, intact shoots of cowpea [ Vigna unguiculata (L.) Walp. cv. Caloona], broad bean ( Vicia faba L. cv. Aquadulce) and garden pea ( Pisum sativum L. cv. Victory Freezer) seedlings were investigated, but not all plant-aphid combinations were utilized. Root, shoot and plant dry weights were significantly reduced within 10 days in the infested plants. Rates of total shoot respiration were significantly greater in infested plants within 10 days, and the increase was not due to increased alternative pathway activity but, rather, to increased cytochrome pathway activity. It is suggested that the aphid-induced increase in shoot respiration may be due to increased rates of photosynthesis, to substances injected into the phloem by the aphids and/or delayed senescence. These data indicate that aphid-infested shoots had a decreased carbon use efficiency.     

Biogas production and valorization by means of a two-step biological process

The scope of this research work was to investigate biogas production and purification by a two-step bench-scale biological system, consisting of fed-batch pulse-feeding anaerobic digestion of mixed sludge, followed by methane enrichment of biogas by the use of the cyanobacterium Arthrospira platensis. The composition of biogas was nearly constant, and methane and carbon dioxide percentages ranged between 70.5–76.0% and 13.2–19.5%, respectively. Biogas yield reached a maximum value (about 0.4 m³_biogas/kgCOD_i) at 50 days-retention time and then gradually decreased with a decrease in the retention time. Biogas CO₂ was then used as a carbon source for A. platensis cultivation either under batch or fed-batch conditions. The mean cell productivity of fed-batch cultivation was about 15% higher than that observed during the last batch phase (0.035 ± 0.006 g_DM/L/d), likely due to the occurrence of some shading effect under batch growth conditions. The data of carbon dioxide removal from biogas revealed the existence of a linear relationship between the rates of A. platensis growth and carbon dioxide removal from biogas and allowed calculating carbon utilization efficiency for biomass production of almost 95%.     

木质素的微生物解聚与高值转化

木质纤维素是地球上最丰富的可再生资源。我国每年产生约9亿吨农业秸秆,因得不到有效利用,不仅造成资源浪费,也产生了诸多严峻的环境问题。缺少木质素的高效降解和资源化利用技术是限制木质纤维素产业化的主要瓶颈之一。虽然木质素的降解与转化多年来一直都受到关注,但是由于木质素结构的复杂性及异质性,使其高效利用受限。近年来,微生物具有的“生物漏斗”式转化特性为木质素的高值转化和利用提供了新方向。本文就生物质利用研究以来,微生物在木质素解聚与转化方面的研究历程与最新进展进行了简要的回顾与总结,并初步讨论了目前木质素高值转化面临的机遇与挑战。     

两种豆科植物及各器官对不同形态氮的吸收、分配研究

豆科植物在氮素缺乏的荒漠生态系统中大量存在，是该生态系统提供有效氮的中心，也是这一区域重要的先锋物种。该文选择古尔班通古特沙漠广泛分布的弯花黄芪(Astragalus flexus)和镰荚黄芪(Astragalus arpilobus)作为研究对象，分别在0～5、5～15 cm土层添加3种不同形态氮(¹⁵N-NH₄⁺、¹⁵N-NO₃^-、¹⁵N-glycine),研究两种植物及各器官对不同形态氮素的吸收、分配策略。结果表明：(1)在不同土层中，两种植物均偏好吸收硝态氮，并且弯花黄芪、镰荚黄芪对硝态氮的最高吸收速率均为3.26、2.59μg·g^-1·h^-1。(2)在不同土层中，植物各器官间均对不同氮源吸收及分配有显著性差异(P<0.05),弯花黄芪根的¹⁵N吸收量均大于镰荚黄芪的，3种不同形态氮主要分配于叶。(3)在不同土层中，不同氮源对两种植物的贡献率均为^...     

Enhanced enzymatic hydrolysis of lignocellulosic biomass pretreated by low-pressure steam autohydrolysis

Summary The use of low-pressure steam autohydrolysis in the pretreatment of corn stover and hybrid poplar has been assessed. In terms of yield of prehydrolyzed solids, minimal by-product formation and extent of subsequent enzymatic saccharification, the results of low-pressure steam pretreatment were found to be as good as or better than those reported for more severe pretreatment processes. Almost complete saccharification of the cellulose in the prehydrolyzed biomass solids was obtained within 24h with a commercial cellulase preparation — Celluclast. The presence of grinding elements (glass beads) during the enzymatic hydrolysis was found to increase the extent of saccharification by 40% to 50% over controls without any grinding elements.     

Competition amongst rhizobial strains for the colonization and nodulation of two tropical legumes

Summary Paired rhizobial strains supplied in several proportions were used to study inter-strain competition in association with Macroptilium atropurpureum (DC) URB (siratro) and Stylosanthes guianensis (Aubl.) Swartz (Stylo, line I R I 1022). Plants raised from surface sterilized seed, were grown on agar in large cotton-wool plugged tubes, and populations and inter-strain ratios determined in the inoculum and on the root at several times after inoculation. The nodules were mapped in order of appearance and the strains they contained identified at harvest.Related substrains and strains of similar growth habit competed more with each other in the colonization of the root surface than did a fast-growing strain in association with a typical slow grower. Capacity amongst slow-growing strains to dominate a paired competitor in the colonization of the root was a strain characteristic and was not affected by host. It was unrelated to effectiveness in the rhizobium-host association.In 5 of the 7 cases nodulation success could be related quantitatively to root-surface representation and a competitive index calculated; in the remainder one of each pair overwhelmed the other over a wide range of inoculum ratios. It was not possible to relate competitive nodulating success to any single feature of the host: rhizobium symbiosis. In the two most striking cases relationship between competitiveness and N₂-fixing effectiveness was reversed; in others competitiveness difference was as great between equally effective as between strains of differing effectiveness. In the case of Stylo there was a marked dominance of an ineffective over an effective competitor, which might be attributed to greater compatibility, as indicated by faster nodulaton by the ineffective strain. This last result argues against use of mixed inocula including any strain ineffective on any of the hosts for which the inoculum is recommended.Work completed as part requirement for the degree of M. Sc., University of New South Wales;     

Size-number trade-offs and pollen production by Papilionaceous legumes

The intra- and interspecific relations between the size and number of pollen grains were examined for 21 species of papilionaceous legumes to test for the trade-off expected from the subdivision of limited resources. We observed a tradeoff between pollen grain diameter and the number of grains produced per flower within 17 of the 21 species examined. For 12 of these species, the observed partial regression coefficient for In (pollen grain diameter) equaled -3, as expected. The remaining five species exhibited more negative partial regression coefficients than expected. Relations between pollen grain size and number that are more negative than expected may result from a decelerating relation between pollen grain size and the resource investment per grain. Flower production significantly influenced pollen production for only eight species so that pollen production seems to be determined primarily on a per flower basis. Interspecifically, pollen size also varied inversely with pollen number for the 21 species even after the effect of phylogenetic relatedness was removed. Pollen size varies relatively little compared to the number of pollen grains produced per flower and therefore may be the primary target of natural selection. The presence of a trade-off in pollen production probably constrains the evolutionary options by which flowering plants can maximize male success.     

Production of xylose-containing fermentation media by enzymatic post-hydrolysis of oligomers produced by corn cob autohydrolysis

Xylooligomer solutions from autohydrolysis of corn cobs were subjected to an enzymatic post-hydrolysis using commercial enzymes with xylanolytic activity. The effect of temperature and pH on the conversion of xylooligomers into xylose was assessed at low enzyme to substrate ratio. Further experiments to evaluate the influence of enzyme loading were carried out. Balanced mixtures of selected formulations were also used. The xylose solutions obtained by coupling autohydrolysis and enzymatic post-hydrolysis stages contained up to 24 g xylose/l, were free of sugar-dehydration products and, by selecting the enzyme dosage and activities, the acetic acid concentration could be reduced, thus improving their potential fermentability. Regardless of the endo- and exo-activity loadings, the maximum conversion achieved either with single or with mixed commercial formulations, was 80% of the theoretical. This fact suggests the existence of a remaining fraction of substituted xylooligomers accounting for 20% of the initial xylan. A close relationship between deacetylation and xylose generation was also observed.     

Microbial valorization of underutilized and nonconventional waste streams

The growing burden of waste disposal coupled with natural resource scarcity has renewed interest in the remediation, valorization, and/or repurposing of waste. Traditional approaches such as composting, anaerobic digestion, use in fertilizers or animal feed, or incineration for energy production extract very little value out of these waste streams. In contrast, waste valorization into fuels and other biochemicals via microbial fermentation is an area of growing interest. In this review, we discuss microbial valorization of nonconventional, aqueous waste streams such as food processing effluents, wastewater streams, and other industrial wastes. We categorize these waste streams as carbohydrate-rich food wastes, lipid-rich wastes, and other industrial wastes. Recent advances in microbial valorization of these nonconventional waste streams are highlighted, along with a discussion of the specific challenges and opportunities associated with impurities, nitrogen content, toxicity, and low productivity.