Lignin-like Compounds Research Articles

A Novel Method for Unraveling the Black Box of Dissolved Organic Matter in Soils by FT-ICR-MS Coupled with Induction-Based Nanospray Ionization

Dissolved organic matter (DOM) in soils plays important roles in carbon and nutrient cycling. However, our knowledge of the molecular composition of DOM is limited by selective loss during pretreatment [solid-phase extraction (SPE)] or small amounts of pore water. We proposed a novel method for characterizing DOM by coupling a Fourier transform ion cyclotron resonance mass spectrometer and a self-built induction-based nanospray ionizer. This approach reduces the ion suppression effect and avoids the need for SPE by substantially reducing the matrix effects. Moreover, highly diverse compound types were detected in Suwannee River natural organic matter. Cwₜ/Nwₜ was close to the actual C/N ratio. Only 0.5 μL of pore water (5 ng of C) was needed (vs ∼10 mL for the conventional method). This method was validated by analyzing forest and agricultural soil samples. Soil DOM was not dominated by lignin-like compounds, as determined using the conventional method. The novel method revealed more peptide/carbohydrate-like compounds, highly condensed aromatics, and more than double the number of CHNO compounds. A more complete understanding of DOM will allow us to elucidate the biogeochemical processes of nutrient and DOM–pollutant complexes in soils as well as to develop comprehensive policies for soil remediation.

Applicability of Recombinant Laccases From the White-Rot Fungus Obba rivulosa for Mediator-Promoted Oxidation of Biorefinery Lignin at Low pH.

Utilization of lignin-rich side streams has been a focus of intensive studies recently. Combining biocatalytic methods with chemical treatments is a promising approach for sustainable modification of lignocellulosic waste streams. Laccases are catalysts in lignin biodegradation with proven applicability in industrial scale. Laccases directly oxidize lignin phenolic components, and their functional range can be expanded using low-molecular-weight compounds as mediators to include non-phenolic lignin structures. In this work, we studied in detail recombinant laccases from the selectively lignin-degrading white-rot fungus Obba rivulosa for their properties and evaluated their potential as industrial biocatalysts for the modification of wood lignin and lignin-like compounds. We screened and optimized various laccase mediator systems (LMSs) using lignin model compounds and applied the optimized reaction conditions to biorefinery-sourced technical lignin. In the presence of both N–OH-type and phenolic mediators, the O. rivulosa laccases were shown to selectively oxidize lignin in acidic reaction conditions, where a cosolvent is needed to enhance lignin solubility. In comparison to catalytic iron(III)–(2,2,6,6-tetramethylpiperidin-1-yl)oxyl (TEMPO) oxidation systems, the syringyl-type lignin units were preferred in mediated biocatalytic oxidation systems.

Organic amendments change soil organic C structure and microbial community but not total organic matter on sub-decadal scales

Organic C has many benefits for soil, but it is depleted by tillage and crop harvest, and especially so for biofuel crops. Accordingly, strategies such as partially retaining stover or planting a cover crop can help ameliorate the negative effect of C removal. We used a long-term field experiment to study the impacts of stover retention and planting a cover crop on soil organic matter (SOM), its extractable components, and the soil microbial community. SOM chemical composition characterization was determined by electrospray ionization (ESI) coupled with Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) in sequential water, methanol (MeOH), and chloroform (CHCl3) extracts. The characteristics of the soil bacterial community were measured by phospholipid fatty acid (PLFA), real-time quantitative PCR, and 16S rRNA gene sequence. The variations in total SOM content, total microbial biomass, and bacterial population were slight among treatments, but SOM chemical compounds, arbuscular mycorrhizal fungi (AMF) biomass, and bacterial structure changed significantly, and especially so in the coupled application of stover retention and cover crop. Specifically, stover retention enriched more lignin-like compounds in soil, whereas cover crop enriched more condensed hydrocarbons, and had more compounds with an aromaticity index (AI) > 0.5. The bacterial community was not altered by the cover crop, but the corn stover retention increased the relative abundances of Myxococcales (Deltaproteobacteria) and decreased that of Actinobacteria. Redundancy analysis (RDA) further revealed that the bacterial community in the stover treatments had a significant positive association with CHCl3-extracted chemical classes, i.e. unsaturated hydrocarbons and lipids, with the coupled application (stover and cover crop), and lignin and proteins with the corn stover only treatment. Taken together, our study shows how different C addition practices influence the molecular composition of SOM and the structure of soil microbial communities.

Use of UV resonance Raman spectroscopy for assessing the brightness stability of ozone TCF bleached pulp

Abstract This study intends to explain the difference in brightness stability between hardwood ECF and TCFz kraft pulps bleached by DEpDD and A(ZEo)(ZEo)(ZP) sequences respectively, using UV Resonance Raman (UVRR) spectroscopy. The brightness stability of the pulps was tested via dry aging experiments where the Post-Color Number (PCN) of the ECF pulp was twice that of the TCF pulp. The aged and non-aged bleached pulps were analyzed with UVRR spectroscopy to identify the cause of the large difference in PCN. The spectra of ECF and TCF bleached pulps presented clear differences in the intensities of the Raman shifts associated to lignin, lignin-like compounds, and degradation products such as muconic acids. To identify more specifically the compounds affecting the PCN, several post-bleaching treatments were applied on the ECF pulp including single stages (E, B, P, Z) or combinations (ZE, ZB, ZP), and their UVRR spectra analyzed. It was found that alkaline-soluble compounds were the main culprits for the difference in PCN values between ECF and TCFz pulps. ZP combination was the most efficient in eliminating residual lignin and other unsaturated components and for the development of brightness and brightness stability.

Vegetation affects photoprotective pigments and copepod distribution in the Himalayan lakes: Implication for climate change adaptation

Lakes in the Himalayas host unique biota and biological communities which are highly sensitive to climate change. High penetration of solar UV radiation in clear shallow lake affects the distribution and abundance of the zooplankton communities. Survival of copepods in such habitats often relies on available photoprotective compounds. We estimated species diversity and distribution patterns of copepods with detectable carotenoids in 7 lakes of the central Himalayas along the altitudinal gradients. To determine the factors influencing the accumulation of high-level photoprotective compounds, we analyzed the physicochemical parameters of water and the concentration of Lignin like Compounds (LLCs), Aromaticity (ARO), Humic Compounds (HCs), Degree of Humification (DoH) and percent Total Organic Matter (TOM) in littoral sediments. In the shallow lakes, copepod abundance and diversity correlate with water transparency. Humic compounds (HCs) derived from ligninaceous plants stimulate the accumulation of photoprotective compounds that allow for the domination of diaptomidae. Copepods receive photoprotective compounds from the humic-bounded sediment substrate. The amount of photoprotectants in the aquatic food chain of the central Himalayas is largely influenced by ligninaceous compounds derived from catchment vegetation. In copepods of shallow clear lakes, the remnant of dead trees in the littoral zones helps to minimize hazards caused by exposure effect and climatic stress. The reduction of vegetative covers in the shorelines may alter the community structure of zooplankton, particularly in the upland ecosystem.

Molecular Characteristics of Dissolved Organic Nitrogen and Its Interaction with Microbial Communities in a Prechlorinated Raw Water Distribution System.

Dissolved organic nitrogen (DON) represents a unique challenge in prechlorinated raw water distribution systems (PRWDSs) because of its contribution to the formation of harmful nitrogen-disinfection byproducts, influence upon biogeochemical processes, and unclear molecular characteristics. Here, Fourier transform ion cyclotron resonance mass spectrometry in combination with high-throughput sequencing was applied to elucidate the molecular changes of DON and biofilm microbial communities in a PRWDS in Yixing, China. Our study revealed that dynamic characteristics of DON are significantly correlated with the biofilm. The accumulation of refractory lignin-like compounds and CnHmOpN1 contributes to the higher recalcitrance molecular characteristics of DON in the effluent associated with Alphaproteobacteria, Planctomycetes, and Bacteroidetes. Additionally, with the help of prechlorination, the biofilm may change the DON characteristics and lead to higher oxygenation, higher m/z, and lower saturation during transportation. Despite the promotion of CnHmOpN1 and CnHmOpN3 at the early stage, we suggest that appropriate concentration of chlorine can add to the front end of raw water distribution pipes. Prechlorination may control the nitrification process and stabilize the rapid growth of diversity and concentration of low molecular weight DON, especially the refractory CnHmOpN1 in the effluent, which may help to improve treatment efficiency of drinking water treatment plants.

Tatarinan T, an α-asarone-derived lignin, attenuates osteoclastogenesis induced by RANKL via the inhibition of NFATc1/c-Fos expression.

We have previously reported that the lignin-like compounds, Tatarinan O (TO) and Tatarinan N (TN), extracted from the roots of Acorus tatarinowii Schott, inhibit receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclastogenesis. In the present study, the potential function of the α-asarone-derived lignins, Tatarinan T (TT) and Tatarinan A (TA), to regulate RANKL-induced osteoclastogenesis was investigated, and it was found that only early treatment with TT may inhibit RANKL-triggered formation of osteoclasts and resorption. The results revealed repressed expression levels of several osteoclast marker genes, including ATPase H+ -transporting V0 subunit d2 (Atp6v0d2), αvβ3 integrin, and osteoclast-associated receptor (OSCAR), following TT treatment during osteoclastogenesis. Moreover, TT reduced the expression levels of the core transcription elements, nuclear factor of activated T-cells cytoplasmic 1 (NFATc1) and c-Fos. However, western blotting analysis showed that TT treatment did not alter nuclear factor-κΒ (NF-κB) activation or mitogen-activated protein kinase (MAPK) or Syk/Btk/phospholipase Cγ2 (PLCγ2) phosphorylation. Taken together, these results suggest the potential of TT in the treatment of diseases of increased bone resorption.

Potential use of green alga Ulva sp. for papermaking

The large amount of cellulose found in Ulva sp. and its low percentage of lignin-like compounds make it an interesting raw material for partially substituting wood pulp to produce pulp and paper. This work shows the suitability of mild chemical treatments for papermaking using residual biomass from this green seaweed, harvested on the beaches, in order to give it added value. A chemical characterization was used to determine ethanol-benzene, hot water, and 1% soda extractives, ash content, holocellulose, α-cellulose, and acid-insoluble material. Cellulose extraction was performed with low proportions of soda and hydrogen peroxide, and it was subjected to a refining step. A design of experiments was used to explain the influence of soda (6%, 8%, and 10%) and hydrogen peroxide (2%, 4%, and 6%) based on oven-dry weight, plus refining (1000 PFI revolutions, 3000 PFI revolutions, and 5000 PFI revolutions). The results showed that to attain good paper strength, it is advisable to operate at maximum alkali charge, minimum peroxide concentration, and refine to a high degree.

Molecular characterization of organic aerosol in the Himalayas: insight from ultra-high-resolution mass spectrometry

Abstract. An increased trend in aerosol concentration has been observed in the Himalayas in recent years, but the understanding of the chemical composition and sources of aerosol remains poorly understood. In this study, molecular chemical composition of water-soluble organic matter (WSOM) from two filter samples collected during two high aerosol loading periods (denoted as P1 and P2) at a high-altitude station (Qomolangma Station, QOMS; 4276 m a.s.l.) in the northern Himalayas was identified using electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR MS). More than 4000 molecular formulas were identified in each filter sample which were classified into two compound groups (CHO and CHON) based on their elemental composition, with both accounting for nearly equal contributions in number (45 %–55 %). The relative abundance weighted mole ratio of O∕Cw for P1 and P2 was 0.43 and 0.39, respectively, and the weighted double bond equivalents (DBEw), an index for the saturation of organic molecules, were 7.12 and 7.87, respectively. Although the O∕Cw mole ratio was comparable for CHO and CHON compounds, the DBEw was significantly higher in CHON compounds than CHO compounds. More than 50 % molecular formulas in the Van Krevelen (VK) diagram (H∕C vs. O∕C) were located in 1–1.5 (H∕C) and 0.2–0.6 (O∕C) regions, suggesting potential lignin-like compounds. The distributions of CHO and CHON compounds in the VK diagram, DBE vs. number of C atoms, and other diagnostic diagrams showed high similarities among each other, suggesting their similar source and/or atmospheric processes. Many formulas formed from biogenic volatile organic compounds (e.g., ozonolysis of α-pinene products) and biomass-burning-emitted compounds (e.g., phenolic compounds) were found in the WSOM, suggesting the important contribution of these two sources in the Himalayas. The high DBE and high fraction of nitrogen-containing aerosol can potentially impact aerosol light absorption in this remote region. Further comprehensive study is needed due to the complexity of organic aerosol and limited molecular number identified in this study.

Redox Fluctuations Control the Coupled Cycling of Iron and Carbon in Tropical Forest Soils.

Oscillating redox conditions are a common feature of humid tropical forest soils, driven by an ample supply and dynamics of reductants, high moisture, microbial oxygen consumption, and finely textured clays that limit diffusion. However, the net result of variable soil redox regimes on iron (Fe) mineral dynamics and associated carbon (C) forms and fluxes is poorly understood in tropical soils. Using a 44-day redox incubation experiment with humid tropical forest soils from Puerto Rico, we examined patterns in Fe and C transformations under four redox regimes: static anoxic, "flux 4-day" (4d oxic, 4d anoxic), "flux 8-day" (8d oxic, 4d anoxic) and static oxic. Prolonged anoxia promoted reductive dissolution of Fe-oxides, and led to an increase in soluble Fe(II) and amorphous Fe oxide pools. Preferential dissolution of the less-crystalline Fe pool was evident immediately following a shift in bulk redox status (oxic to anoxic), and coincided with increased dissolved organic C, presumably due to acidification or direct release of organic matter (OM) from dissolving Fe(III) mineral phases. The average nominal oxidation state of water-soluble C was lowest under persistent anoxic conditions, suggesting that more reduced organic compounds were metabolically unavailable for microbial consumption under reducing conditions. Anoxic soil compounds had high H/C values (and were similar to lignin-like compounds) whereas oxic soil compounds had higher O/C values, akin to tannin- and cellulose-like components. Cumulative respiration derived from native soil organic C was highest in static oxic soils. These results show how Fe minerals and Fe-OM interactions in tropical soils are highly sensitive to variable redox effects. Shifting soil oxygen availability rapidly impacted exchanges between mineral-sorbed and aqueous C pools, increased the dissolved organic C pool under anoxic conditions implying that the periodicity of low-redox events may control the fate of C in wet tropical soils.

Tatarinan N inhibits osteoclast differentiation through attenuating NF-κB, MAPKs and Ca2+-dependent signaling

Osteoclasts are multinucleated cells that originate from hemopoietic stem cells. Targeting over activated osteoclasts is thought to be an effective therapeutic approach to osteoporosis. In a previous study, we reported that Tatarinan O, a lignin-like compound, suppressed RANKL-induced osteoclastogenesis. In this study, we further examined the effects on osteoclast formation of three lignin-like compounds including Tatarinan N (TN), Tatarinan U (TU) and Tatarinan V (TV), all containing a common structure of asarone. We found that only TN suppressed RANKL-induced osteoclast differentiation, bone resorption pit formation and F-acting ring formation. TU and TV did not influence RANKL-induced osteoclastogenesis. We also found that TN dose-dependently inhibited the expression of osteoclastogenesis-associated genes, including TRAP, cathepsin K and MMP-9. Furthermore, we found that TN down-regulated the key transcription factor NFATc1 and c-Fos by preventing the activation of NF-κB and phosphorylation of MAPKs including ERK1/2 and p38 but not JNK. TN attenuated calcineurin expression via suppression of the Btk-PLCγ2 cascade and reduction of intracellular Ca2+, modulating NFATc1 activation. Taking together, our results indicated that TN might have therapeutic potential for osteoporosis.

Distribution of Lignin, Hemicellulose, and Arabinogalactan Protein in Hemp Phloem Fibers.

The distribution of lignin, 8-5' and 8-8' linked lignin substructure, and noncellulosic polysaccharides in hemp (Cannabis sativa L.) phloem fibers were explored based on histochemical and immunological methods. Ultraviolet absorption and potassium permanganate staining were observed mainly in the compound middle lamella (CML) and S1 layers, and rarely in the G-layer of phloem fibers, suggesting that lignin concentration is high at the CML and S1 layers, and very low at the G-layer of hemp fibers. Acriflavine staining, uniform KM1 labeling (8-5' linked lignin substructure), and no KM2 labeling (8-8' linked structure) were observed in the G-layer, suggesting that there is a small amount of lignin-like compound with 8-5' linked structure in the G-layer. In addition, some fiber cells showed a multilayered structure. Uniform arabinogalactan protein (AGP) labeling was observed on the S1 layers and G-layers using JIM14, but little appeared in the CML of hemp fibers, indicating that these layers of the phloem fibers contain AGP. Immunogold labeling of xylan (LM11) and glucomannan (LM21) showed that xylan and glucomannan were mainly present in the S1 layers and the G-layers, respectively. In some phloem fibers, LM21 immunofluorescence labeling showed multilayered structure, suggesting the heterogeneous distribution of glucomannan.

Characterisation of dissolved organic matter using Fourier-transform ion cyclotron resonance mass spectrometry: Type-specific unique signatures and implications for reactivity

This study investigated the chemodiversity and unique signatures for dissolved organic matter (DOM) from different types of water using high-resolution Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS). Eight freshwater hydrophobic DOM (HPO) share 10–17% formulas that were mainly lignin-like compounds. Unique signatures were synthesized: unique to the high-humic HPO (Suwannee River and Blavet River) were condensed aromatic and some aliphatic compounds with H/C > 1.5 and O/C < 0.2, which is considered as decisive of these black river water. Medium-humic isolates (Loire River, Seine River, South Platte River, and Ribou Dam) did not show explicit unique signatures. Nonetheless, enhanced chemodiversity was observed for medium-humic isolates extracted from a variety of indigenous environmental conditions. As an example, South Platte River HPO isolated in winter showed signatures similar to low-humic HPO (Colorado River), i.e., predominantly aliphatic CHO (H/C > 1.0). Effluent HPO was mainly aliphatic molecules with 0.2 < O/C < 0.5 and enriched in S-bearing molecules, and molecules unique to glacial DOM (Pony Lake) incorporated N-bearing compounds that were inferiorly oxidized and were considered as microbial-derived. The weight-averaged double bond equivalent and elemental ratio derived from FTICR-MS were compared with SUVA254 and the results from elemental analysis. This acts as the first study to synthesize unique chemical compositions that distinguish different types of DOM and determine certain reactivity. It is also a significant reference for future studies using similar types of DOM.

Microbial Interactions With Dissolved Organic Matter Drive Carbon Dynamics and Community Succession.

Knowledge of dynamic interactions between natural organic matter (NOM) and microbial communities is critical not only to delineate the routes of NOM degradation/transformation and carbon (C) fluxes, but also to understand microbial community evolution and succession in ecosystems. Yet, these processes in subsurface environments are usually studied independently, and a comprehensive view has been elusive thus far. In this study, we fed sediment-derived dissolved organic matter (DOM) to groundwater microbes and continually analyzed microbial transformation of DOM over a 50-day incubation. To document fine-scale changes in DOM chemistry, we applied high-resolution Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) and soft X-ray absorption spectroscopy (sXAS). We also monitored the trajectory of microbial biomass, community structure and activity over this time period. Together, these analyses provided an unprecedented comprehensive view of interactions between sediment-derived DOM and indigenous subsurface groundwater microbes. Microbial decomposition of labile C in DOM was immediately evident from biomass increase and total organic carbon (TOC) decrease. The change of microbial composition was closely related to DOM turnover: microbial community in early stages of incubation was influenced by relatively labile tannin- and protein-like compounds; while in later stages the community composition evolved to be most correlated with less labile lipid- and lignin-like compounds. These changes in microbial community structure and function, coupled with the contribution of microbial products to DOM pool affected the further transformation of DOM, culminating in stark changes to DOM composition over time. Our study demonstrates a distinct response of microbial communities to biotransformation of DOM, which improves our understanding of coupled interactions between sediment-derived DOM, microbial processes, and community structure in subsurface groundwater.

Molecular Insights into Arctic Soil Organic Matter Degradation under Warming.

Molecular composition of the Arctic soil organic carbon (SOC) and its susceptibility to microbial degradation are uncertain due to heterogeneity and unknown SOC compositions. Using ultrahigh-resolution mass spectrometry, we determined the susceptibility and compositional changes of extractable dissolved organic matter (EDOM) in an anoxic warming incubation experiment (up to 122 days) with a tundra soil from Alaska (United States). EDOM was extracted with 10 mM NH4HCO3 from both the organic- and mineral-layer soils during incubation at both -2 and 8 °C. Based on their O:C and H:C ratios, EDOM molecular formulas were qualitatively grouped into nine biochemical classes of compounds, among which lignin-like compounds dominated both the organic and the mineral soils and were the most stable, whereas amino sugars, peptides, and carbohydrate-like compounds were the most biologically labile. These results corresponded with shifts in EDOM elemental composition in which the ratios of O:C and N:C decreased, while the average C content in EDOM, molecular mass, and aromaticity increased after 122 days of incubation. This research demonstrates that certain EDOM components, such as amino sugars, peptides, and carbohydrate-like compounds, are disproportionately more susceptible to microbial degradation than others in the soil, and these results should be considered in SOC degradation models to improve predictions of Arctic climate feedbacks.

Dissolved organic matter compositions in 0.6–3.4 km deep fracture waters, Kaapvaal Craton, South Africa

The composition of dissolved organic matter (DOM) in extremely deep groundwaters has rarely been investigated. Focusing on deep fracture waters in billion year old rocks in South Africa, we hypothesized the DOM would be predominantly derived from in situ microbial sources, consistent with H2-driven lithoautotrophic ecosystems previously documented in these fluids. We collected groundwater from 0.6 to 3.4 km depths via boreholes in mines, and characterized the DOM by a variety of analytical methods. Dissolved organic carbon concentrations ranged from 0.25 to 4.9 mg C/L. Low molecular weight organic acids (formate, acetate, lactate, propanoate) and amino acids (after hydrolysis) accounted for varying fractions of the total dissolved organic carbon. UV spectrophotometry, scanning fluorimetry, and nuclear magnetic resonance (NMR) spectra indicated lower concentrations of aromatic compounds than are commonly found in surface waters and shallow groundwaters. Molecular weights of aromatic compounds measured by size-exclusion chromatography were dominantly 500–1000 Da. Scanning fluorimetry revealed humic substances in two shallow (578 and 1300 m) samples; deeper samples showed peaks indicating aromatic amino acids. Intensive study of one sample from a previously sampled borehole in Beatrix mine resulted in large NMR peaks for aliphatics and carboxyls with lesser aromatics peaks; negative electrospray ion Fourier transform ion cyclotron resonance-mass spectrometry detected protein-, lipid-, and lignin-like compounds. These results are consistent with DOM in the form of microbial metabolites, and modified cell components (membrane lipids and proteins). These findings support previous geomicrobiological reports of chemolithoautotrophic microbial ecosystems functioning in isolation from photosynthetic primary production.

The fungal ability for biobleaching/biopulping/bioremediation of lignin-like compounds of agro-industrial raw material

Lignin is present in plant cell secondary wall, associated to carbohydrates preventing their efficient hydrolysis, and cellulose pulp manufacture basically consists in breaking down the middle lamella of plant cells, individualizing fibers such as cellulose from the other biopolymers. Different levels of lignocellulose are found in plant residues and they can be decomposed by extracellular fungal lignin modifying enzymes, used as a tool to reduce waste materials in contaminated soils and effluents. In the paper mill industries, for instance, they are a suitable or complementary alternative to the traditional methods of pulping/bleaching, contributing to improve paper strength as well as to reduce the pitch content, the quantity of chemicals and the consume of electrical energy. The aim of this review was to describe the fungal degradation of lignocellulosic like-material, the non-specific enzymatic aspects of the attack of wood and agricultural wastes, the fungal ability for biosorption and bioconversion, and its applications in the pulp/paper industry and bioremediation.

Characterization of selected municipal solid waste components to estimate their biodegradability

Biological treatments of Residual Municipal Solid Waste (RMSW) allow to divert biodegradable materials from landfilling and recover valuable alternative resources. The biodegradability of the waste components needs however to be assessed in order to design the bioprocesses properly. The present study investigated complementary approaches to aerobic and anaerobic biotests for a more rapid evaluation. A representative sample of residual MSW was collected from a Mechanical Biological Treatment (MBT) plant and sorted out into 13 fractions according to the French standard procedure MODECOM™. The different fractions were analyzed for organic matter content, leaching behavior, contents in biochemical constituents (determined by Van Soest's acid detergent fiber method), Biochemical Oxygen Demand (BOD) and Bio-Methane Potential (BMP). Experimental data were statistically treated by Principal Components Analysis (PCA). Cumulative oxygen consumption from BOD tests and cumulative methane production from BMP tests were found to be positively correlated in all waste fractions. No correlation was observed between the results from BOD or BMP bioassays and the contents in cellulose-like, hemicelluloses-like or labile organic compounds. No correlation was observed either with the results from leaching tests (Soluble COD). The contents in lignin-like compounds, evaluated as the non-extracted RES fraction in Van Soest's method, was found however to impact negatively the biodegradability assessed by BOD or BMP tests. Since cellulose, hemicelluloses and lignin are the polymers responsible for the structuration of lignocellulosic complexes, it was concluded that the structural organization of the organic matter in the different waste fractions was more determinant on biodegradability than the respective contents in individual biopolymers.

How Embryophytic is the Biosynthesis of Phenylpropanoids and their Derivatives in Streptophyte Algae?

The origin of land plants from algae is a long-standing question in evolutionary biology. It is becoming increasingly clear that many characters that were once assumed to be 'embryophyte specific' can in fact be found in their closest algal relatives, the streptophyte algae. One such case is the phenylpropanoid pathway. While biochemical data indicate that streptophyte algae harbor lignin-like components, the phenylpropanoid core pathway, which serves as the backbone of lignin biosynthesis, has been proposed to have arisen at the base of the land plants. Here we revisit this hypothesis using a wealth of new sequence data from streptophyte algae. Tracing the biochemical pathway towards lignin biogenesis, we show that most of the genes required for phenylpropanoid synthesis and the precursors for lignin production were already present in streptophyte algae. Nevertheless, phylogenetic analyses and protein structure predictions of one of the key enzyme classes in lignin production, cinnamyl alcohol dehydrogenase (CAD), suggest that CADs of streptophyte algae are more similar to sinapyl alcohol dehydrogenases (SADs). This suggests that the end-products of the pathway leading to lignin biosynthesis in streptophyte algae may facilitate the production of lignin-like compounds and defense molecules. We hypothesize that streptophyte algae already possessed the genetic toolkit from which the capacity to produce lignin later evolved in vascular plants.

Isolation and characterization of Rhizobium sp. strain YS-1r that degrades lignin in plant biomass.

The aim of this work was to isolate novel lignin-degrading organisms. Several pure cultures of bacteria that degrade lignin were isolated from bacterial consortia developed from decaying biomass. Among the isolates, Rhizobium sp. strain YS-1r (closest relative of Rhizobium petrolearium strain SL-1) was explored for its lignin-degrading ability. Microcosm studies showed that strain YS-1r was able to degrade a variety of lignin monomers, dimers and also native lignin in switchgrass and alfalfa. The isolate demonstrated lignin peroxidase (LiP) activity when grown on alkali lignin, p-anisoin, switchgrass or alfalfa, and only negligible activity was measured in glucose-grown cells suggesting inducible nature of the LiP activity. Analysis of the strain YS-1r genome revealed the presence of a variety of genes that code for various lignin-oxidizing, H2 O2 -producing as well as polysaccharide-hydrolysing enzymes. This study shows both the genomic and physiological capability of bacteria in the genus Rhizobium to metabolize lignin and lignin-like compounds. This is the first detailed report on the lignocellulose-degrading ability of a Rhizobium species and thus this study expands the role of alpha-proteobacteria in the degradation of lignin. The organism's ability to degrade lignin is significant since Rhizobia are widespread in soil, water and plant rhizospheres and some fix atmospheric nitrogen and also have the ability to degrade aromatic hydrocarbons.