Effect of cellulose/hemicellulose and lignin on the bioavailability of toluene sorbed to waste paper

Paper constitutes about 38% of municipal solid waste, much of which is disposed of in landfills. Sorption to such lignocellulosic materials may limit the bioavailability of organic contaminants in landfills. The objective of this study was to identify the effect of individual biopolymers in paper on toluene sorption and bioavailability by subjecting fresh and anaerobically degraded office paper and newsprint to enzymatic hydrolysis and acid hydrolysis. Enzymatic degradation of cellulose and hemicellulose had no effect on toluene bioavailability. In contrast, acid-insoluble lignin controlled toluene sorption and bioavailability for both fresh and degraded newsprint. Acid-insoluble lignin could explain only 54% of the toluene sorption capacity of degraded office paper however, suggesting that crude protein and/or lipophilic organic matter were also important sorbent phases. Toluene sorbed to degraded office paper was also less bioavailable than toluene sorbed to an equivalent mass of lignin extracted from this sorbent. The latter result suggests that a fraction of toluene sorbed to degraded office paper may have been sequestered by lipophilic organic matter. The sorption and bioavailability data indicate that the preferential decomposition of cellulose and hemicellulose relative to lignin in landfills should not decrease the overall toluene sorption capacity of paperwaste or increase the bioavailability of sorbed toluene.     

Molecular characterization of biodegradable dissolved organic matter using bioreactors and [12C/13C] tetramethylammonium hydroxide thermochemolysis GC-MS

Little is known about the molecular composition of the biodegradable fraction of dissolved organic matter (BDOM) in stream ecosystems. We combined plug-flow biofilm reactors, tetramethylammonium hydroxide (TMAH) thermochemolysis GC-MS, and 13C-labeled TMAH thermochemolysis GC-MS to study the molecular composition of BDOM from two stream ecosystems. TMAH products derived from fatty acids, lignin, and other aromatic molecules were quantified using an internal standard approach. We applied the 13C-TMAH thermochemolysis procedure to differentiate between compounds in dissolved organic matter (DOM) that had natural methoxyl groups from those that acquired methoxyl groups during the TMAH reaction. In Rio Tempisquito, a stream draining a tropical evergreen forest, and White Clay Creek, a stream draining a temperate deciduous woodlands, carbohydrates, fatty acids, and lignin contributed to the DOM and BDOM molecular composition. We observed 97 different peaks in the chromatograms of streamwater, with 57% of the peaks common to both streams. The DOM and BDOM pools from each site also contained a unique suite of compounds. Our combined use of TMAH and 13C-TMAH thermochemolysis revealed that heterotrophic bacteria can selectively degrade and demethylate different types of compounds in the lignin residues of DOM. This demonstration of bacterial demethylation of lignin, an abundant and refractory plant molecule, has potential implications for global carbon cycling.     

嗜碱性木素降解菌降解能力的初步研究

通过比较不同嗜碱细菌对麦草中木素、纤维素和半纤维素的降解率, 并比较不同菌株的产酶及酶活情况, 筛选出了木素降解能力较强, 而纤维素和半纤维素降解能力相对较弱的嗜碱性木素降解菌。在ｐＨ≈１０－４的条件下培养８ｄ 后, 菌株６ 降解了３２－３７％ 的麦草木素, 而纤维素和半纤维素分别降解了２１－４８％ 和２２－６９％ 。这与其产酶情况基本一致, 该菌株的酶活最高, 分别为ＭｎＰ２７１－３０Ｕ／Ｌ和ＬｉＰ４１－９４Ｕ／Ｌ。     

Phenanthrene sorption by aliphatic-rich natural organic matter

Contaminant sorption, an important process that may limit bioavailability, hinder remediation, encourage environmental persistence, and control mobility in the environment, has been the focus of numerous studies. Despite these efforts, the fundamental understanding of sorptive processes in soil and sedimentary environments has not been resolved. For instance, many have suggested that contaminants, such as polycyclic aromatic hydrocarbons (PAHs), solely interact with aromatic domains of organic matter. Until now, studies have neglected the aliphatic components that are known to be a recalcitrant and significant part of soil and sedimentary organic matter (SOM). In this investigation, the sorption of phenanthrene to several aliphatic-rich SOM samples was measured. The samples included the following: SOM precursors (algae, degraded algae, cellulose, collagen, cuticle, and lignin), two kerogen samples, and a highly aromatic humic acid. All samples were characterized by cross polarization magic angle spinning carbon-13 (CPMAS 13C) NMR and carbon, hydrogen, and nitrogen analysis. Batch experiments demonstrated that the highest organic carbon normalized sorption coefficients (Koc values) were obtained with the Pula kerogen sample (log Koc = 4.88) that only contains 6.5% aromatic carbon. Other aliphatic-rich samples, namely the Green River kerogen, degraded algae, and collagen samples produced comparable log Koc values (4.64, 4.66, and 4.72, respectively) to that of the highly aromatic humic acid (log Koc = 4.67). Phenanthrene uptake was the least for cellulose and lignin, two major soil components. A comparison of phenanthrene Koc values and paraffinic carbon content revealed a positive correlation (Koc = 798 +/- 96.1 * paraffinic carbon (%), r2 = 0.56) and indicates that amorphous polymethylene carbon is an important consideration in phenanthrene sorption. This study establishes that aliphatic SOM domains have a strong affinity for phenanthrene and likely, other PAHs. Therefore, aliphatic structures, that are an important component of SOM, require more attention in the examination of sorption processes in terrestrial and sedimentary environments.     

Advanced solid-state NMR characterization of marine dissolved organic matter isolated using the coupled reverse osmosis/electrodialysis method

Advanced (13)C solid-state techniques were employed to investigate the major structural characteristics of two surface-seawater dissolved organic matter (DOM) samples isolated using the novel coupled reverse osmosis/electrodialysis method. The NMR techniques included quantitative (13)C direct polarization/magic angle spinning (DP/MAS) and DP/MAS with recoupled dipolar dephasing, (13)C cross-polarization/total sideband suppression (CP/TOSS), (13)C chemical shift anisotropy filter, CH, CH(2), and CH(n) selection, two-dimensional (1)H-(13)C heteronuclear correlation NMR (2D HETCOR), 2D HETCOR combined with dipolar dephasing, and (15)N cross-polarization/magic angle spinning (CP/MAS). The two samples (Coastal and Marine DOM) were collected at the mouth of the Ogeechee River and in the Gulf Stream, respectively. The NMR results indicated that they were structurally distinct. Coastal DOM contained significantly more aromatic and carbonyl carbons whereas Marine DOM was markedly enriched in alkoxy carbon (e.g., carbohydrate-like moieties). Both samples contained significant amide N, but Coastal DOM had nitrogen bonded to aromatic carbons. Our dipolar-dephased spectra indicated that a large fraction of alkoxy carbons were not protonated. For Coastal DOM, our NMR results were consistent with the presence of the major structural units of (1) carbohydrate-like moieties, (2) lignin residues, (3) peptides or amino sugars, and (4) COO-bonded alkyls. For Marine DOM, they were (1) carbohydrate-like moieties, (2) peptides or amino sugars, and (3) COO-bonded alkyls. In addition, both samples contained significant amounts of nonpolar alkyl groups. The potential sources of the major structural units of DOM were discussed in detail. Nonprotonated O-alkyl carbon content was proposed as a possible index of humification.     

白腐菌对稻草秸秆中木质纤维素降解规律的研究

通过对傅里叶变换红外光谱(FTIR)谱图的分析和木质纤维素组分含量变化的测定,研究了3株白腐菌在50天培养期内降解稻草中木质纤维素的规律。结果表明,3株白腐菌对稻草秸秆中木质纤维素的降解具有一定的顺序和选择性,先降解半纤维素和木质素,再同时降解半纤维素、纤维素和木质素。从降解比例来看,白腐菌对半纤维素和木质素具有很好的降解优势,对半纤维素有较好的降解选择性。     

On the nature of the Ca binding to the hull of nixtamalized corn grains

Corn hull is composed of cellulose, hemicelluloses and a minor fraction of lignin. Hemicelluloses are acidic polysaccharides which could explain the relatively large ability of the corn hull to retain Ca during the cooking of corn in a saturated solution of Ca(OH)₂. In order to shed light in this sense the Ca binding to the hull of alkaline cooked corn grains was studied using EDS, pH titration, IR and ¹³C CP/MAS NMR techniques. For short cooking times, prior to hull degradation, Ca is retained in the corn grain according to the following order: hull>germ>endosperm. The acidic groups of hemicelluloses (mainly uronic acids) are the responsible for Ca retention in the corn hull. The amount of Ca retained by the hull approximately is 4 mg/g. On the alkaline cooking the hull is progressively degraded and loses its ability to retain Ca because the hemicelluloses fraction passes to the cooking liquor.     

Sorption of aromatic organic contaminants by biopolymers: effects of pH, copper (II) complexation, and cellulose coating

Sorption of hydrophobic organic compounds (HOCs) (i.e., pyrene, phenanthrene, naphthalene, and 1-naphthol) by original and coated biopolymers was examined. Lignin yielded nonlinear isotherms due to its glassy character. Except for pyrene, cellulose showed linear isotherms for other compounds, indicating a partitioning dominant mechanism. Sorption of 1-naphthol by lignin decreased with increasing pH, attributed to both the increased pi e theta-pi e theta repulsion and weakened hydrogen bonds, while the affinity reduction of cellulose for 1-naphthol with increasing pH resulted from only the decrease in H-bonding due to its absence of benzene ring. Complexation of lignin with Cu2+ increased the sorption affinity for phenanthrene (2.6 times) and slightly enhanced its isotherm nonlinearity. For 1-naphthol, lignin-Cu2+ complex had a much higher sorption capacity (7 times)than the original lignin, accompanied bythe increased isotherm nonlinearity. Cellulose-coated lignin showed increased sorption affinity and more pronounced nonlinearity for 1-naphthol than the lignin-Cu2+ complex. In comparison, cellulose coating exhibited little effect on sorption affinity for phenanthrene relative to the lignin-Cu2+ complex. Isotherm nonlinearity of coated lignins increased with increasing cellulose coating, indicating more condensed domains produced, supported by an increase (from 68.9 degrees C for the original lignin to 82.4 degrees C for the highest cellulose coating level) in glass transition temperature (Tg). Results of this study highlightthe importance of structure, polarity, surface O-containing functional groups, and surface charges of biopolymers in controlling HOC sorption.     

Fungal pretreatment of wheat straw: Effects on the biodegradability of cell walls,structural polysaccharides,lignin and phenolic acids by rumen microorganisms

Disappearance of cell wall components of untreated straw and straw treated with the ligninolytic white-rot fungi Phanerochaete chrysosporium, Dichomitus squalens and Cyathus stercoreus were determined during the course of rumen digestion of samples in nylon bags. The first fungus degraded hemicelluloses and cellulose non-selectively, adversely affecting the digestion rate of crude cell walls. Dichomitus squalens and C. stercoreus preferentially degraded hemicelluloses and lignin, affording cell wall degradation rates 1.5 times higher than in native straw. Furthermore, the extent of cell wall digestion was also significantly enhanced. Both strains improved the extent of cellulose digestion, whereas the potentially degradable xylan fraction remained unchanged. Polysaccharide digestion rates were influenced in different ways depending on the strain tested: straw degraded by C. stercoreus showed an increase in cellulose digestion rate by 50%, whereas residual arabinose units were slowly degraded. Xylan was degraded 1.8 times faster in straw decayed' by D. squalens, while cellulose digestion remained unchanged. Phanerochaete chrysosporium depressed both xylan and cellulose digestion rates. Fungal-treated lignins were solubilised in the rumen faster than in untreated straw, whereas only treatment by C. stercoreus resulted in higher lignin losses. Esterified phenolic acids were extensively degraded by all three fungi. Residual ferulic and p-coumaric acids accumulated during rumen digestion, although only the former decreased in the original straw.     

Absence of mobile carbohydrate domains in dry humic substances proven by NMR, and implications for organic-contaminant sorption models

The mobility and domain structure of various standard humic substances have been investigated by 1H and 1H-13C solid-state nuclear magnetic resonance (NMR) experiments. In four dry humic acids, a fulvic acid, a natural organic matter sample, and a whole peat sample, segments that undergo fast, large-amplitude motions account for <9% of the sample. This disproves a previous suggestion, based on 1H NMR data, that flexible domains, presumably carbohydrates, make up >40% of various humic acids; these putative mobile domains were also linked to dual-mode sorption. In particular, neither the polar alkyl (carbohydrate) nor the aromatic components show any fast, large-amplitude mobility. A small fraction of mobile nonpolar alkyl segments identified by us before is the only component undergoing large-amplitude motions, apart from absorbed water that we observe in humic acids exposed to ambient air. 1H-13C wide-line separation NMR shows that, contrary to previous suggestions, the dipolar couplings in the aromatic regions are smaller than in the polar alkyl segments, most likely due to differences in local 1H densities. Series of 1H-13C heteronuclear correlation experiments with 1H spin diffusion reveal close proximity of aromatic and polar alkyl segments in several humic acids, precluding carbohydrate domains on a scale of > 1-nm diameter. In the standard peat humic acid, nonpolar aromatic segments also do not form sorption domains of significant size, while nonpolar aliphatic domains, which we had previously shown to correlate with sorption capacity, have been confirmed.     

New structural information on a humic acid from two-dimensional 1H-13C correlation solid-state nuclear magnetic resonance

New information on the chemical structure of a peat humic acid has been obtained using a series of two-dimensional 1H-13C heteronuclear correlation solid-state NMR (HETCOR) experiments with different contact times and with spectral editing by dipolar dephasing and 13C transverse relaxation filtering. Carbon-bonded methyl groups (C-CH3) are found to be near both aliphatic and O-alkyl but not aromatic groups. The spectra prove that most OCH3 groups are connected directly with the aromatic rings, as is typical in lignin. As a result, about one-third of the aromatic C-O groups is not phenolic C-OH but C-OCH3. Both protonated and unprotonated anomeric O-C-O carbons are identified in the one- and two-dimensional spectra. COO groups are found predominantly in OCHn-COO environments, but some are also bonded to aromatic rings and aliphatic groups. All models of humic acids in the literature lack at least some of the features observed here. Compositional heterogeneity was studied by introducing 1H spin diffusion into the HETCOR experiment. Comparison with data for a synthetic polymer, polycarbonate, indicates that the separation between O-alkyl and aromatic groups in the humic acid is less than 1.5 nm. However, transverse 13C relaxation filtering under 1H decoupling reveals heterogeneity on a nanometer scale, with the slow-relaxing component being rich in lignin-like aromatic-C-O-CH3 moieties and poor in COO groups.     

Microscale analysis of in vitro anaerobic degradation of lignocellulosic wastes by rumen microorganisms

Anaerobic degradation of lignin in waste straw by ruminal microbes was directly observed using atomic force microscope (AFM). A series of high-resolution AFM images of the straw surface in the biodegradation show that the wax flakelets and lignin granules covering the straw surface were removed by the rumen microorganisms. Such degradation resulted in an exposure of cellulose fibers located inside the straw. The appearance of holes and microfibers in fermentation reveals that tunneling might be one of the ways for rumen microorganisms to attack the straw. Increases in the atomic ratio of oxygen to carbon (O/C) and the ratio C2/C3 in C1s spectra of X-ray photoelectron spectroscopy confirm that more cellulose was exposed on the surface after the anaerobic fermentation of straw. Gas chromatography/mass spectrometry analytical results demonstrate the decomposition of lignin by rumen microorganisms. Fourier transform infrared spectroscopy spectra and the measurement of degradation efficiency of the main straw components further verify these microscaled observations.     

不同Cu离子浓度和pH值对云芝降解秸秆中木质素和纤维素的影响

云芝(Coriolus versicolor)对秸秆中木质素与纤维素的降解情况受不同Cu2+浓度和pH值条件的影响,设定不同条件可以提高木质素的降解率,降低纤维素的降解率,以更好地保存纤维素的完整性,实现秸秆中主要成分的分离。通过均匀设计法,设定不同的Cu2+浓度环境X1(0、1.0、2.0、4.0、6.0mmol/L)和不同pH值条件X2(2.2、2.8、3.4、4.0、4.6),测定相应条件下云芝的菌丝生长情况、木质素酶活力、纤维素酶活力以及秸秆中木质素和纤维素的降解率。结果表明：木质素的降解率与Cu2+浓度呈正相关,与pH值呈负相关;纤维素的降解率与pH值呈正相关,与Cu2+ 浓度呈负相关。通过试验所得方程预测得知,Cu2+浓度6mmol/L、pH2.2是优化的云芝降解、分离秸秆中主要成分的条件,在该条件下,由所得方程可知秸秆中木质素的降解率为16.1%,纤维素的降解率为27.7%。     

Importance of structural makeup of biopolymers for organic contaminant sorption

Sorption of pyrene, phenanthrene, naphthalene, and 1-naphthol by original (lignin, chitin, and cellulose) and coated biopolymers was examined. Organic carbon normalized distribution coefficients (Koc) of all compounds by the original biopolymers followed the order lignin > chitin > cellulose, in line with the order of their hydrophobicity. Hydrophobicity of structurally similar organic compounds is the main factor determining their ability to occupy sorption sites in biopolymers. Specific interactions (e.g., H-bonding) between 1-naphthol and chitin or cellulose increased its ability to occupy sorption sites. Lignin coating resulted in an increased Koc for phenanthrene (13.6 times for chitin and 6.9 times for cellulose) and 1-naphthol (6.0 times for chitin and 3.7 times for cellulose) relative to the acetone-treated chitin and cellulose. Also, these coated biopolymers had increased isotherm nonlinearity, due to the newly formed condensed domains. An increase in phenanthrene and 1-naphthol sorption by lignin-coated biopolymers as compared to chitin and cellulose was contributed by the newly created high-energy sites in condensed domains and coated lignin. Results of this study highlight the importance of the structural makeup of biopolymers in controlling the sorption of hydrophobic organic compounds.     

阳离子淀粉在磨木浆中的应用

从阳离子淀粉对化学浆、磨木浆的作用机理分析，阐明阳离子淀粉在磨木浆中应用成功的关键是要避开其木素等非纤维素物质的干扰。实践表明ＣＣＳ－０２型阳离子淀粉是提高新闻纸Ｚ向强度、印刷适性和助留、助滤的优良助剂。     

Multiple fluorescence labeling and two dimensional FTIR-13C NMR heterospectral correlation spectroscopy to characterize extracellular polymeric substances in biofilms produced during composting

Knowledge on the structure and function of extracellular polymeric substances (EPS) in biofilms is essential for understanding biodegradation processes. Herein, a novel method based on multiple fluorescence labeling and two-dimensional (2D) FTIR-(13)C NMR heterospectral correlation spectroscopy was developed to gain insight on the composition, architecture, and function of EPS in biofilms during composting. Compared to other environmental biofilms, biofilms in the thermophilic (>55 °C) and cooling (mature) stage of composting have distinct characteristics. The results of multiple fluorescence labeling demonstrated that biofilms were distributed in clusters during the thermophilic stage (day 14), and dead cells were detected. In the mature stage (day 26), the biofilm formed a continuous layer with a thickness of approximately 20-100 μm around the compost, and recolonization of cells at the surface of the compost was easily observed. Through 2D FTIR-(13)C NMR correlation heterospectral spectroscopy, the following trend in the ease of the degradation of organic compounds was observed: heteropolysaccharides > cellulose > amide I in proteins. And proteins and cellulose showed significantly more degradation than heteropolysaccharides. In summary, the combination of multiple fluorescence labeling and 2D correlation spectroscopy is a promising approach for the characterization of EPS in biofilms.     

Evaluation of pretreatment with Pleurotus ostreatus for enzymatic hydrolysis of rice straw

The effects of biological pretreatment of rice straw using four white-rot fungi (Phanerochaete chrysosporium, Trametes versicolor, Ceriporiopsis subvermispora, and Pleurotus ostreatus) were evaluated on the basis of quantitative and structural changes in the components of the pretreated rice straw as well as susceptibility to enzymatic hydrolysis. Of these white-rot fungi, P. ostreatus selectively degraded the lignin fraction of rice straw rather than the holocellulose component. When rice straw (water content of 60%) was pretreated with P. ostreatus for 60 d, the total weight loss and the degree of Klason lignin degraded were 25% and 41%, respectively. After the pretreatment, the residual amounts of cellulose and hemicellulose were 83% and 52% of those in untreated rice straw, respectively. By enzymatic hydrolysis with a commercial cellulase preparation for 48 h, 52% holocellulose and 44% cellulose in the pretreated rice straw were solubilized. The net sugar yields based on the amounts of holocellulose and cellulose of untreated rice straw were 33% for total soluble sugar from holocellulose and 32% for glucose from cellulose. The SEM observations showed that the increase in susceptibility of rice straw to enzymatic hydrolysis by pretreatment with P. ostreatus is caused by partial degradation of the lignin seal. When the content of Klason lignin was less than 15% of the total weight of the pretreated straw, enhanced degrees of enzymatic solubilization of holocellulose and cellulose fractions were observed as the content of Klason lignin decreased.     

Linking phosphorus sequestration to carbon humification in wetland soils by 31P and 13C NMR spectroscopy

Phosphorus sequestration in wetland soils is a prerequisite for long-term maintenance of water quality in downstream aquatic systems, but can be compromised if phosphorus is released following changes in nutrient status or hydrological regimen. The association of phosphorus with relatively refractory natural organic matter (e.g., humic substances) might protect soil phosphorus from such changes. Here we used hydrofluoric acid (HF) pretreatment to remove phosphorus associated with metals or anionic sorption sites, allowing us to isolate a pool of phosphorus associated with the soil organic fraction. Solution (31)P and solid state (13)C NMR spectra for wetland soils were acquired before and after hydrofluoric acid pretreatment to assess quantitatively and qualitatively the changes in phosphorus and carbon functional groups. Organic phosphorus was largely unaffected by HF treatment in soils dominated by refractory alkyl and aromatic carbon groups, indicating association of organic phosphorus with stable, humified soil organic matter. Conversely, a considerable decrease in organic phosphorus following HF pretreatment was detected in soils where O-alkyl groups represented the major fraction of the soil carbon. These correlations suggest that HF treatment can be used as a method to distinguish phosphorus fractions that are bound to the inorganic soil components from those fractions that are stabilized by incorporation into soil organic matter.     

Structural changes in wheat straw components during decay by lignin-degrading white-rot fungi in relation to improvement of digestibility for ruminants

Structural changes and resulting in-vitro dry matter digestibility (IVDMD) were determined during the course of solid-state fermentation of wheat straw using the lignin-degrading white-rot fungi Sporotrichum pulverulentum, Pycnoporus cinnabarinus and Cyathus stercoreus. The first fungus grew very rapidly on straw but degraded hemicelluloses and cellulose non-selectively resulting in very low IVDMD increases (50% after 35% weight loss). P. cinnabarinus and C. stercoreus preferentially degraded hemicelluloses achieving high improvements in IVDMD (maximum increase of 63 and 94%, respectively) with limited dry weight losses (12 and 18% after 7 and 13 days, respectively). The three fungi exhibited some selectivity among the individual hemicellulose components: O-acetyls were removed essentially at the same rate as xylan, while uronic acids accumulated as incubation proceeded. Conversely, the arabinose content decreased rapidly, especially with C. stercoreus and P. cinnabarinus, suggesting that removal of this pentose was partly responsible for digestibility improvement. Esterified phenolic acids were rapidly degraded during the first stages of decay by all three fungi although P. cinnabarinus and C. stercoreus degraded ferulic acid faster than p-coumaric acid. Lignin was preferentially degraded compared to polysaccharides by all three fungi. The amount of lignin removed, as determined by Klason, correlated well with IVDMD improvement (r=?0.97), while acid detergent lignin (ADL) showed a lower correlation (r=-0.86). Acidolysis yields of decayed lignin pointed to preferential degradation of β-O-4 ether linked units by the fungi. Syringyl units were removed faster than guaiacyl units only after 5 to 10% weight loss was obtained.     

Cross-polarization/magic-angle spinning 13C nuclear magnetic resonance study of cellulose I-ethylenediamine complex

Complete assignments of the cross-polarization/magic-angle spinning (CP/MAS) 13C nuclear magnetic resonance (NMR) spectrum of the cellulose I-ethylenediamine (EDA) complex, which is the intermediate of the reaction from cellulose I to cellulose III(I), were performed. In this paper, we used the 13C-enriched cellulose that was biosynthesized by Acetobacter xylinum ATCC10245 strain from culture medium containing D-(2-13C), D-(3-13C), or D-(5-13C)glucose as a carbon source. After conversion into cellulose I-EDA complex by sufficient EDA treatment, the CP/MAS 13C NMR spectra of the 13C-enriched cellulose I-EDA complexes were measured. As a result, 13C resonance lines of each carbon of the cellulose moiety in the complex appeared as a singlet, suggesting that all glucose residues of the complex are magnetically equivalent. The difference in chemical shifts for each carbon between cellulose I-EDA and cellulose I(alpha), I(beta), and III(I), respectively, suggests that the conformation of the cellulose chains for cellulose I-EDA differs from that for cellulose I(alpha), I(beta), and III(I). In addition, fitting analysis of the 13C spectrum of Valonia cellulose I-EDA complex revealed that the complex contains one EDA molecule per two glucose residues in the cellulose chain.