Water-soluble Cellulose Acetate Research Articles

Reverse Gradient Distributions of Drug and Polymer Molecules within Electrospun Core-Shell Nanofibers for Sustained Release.

Core-shell nanostructures are powerful platforms for the development of novel nanoscale drug delivery systems with sustained drug release profiles. Coaxial electrospinning is facile and convenient for creating medicated core-shell nanostructures with elaborate designs with which the sustained-release behaviors of drug molecules can be intentionally adjusted. With resveratrol (RES) as a model for a poorly water-soluble drug and cellulose acetate (CA) and PVP as polymeric carriers, a brand-new electrospun core-shell nanostructure was fabricated in this study. The guest RES and the host CA molecules were designed to have a reverse gradient distribution within the core-shell nanostructures. Scanning electron microscope and transmission electron microscope evaluations verified that these nanofibers had linear morphologies, without beads or spindles, and an obvious core-shell double-chamber structure. The X-ray diffraction patterns and Fourier transform infrared spectroscopic results indicated that the involved components were highly compatible and presented in an amorphous molecular distribution state. In vitro dissolution tests verified that the new core-shell structures were able to prevent the initial burst release, extend the continuous-release time period, and reduce the negative tailing-off release effect, thus ensuring a better sustained-release profile than the traditional blended drug-loaded nanofibers. The mechanism underlying the influence of the new core-shell structure with an RES/CA reverse gradient distribution on the behaviors of RES release is proposed. Based on this proof-of-concept demonstration, a series of advanced functional nanomaterials can be similarly developed based on the gradient distributions of functional molecules within electrospun multi-chamber nanostructures.

Multiscale Computational Study of Cellulose Acetate-Water Miscibility: Insights from Molecularly Informed Field-Theoretic Modeling.

Cellulose acetate (CA), a prominent water-soluble derivative of cellulose, is a promising biodegradable ingredient that has applications in films, membranes, fibers, drug delivery, and more. In this work, we present a molecularly informed field-theoretic model for CA to explore its phase behavior in aqueous solutions. By integrating atomistic details into large-scale field-theoretic simulations via the relative entropy coarse-graining framework, our approach enables efficient calculations of CA's miscibility window as a function of the degree of substitution (DS) of cellulose hydroxyl groups with acetate side chains. This allows us to capture the intricate phase behavior of CA, particularly its unique miscibility at intermediate substitution, without relying on experimental input. Additionally, the model directly probes CA solution behavior specific to the relative DS at C2, C3, and C6 alcohol sites, providing insights for the rational design of water-soluble CA for diverse applications. This work demonstrates a promising integration of molecularly informed field theories, complementing wet-lab experimentation, for engineering the next-generation polymeric materials with precisely tailored properties.

Design of Sustainable Aluminium-Based Feedstocks for Composite Extrusion Modelling (CEM).

Additive manufacturing (AM) has become one of the most promising manufacturing techniques in recent years due to the geometric design freedom that this technology offers. The main objective of this study is to explore Composite Extrusion Modelling (CEM) with aluminium as an alternative processing route for aluminium alloys. This process allows for working with pellets that are deposited directly, layer by layer. The aim of the technique is to obtain aluminium alloy samples for industrial applications with high precision, without defects, and which are processed in an environmentally friendly manner. For this purpose, an initial and preliminary study using powder injection moulding (PIM), necessary for the production of samples, has been carried out. The first challenge was the design of a sustainable aluminium-based feedstock. The powder injection moulding technique was used as a first approach to optimise the properties of the feedstock through a combination of water-soluble polymer, polyethyleneglycol (PEG), and cellulose acetate butyrate (CAB) wich produces low CO2 emissions. To do this, a microstructural characterisation was carried out and the critical solid loading and rheological properties of the feedstocks were studied. Furthermore, the debinding conditions and sintering parameters were adjusted in order to obtain samples with the required density for the following processes and with high geometrical accuracy. In the same way, the printing parameters were optimised for proper material deposition.

Cellulose eutecticgel electrolytes based on ethylene glycol/zinc chloride deep eutectic solvent for flexible solid-state capacitors

In the present work, we successfully prepared a conductive eutecticgel with good mechanical properties, adhesion properties and excellent antifreeze properties by mixing ethylene glycol (EG) as the hydrogen bonding donor and zinc chloride (ZnCl2) as the hydrogen bonding acceptor in different molar ratios to form an anhydrous deep eutectic solvent (DES). Water-soluble cellulose acetate (WSCA), monomer acrylamide (AM), initiator ammonium persulfate (APS) and cross-linking agent MBA (N, N′-methylene bis(acrylamide)) were added into the prepared DES to form a eutecticgel by a simple one-pot cooking strategy. The mechanical properties of the eutecticgel were strengthened by cross-linking of WSCA and PAM chains through hydrogen bonding, and the addition of the DES and ZnCl2 increased the freezing-resistant property of the eutecticgel. The stresses of DES-WSCA/PAM eutecticgel increased with the content of WSCA. The stress of DES-WSCA/PAM eutecticgel gradually becomes larger with the increase of WSCA content, and it can reach 184.07 kPa at 2 % WSCA content. In addition, the capacitor prepared by DES-WSCA/PAM eutecticgel shows excellent performance, with a voltage window of 2 V and a capacitance of 371.5 F/g, which can be operated stably for a long period, and the capacitance can still reach 145.4 F/g even under the condition of −40 °C. Moreover, DES-WSCA/PAM eutecticgel capacitor has a long service life, its capacitance retention rate and coulombic efficiency can reach more than 91 % and 95 %, respectively, after 10,000 times of charging and discharging tests. In summary, DES-WSCA/PAM eutecticgel electrolyte is an ideal material for capacitors, which opens up a new way of thinking for the development of multifunctional cellulose-based eutecticgels with low-temperature resistance, good mechanical properties, and non-irritating for wearable energy storage devices.

Tough conductive cellulose organohydrogels with antifreezing capability, strain-sensitivity and for multi-functional sensors

The mechanical strength of traditional hydrogels is weak, and it is easy to freeze and harden below zero to lose the characteristics and conductivity of hydrogels, which seriously limits the application of hydrogels in sensors. To solve this problem, an organic hydrogel was prepared using a simple method. After testing, this organic hydrogel not only has good frost resistance but also has excellent mechanical properties. Cellulose-based hydrogels have received increasing attention as good materials for preparing flexible, wearable sensors due to their good biocompatibility, flexibility, and excellent mechanical strength. We polymerized 4-carboxy benzaldehyde (FBA) and polyvinyl alcohol (PVA) into gels under acid-catalyzed conditions, imparted freeze resistance to the hydrogels by introducing dimethyl sulfoxide (DMSO) into the hydrogels, and added water-soluble cellulose acetate (WSCA) to the hydrogels to enhance the mechanical properties of the hydrogels. The organic hydrogel prepared by this method is more stable than conventional hydrogel in low-temperature environments, even below − 80゜C can still maintain the soft state and good mechanical flexibility of the hydrogel, of which WSCA can enhance the mechanical properties, and the stress of the hydrogel can reach 0.85 MPa when the WSCA content is 2%, which is 2.5 times that of the hydrogel without WSCA. In addition, the prepared hydrogel also has good electrochemical performance, under the condition of − 80゜C, connected to the circuit, the small bulb can still maintain a weak brightness. Moreover, the sensor composed of hydrogels can distinguish vigorous movement and weak movement of the human body, which provides a new idea for the application of hydrogels on the sensor.

Self-adhesive frost-resistant conductive hydrogel electrolytes based on TA@WSCA-Zn autocatalytic system for flexible and foldable solid-state capacitors

In this study, we have successfully prepared a freeze-resistant conductive hydrogel with a fast polymerization rate and excellent mechanical and adhesive properties. In this paper, hydrogels capable of rapid polymerization in a short time were prepared based on the TA@WSCA-Zn (WSCA, water-soluble cellulose acetate; TA@WSCA, tannic acid-coated WSCA suspension; TA@WSCA-Zn, a certain amount of zinc chloride (ZnCl2) was added to TA@WSCA suspension;) autocatalytic system at room temperature or low temperature. TA@WSCA-Zn forms a stable reversible quinone-catechol redox reaction to activate the decomposition of ammonium persulfate (APS, initiator) to produce SO4−· radicals, which triggers the ultra-fast polymerization of amphoteric ([2-(methacryloyloxy) ethyl] dimethyl-(3-sulfopropyl), SBMA) and acrylic monomers (AA). To improve the frost resistance and electrical conductivity of the hydrogels, we also introduced amphoteric ions, which can provide migration channels for Zn2+ and greatly increase the migration rate. The TA@WSCA-7.5/PSA hydrogel (PSA, hydrogel prepared by polymerization of acrylic acid and SBMA as monomers;) can achieve tensile strength and toughness of 1.33 MPa and 4.89 MJ m−3. It can provide excellent electrical conductivity and interfacial adhesion over a temperature range of 25 to −60 °C. In addition, the supercapacitor assembled from the TA@WSCA-7.5/PSA hydrogel electrolyte still has good capacitance characteristics after folding and 10,000 charge/discharge cycles, with a capacitance retention of 93.33 % and an excellent cycle life. This autocatalytic system provides a new strategy and approach for the preparation of freeze-resistant conductive hydrogel electrolytes.

Chemical and microbial characterization for fermentation of water-soluble cellulose acetate in human stool cultures.

Water-soluble cellulose acetate (WSCA), a synthetic fiber source, was applied to human stool cultures and to pure cultures of representative Bacteroides species to characterize the fermentation properties of WSCA in the human gut, and to assess the potential availability of WSCA as a food or additive candidate. All nine of the different types of WSCA tested here provided increased acetate levels in human stool cultures. Greater levels of deacetylation were observed as the degree of substitution of hydroxyl groups by acetyl groups decreased. Among the nine tested types of WSCA, CA-0.78-128 caused the largest shifts of the microbial community, including an increased abundance of members of the genus Bacteroides, especially Bacteroides uniformis. Of four representative human gut Bacteroides species, only B. uniformis grew in pure culture on WSCA to produce acetate actively. Water-soluble cellulose acetate has the potential for dietary application in human and other monogastric animals, based on the enhanced production of short-chain fatty acids (SCFAs), in particular acetate, in the hindgut. Short-chain fatty acid production is caused by selective proliferation of specific gut bacteria belonging to the genus Bacteroides. © 2020 Society of Chemical Industry.

Adsorption of cefotaxim sodium on activated carbons of various origin

The adsorption of the cephalosporin antibiotic sodium cefotaxime (CEFOT) on two activated carbons in the temperature range 288–313 K was under study. Experimental charcoal AC obtained from hydrolysis lignin was established to be mesoporous. It demonstrates greater CEFOT adsorption activity than the commercially available charcoal OU-A made from wood and characterized by both micro- and mesoporous structure. The surface modification of both activated carbons by a water-soluble polyelectrolyte cellulose acetate sulfate in the form of sodium salt (Na-CAS) leads to an increase in specific adsorption. The multilayer model with saturation (MMS) was used for data analysis. As a result, the number of molecules on the adsorption center, the density of adsorption centers, the total number of formed layers and the energy characteristics of adsorbate – adsorbent interactions at the interface and adsorbate – adsorbate ones within the adsorption layer were estimated.

Partially Acetylated Cellulose Dissolved in Aqueous Solution: Physical Properties and Enzymatic Hydrolysis.

Cellulose acetate is one of the most important cellulose derivatives. The use of ionic liquids in cellulose processing was recently found to act both as a solvent and also as a reagent. A recent study showed that cellulose dissolution in the ionic liquid 1-ethyl-3-methylimidazoliumacetate (EMIMAc) mixed with dichloromethane (DCM) resulted in controlled homogenous cellulose acetylation; yielding water-soluble cellulose acetate (WSCA). This research investigated the properties of cellulose acetate prepared in this manner, in an aqueous solution. The results revealed that WSCA fully dissolves in water, with no significant sign of molecular aggregation. Its conformation in aqueous solution exhibited a very large persistence length, estimated as over 10 nm. The WSCA exhibited surface activity, significantly reducing the surface tension of water. Because of the molecular dissolution of WSCA in water, augmented by its amphiphilicity, aqueous solutions of WSCA exhibited an overwhelmingly high rate of enzymatic hydrolysis.

Direct Electrospinning of Cellulose Acetate onto Polyurethane Sheet and Effect of Its Saponification on Mechanical Properties

This article reports on direct electrospinning of water-soluble cellulose acetate (WSCA) and cellulose diacetate (CDA) onto a non-conductive synthetic polymer sheet such as polyurethane (PU) sheet to modify its surface morphology. The direct electrospinning was successfully performed after spraying an anti-static agent on the PU sheet. Furthermore, tight fixation of the fibers was achieved by spraying a dimethylformamide/ethanol (1/1) solution during the spinning. Finally, the cellulose acetate fibers on the PU sheet were converted to regenerated cellulose fibers by saponification. Changes in surface morphology were evaluated via tribology tests. The friction coefficient of the PU sheet significantly decreased with increases in the amount of deposited fiber, thereby suggesting that the PU sheet surface became smoother. After saponification, the friction coefficients of the sheets slightly increased, with the exception of the PU sheet with the increased amount of saponified WSCA fibers, and this was potentially attributed to changes in the elasticity upon saponification.

Tough and conductive bio-based artificial nacre via synergistic effect between water-soluble cellulose acetate and graphene

Inspired by the hierarchical structure of natural nacre, a binary artificial nacre with excellent mechanical performance, tensile strength and toughness was successfully constructed based on water-soluble cellulose acetate (WSCA) and reduced graphene oxide (rGO). The tensile strength and toughness of the hybrid films were markedly improved by the introduction of rGO, and the tensile strength and toughness of the rGO/WSCA composite films reached 499.3 MPa and 9.6 MJ/m3 (rGO-WSCA-III), respectively, which are approximately 5.0 and 5.3 times that of natural nacre. In addition, the hybrid rGO/WSCA films showed better flame retardancy and stability in water, and high electrical conductivity. In this study, we provide a strategy to design and fabricate exceptionally strong and tough artificial nacre, which has potential application in tissue engineering and as a humidity sensor.

Bacterial Fermentation of Water-Soluble Cellulose Acetate Raises Large-Bowel Acetate and Propionate and Decreases Plasma Cholesterol Concentrations in Rats.

We hypothesized that water-soluble cellulose acetate (WSCA) could be useful tool for the delivery of short-chain fatty acids to the large intestine. Rats were fed a control diet or a diet containing graded levels of WSCA for up to 21 days. Consuming WSCA dose-dependently increased large-bowel acetate and propionate concentrations through the bacterial fermentation. When WSCA was used as substrate, acetyl esterase activity in the cecal bacteria was detected solely in rats fed WSCA, in which the activity increased over time accompanied by an increased number of Bacteroides xylanisolvens. Consuming WSCA at a 4% level increased the goblet cell numbers and mucin contents in the cecum and lowered plasma cholesterol concentrations, which tended to correlate with the portal plasma concentrations of propionate. The results suggest that bacterial fermentation of WSCA is characterized by the greater production of acetate and propionate, which may contribute to the physiologic alterations.

Cellulose acetate, a new candidate feed supplement for ruminant animals: In vitro evaluations

Cellulose acetate (CA), a derivative of cellulose in which some hydroxyl groups are substituted with acetyl groups, was evaluated as a new cellulosic feed source for ruminants. In the present work, a series of in vitro studies was carried out to determine how CA supplementation affects rumen fermentation and microbiota. Batch culture studies were conducted to select the type of CA suitable for feed use and to define the optimal supplementation level. Rumen fluid from 2 Holstein cows was mixed with McDougall's buffer in test tubes into which grass hay and concentrate containing a fiber source [cellulose (control), water-soluble CA (WSCA), or insoluble CA] had been placed. Each fiber source was supplemented at 10% of total substrate. Tubes were incubated for 24 h to determine fermentation and microbial parameters. Then, the dose response of these parameters to different supplementation levels of WSCA (0, 7.5, 15, 22.5, and 30%) was tested in the same manner. We also operated a continuous culture system with WSCA supplementation and evaluated the effects on digestibility, fermentation, and microbial parameters. The supplementation level of WSCA was set at 15% of total feed. In batch culture studies, WSCA, but not insoluble CA, yielded dose-dependent increases in ruminal acetate levels. In the continuous culture system study, WSCA yielded increases in ruminal acetate levels and in the abundance of bacteria of the genus Prevotella, including Prevotella ruminicola. Dry matter digestibility and total gas production were not affected. These results suggest that WSCA supplementation at 15% of total feed yielded increased acetate levels without negatively affecting feed digestion; these effects may reflect activation of Prevotella species. As ruminal acetate is involved in milk fat synthesis, WSCA can be considered as a candidate feed additive suitable for dairy cattle.

Three-dimensional Layered Water-Soluble Cellulose Acetate/Polyacrylamide Composites with Ultrahigh Ductility and Stretchability

Water-soluble cellulose acetate (WSCA), one of the most important cellulose derivatives, possesses biocompatibility, biodegradability and broad chemical modifying capacities. In this work, highly polymerized WSCA was firstly synthesized and used as cross-linker to fabricate highly ductile, tough and resilient WSCA/polyacrylamide (PAM) composite hydrogels. The results showed that the WSCA/PAM nanocomposite hydrogels exhibited extraordinary toughness and ductility with a tensile strength of 297 kPa and elongation at break of about 4020%. The enhancement of mechanical properties and stretchability were due to the synergistic effect from the hydrogen bonding and physical entanglement between the composite matrixes. Under stretching conditions, hydrogen bonds and the dense entanglement between WSCA chains and PAM could dynamically break and rearrange to dissipate energy. At the same time, the filaments of PAM embedded in layered WSCA matrix became unfolded or fractured to dissipate energy and maintained the conformation of hydrogels. It was envisioned that the introduction of WSCA into polymeric matrix would generate a facile method to fabricate multiple layered hybrid hydrogel network and significantly widen the WSCA applications in the preparation of high performance supramolecular systems.

Synthesis of Highly Polymerized Water-soluble Cellulose Acetate by the Side Reaction in Carboxylate Ionic Liquid 1-ethyl-3-methylimidazolium Acetate.

In the present study, we describe a novel one-step method to prepare water-soluble cellulose acetate (WSCA) with higher degree of polymerization values (DP = 650–680) by in situ activation of carboxyl group in ionic liquid. First of all, cellulose was dissolved in 1-ethyl-3-methylimidazolium acetate (EmimAc) and reacted with dichloroacetyl chloride (Cl2AcCl) in order to make cellulose dichloroacetate. Under various conditions, a series of water soluble products were produced. Elemental analysis and NMR results confirmed that they were cellulose acetate with DS (degree of substitution) values in the range from 0.30 to 0.63. NMR studies demonstrated that Cl2AcCl reacted with acetate anion of EmimAc producing a mixed anhydride that acetylated cellulose. Other acylating reagents such as benzoyl chloride, chloroacetyl chloride can also work similarly. 2D NMR characterization suggested that 6-mono-O-acetyl moiety, 3,6-di-O-acetylcellulose and 2,6-di-O-acetyl cellulose were all synthesized and the reactivity of hydroxyl groups in anhydro-glucose units was in the order C-6>C-3>C-2. This work provides an alternative way to make WSCA, meanwhile, also services as a reminder that the activity of EmimAc toward carbohydrate as acylating reagents could be a problem, because the expected acylated products may not be resulted and recycling of this ionic liquid could also be difficult.

Homogeneous synthesis of Ag nanoparticles-doped water-soluble cellulose acetate for versatile applications

We report a facile and efficient approach for synthesis of well-dispersed and stable silver nanoparticles (Ag NPs) using water-soluble cellulose acetate (CA) as both reductant and stabilizer. Partially substituted CA with highly active hydroxyl groups and excellent water-solubility is able to reduce silver ions in homogeneous aqueous medium effectively. The synthesized Ag NPs were characterized by UV⿿vis spectroscopy, X-ray diffraction, X-ray photoelectron spectroscopy, transmission electron microscopy and energy dispersive X-ray spectroscope analysis. The as-prepared Ag NPs were well-dispersed, showing a surface plasmon resonance peak at 426nm. The resulted Ag NPs@CA nanohybrids exhibit high catalytic activity for the reduction of 4-nitrophenol to 4-aminophenol in the presence of NaBH4. Meanwhile, the nanohybrids are also effective in inhibiting the growth of bacterial. This environmentally friendly method promotes the use of renewable natural resources to prepare a variety of inorganic-organic materials for catalysis, antibacterial, sensors and other applications.

Water-soluble cellulose acetate from waste cotton fabrics and the aqueous processing of all-cellulose composites

The objective of this study is to explore the possibility of using waste cotton fabrics (WCFs) as low cost feedstock for the production of value-added products. Our previous study (Tian et al., 2014) demonstrated that acidic ionic liquids (ILs) can be highly efficient catalysts for controllable synthesis of cellulose acetate (CA) due to their dual function of swelling and catalyzing. In this study, an optimized “quasi-homogeneous” process which required a small amount of acidic ILs as catalyst was developed to synthesize water-soluble CA from WCFs. The process was optimized by varying the amounts of ILs and the reaction time. The highest conversion of water-soluble CA from WCFs reached 90.8%. The structure of the obtained water-soluble CA was characterized and compared with the original WCFs. Moreover, we demonstrate for the first time that fully bio-based and transparent all-cellulose composites can be fabricated by simple aqueous blending of the obtained water-soluble CA and two kinds of nanocelluloses (cellulose nanocrystals and cellulose nanofibrils), which is attractive for the applications in disposable packaging materials, sheet coating and binders, etc.

Trans‐α‐xylosidase and trans‐β‐galactosidase activities, widespread in plants, modify and stabilize xyloglucan structures

Cell-wall components are hydrolysed by numerous plant glycosidase and glycanase activities. We investigated whether plant enzymes also modify xyloglucan structures by transglycosidase activities. Diverse angiosperm extracts exhibited transglycosidase activities that progressively transferred single sugar residues between xyloglucan heptasaccharide (XXXG or its reduced form, XXXGol) molecules, at 16 μM and above, creating octa- to decasaccharides plus smaller products. We measured remarkably high transglycosylation:hydrolysis ratios under optimized conditions. To identify the transferred monosaccharide(s), we devised a dual-labelling strategy in which a neutral radiolabelled oligosaccharide (donor substrate) reacted with an amino-labelled non-radioactive oligosaccharide (acceptor substrate), generating radioactive cationic products. For example, 37 μM [Xyl-³H]XXXG plus 1 mM XXLG-NH₂ generated ³H-labelled cations, demonstrating xylosyl transfer, which exceeded xylosyl hydrolysis 1.6- to 7.3-fold, implying the presence of enzymes that favour transglycosylation. The transferred xylose residues remained α-linked but were relatively resistant to hydrolysis by plant enzymes. Driselase digestion of the products released a trisaccharide (α-[³H]xylosyl-isoprimeverose), indicating that a new xyloglucan repeat unit had been formed. In similar assays, [Gal-³H]XXLG and [Gal-³H]XLLG (but not [Fuc-³H]XXFG) yielded radioactive cations. Thus plants exhibit trans-α-xylosidase and trans-β-galactosidase (but not trans-α-fucosidase) activities that graft sugar residues from one xyloglucan oligosaccharide to another. Reconstructing xyloglucan oligosaccharides in this way may alter oligosaccharin activities or increase their longevity in vivo. Trans-α-xylosidase activity also transferred xylose residues from xyloglucan oligosaccharides to long-chain hemicelluloses (xyloglucan, water-soluble cellulose acetate, mixed-linkage β-glucan, glucomannan and arabinoxylan). With xyloglucan as acceptor substrate, such an activity potentially affects the polysaccharide's suitability as a substrate for xyloglucan endotransglucosylase action and thereby modulates cell expansion. We conclude that certain proteins annotated as glycosidases can function as transglycosidases.

Differences in enzymic properties of five recombinant xyloglucan endotransglucosylase/hydrolase (XTH) proteins of Arabidopsis thaliana

Xyloglucan endotransglucosylase/hydrolases (XTHs) are cell wall enzymes that are able to graft xyloglucan chains to oligosaccharides or to other available xyloglucan chains and/or to hydrolyse xyloglucan chains. As they are involved in the modification of the load-bearing cell-wall components, they are believed to be very important in the regulation of growth and development. Given the large number (33) of XTH genes in Arabidopsis and the overlapping expression patterns, specific enzymic properties may be expected. Five predominantly root-expressed Arabidopsis thaliana XTHs belonging to subgroup I/II were analysed here. These represent two sets of closely related genes: AtXTH12 and 13 on the one hand (trichoblast-enriched) and AtXTH17, 18, and 19 on the other (expressed in nearly all cell types in the root). They were all recombinantly produced in the yeast Pichia pastoris and partially purified by ammonium sulphate precipitation before they were subsequently all subjected to a series of identical in vitro tests. The kinetic properties of purified AtXTH13 were investigated in greater detail to rule out interference with the assays by contaminating yeast proteins. All five proteins were found to exhibit only the endotransglucosylase (XET; EC 2.4.1.207) activity towards xyloglucan and non-detectable endohydrolytic (XEH; EC 3.2.1.151) activity. Their endotransglucosylase activity was preferentially directed towards xyloglucan and, in some cases, water-soluble cellulose acetate, rather than to mixed-linkage β-glucan. Isoforms differed in optimum pH (5.0-7.5), in temperature dependence and in acceptor substrate preferences.

Mixed‐linkage β‐glucan : xyloglucan endotransglucosylase, a novel wall‐remodelling enzyme from Equisetum (horsetails) and charophytic algae

Mixed-linkage (1-->3,1-->4)-beta-d-glucan (MLG), a hemicellulose long thought to be confined to certain Poales, was recently also found in Equisetum; xyloglucan occurs in all land plants. We now report that Equisetum possesses MLG:xyloglucan endotransglucosylase (MXE), which is a unique enzyme that grafts MLG to xyloglucan oligosaccharides (e.g. the heptasaccharide XXXGol). MXE occurs in all Equisetum species tested (Equisetum arvense, Equisetum fluviatile, Equisetum hyemale, Equisetum scirpoides, Equisetum telmateia and Equisetum variegatum), sometimes exceeding xyloglucan endotransglucosylase (XET) activity. Charophytic algae, especially Coleochaete, also possess MXE, which may therefore have been a primordial feature of plant cell walls. However, MXE was negligible in XET-rich extracts from grasses, dicotyledons, ferns, Selaginella and bryophytes. This and the following four additional observations indicate that MXE activity is not the result of a conventional xyloglucan endotransglucosylase/hydrolase (XTH): (i) XET, but not MXE, activity correlates with the reaction rate on water-soluble cellulose acetate, hydroxyethylcellulose and carboxymethylcellulose, (ii) MXE and XET activities peak in old and young Equisetum stems, respectively, (iii) MXE has a higher affinity for XXXGol (K(m) approximately 4 microM) than any known XTH, (iv) MXE and XET activities differ in their oligosaccharide acceptor-substrate preferences. High-molecular-weight (M(r)) xyloglucan strongly competes with [(3)H]XXXGol as the acceptor-substrate of MXE, whereas MLG oligosaccharides are poor acceptor-substrates. Thus, MLG-to-xyloglucan grafting appears to be the favoured activity of MXE. In conclusion, Equisetum has evolved MLG plus MXE, potentially a unique cell wall remodelling mechanism. The prominence of MXE in mature stems suggests a strengthening/repairing role. We propose that cereals, which possess MLG but lack MXE, might be engineered to express this Equisetum enzyme, thereby enhancing the crop mechanical properties.