The oxidation and (methylthio)methylation of cotton by methyl sulfoxide-acetic anhydride

Methyl sulfoxide-acetic anhydride oxidizes some of the cellulosic hydroxyl groups of cotton yarn to aldehyde and ketone groups, while etherifying other hydroxyl groups with (methylthio)methyl groups. The ratio of oxidation to (methylthio)methylation of the cellulose is influenced by the reaction temperature, the reaction time, and the ratio of methyl sulfoxide to acetic anhydride employed. The (methylthio)methyl groups may be cleaved by boiling the cotton in aqueous solutions of acetic acid.The total carbonyl content of the oxidized cellulose was determined by conversion into the polyoxime. The ratio of aldehyde to ketone groups in the oxidized cellulose was determined by further oxidation with chlorous acid; the car?yl groups produced were measured spectrophotometrically by a method involving adsorption of Methylene Blue.     

New cellulose sulfate derivatives and applications

The preparation of water-soluble cellulose sulfates having a degree of substitution (d.s.) of 0.3–2.0 by a homogeneous method is reported. Instead of sulfating cellulose directly, a reactive intermediate, namely, cellulose nitrite, was used, thereby permitting sulfation to proceed under homogeneous conditions. Products obtainable via the homogeneous method differ substantially from those prepared via the heterogeneous method used at present for the production of water-soluble cellulose derivatives. Principal chemical differences are the uniform distribution of the substituents, the exceptionally high molecular weight of the sulfate esters, and in the location of the substituents on the glucosyl residues. The differences result in a number of chemical and physical properties that are unique and not exhibited by other cellulose derivatives. Performance in application areas, such as oil recovery, paints, paper, textiles, cosmetics, and the like, is indicated by these properties.     

N-乙酰化反应制备水溶性壳聚糖研究

将高度脱乙酰化的壳聚糖在均相介质中进行N-乙酰化反应,制备水溶性壳聚糖。研究了制备工艺条件对脱乙酰度及水溶性的影响。结果表明,在乙酸—乙醇均相体系中进行乙酰化反应时,壳聚糖与乙酸酐的质量比为1∶0.6,反应温度控制在20℃,反应时间为8 h时,产品的脱乙酰度在50%左右,获得了水溶性良好的N-乙酰化壳聚糖。     

氨基酸的消旋研究

在纯乙酸溶剂中,以乙酸酐酰化消旋酪氨酸,消旋温度为90℃,L-酪氨酸和乙酸酐的摩尔比为12,每消旋1gL-酪氨酸时,溶剂乙酸的用量为5mL,效果较好。同样的方法应用于L-脯氨酸,L-苏氨酸,L-缬氨酸,L-赖氨酸,L-半胱氨酸等氨基酸,消旋率均可达到100%。     

Location of sulfate groups on sulfoacetate derivatives of cellulose

A water-soluble cellulose acetate sulfate (CAS) with a degree of acetylation (DS(Ac)) 2.4 and a degree of sulfation (DS(Sulf)) of 0.3 was obtained by direct acetylation of cellulose using sulfuric acid as catalyst. Using methylation analysis, IR and NMR spectroscopy, sulfate groups have been located on primary alcohol function of glucose residues. The distribution of the sulfate groups along the cellulose chain has been investigated using enzymatic hydrolysis. CAS was first de-acetylated under mild hydrolysis conditions (NaOH 0.25 mol/L at room temperature), and then cellulose sulfate was hydrolyzed by a cellulolytic complex (Celluclast 1.5L). Reaction products were separated by ion exchange chromatography on a DEAE Sepharose CL6B column into five fractions F(1), F(2), F(3), F(4) and F(5), which were analyzed for their chemical composition. F(1) was glucose and represented the main product of reaction (approximately 50% of the initial glucose), F(2) was a dimer (approximately 30%) with a ratio Sulfates-Glucose of 0.41 (about one sulfate group for two glucose units), F(3) a trimer (approximately 10%) with a ratio Sulfates-Glucose of 0.62 (about two sulfate groups for three glucose units), and F(4) a tetramer (approximately 5%) with a ratio Sulfates-Glucose of 0.69. The structure of the oligomers was established using 1H and 13C NMR. The observed proportion of the different blocks of sulfate groups was in good agreement with computed random distribution.     

Esterification of cellulose-enriched agricultural by-products and characterization of mechanical properties of cellulosic films

Two agricultural by-products, wheat bran and maize bran have been examined for their suitability to be transformed into biomaterials by esterification by lauroyl chloride. Influence of biochemical characteristic of cellulose (cellulose content, viscosity-average degree of polymerization, crystallinity) was studied on eight samples enriched in cellulose after chemical removal of heteroxylans and lignin. After an acidic pre-treatment, esterification was carried out with lauroyl chloride and an optimized reaction time of 8 h was used.

Chemical compositions were similar for all cellulose esters obtained, but cellulose content of initial material had a marked influence on the amount of esterified product. A film was easily obtained by casting and the mechanical (tensile strength and elongation), thermomechanical and calorimetric properties were determined. The possible role of grafted fatty acid as internal plasticizer was finally discussed.     

The role of water-soluble components in phytotoxicity from decomposing straw

Summary When seedling development is slowed by the presence of straw in wet seed-beds both microbial products and compounds of plant origin contribute to phytotoxicity. Hot (100°C) water-soluble extracts from fresh straw contained phytotoxic substances but these accounted for less of the phytotoxicity than the microbial products, primarily acetic acid, from anaerobic fermentation of the insoluble straw polysaccharides (cellulose and hemicelluloses). The water-soluble components however also included mineral salts required in the decomposition of these polysaccharides.     

Immobilization of BSA, enzymes and cells of Bacillus stearothermophilus onto cellulose, polygalacturonic acid and starch based graft copolymers containing maleic anhydride

Poly(maleic anhydride styrene) graft copolymers of cellulose, pectin polygalacturonic acid salt, calcium polygalacturonate, and starch were prepared and used to immobilize proteins. The cellulose grafts coupled quite appreciable quantities of acid phosphatase, glucose oxidase, and trypsin. However, the general retention of activity was somewhat disappointing. Further investigation with acid phosphatase showed that the amount of enzyme immobilized increased as the amount of anhydride in the graft copolymer increased but no such relationship existed for the enzymic activity. The cellulose graft copolymers were hydrolyzed and it appeared that the carboxyl group aided adsorption of the enzyme. Attempts to couple acid phosphatase using CMC through the free carboxyl groups, created by hydrolysis, gave only a small increase in the extent of protein coupling. However, the unhydrolyzed system gave a useful degree of immobilization of cells of Bacillus stearothermophilus, as did a poly(maleic anhydride/styrene)-cocellulose system. Attempts to improve the activity by using grafts based on other polysaccharide supports met with mixed success. Pectin products were soluble. Polygalacturonic acid products were partially soluble and extremely high levels of enzymic activity were obtained. This was probably due in part to the hydrophilic nature of the system, which also encouraged absorption of the enzyme. Attempts were made to reduce the solubility by using the calcium pectinate salt. Immobilization of acid phosphatase and trypsin resulted in inceased protein coupling but relatively poor activities were attained. A starch based system gave similar results. Calcium polygalacturonate was used to prepare an insoluble graft copolymeric system containing acrylonitrile-comaleic anhydride. The resulting gels gave excellent coupling with acid phosphatase which had a very good retention of activity.     

A kinetic study of isoamyl acetate synthesis by immobilized lipase-catalyzed acetylation in n-hexane

The objective of this work was to propose a reaction mechanism and to develop a rate equation for the synthesis of isoamyl acetate by acylation of the corresponding alcohol with acetic anhydride using the lipase Novozym 435 in n-hexane. The reaction between isoamyl alcohol and acetic anhydride occurred at high rate in first place. Then, if excess alcohol was used, produced acetic acid further reacted with remaining alcohol, leading to yields higher than 100% (based on initial acetic anhydride content). This reaction was much slower and took place only when acetic anhydride had been totally consumed. Optimal pH for Novozym 435 was 7.7. Acetic acid strongly inactivated the enzyme but it was partially caused by the pH drop in the biocatalyst aqueous microenvironment. Acetic anhydride also showed an important inhibition effect. On the contrary, isoamyl alcohol and isoamyl acetate had no negative effect on the lipase. The analysis of the initial rate data showed that reaction followed a Ping-Pong Bi-Bi mechanism with inhibition by acetic anhydride. The kinetic constants were obtained by multiple regression analysis of experimental findings. Equation predictions and experimental reaction rate values matched very well at conditions where acetic acid concentration in the medium was low.     

Qualitative analysis of products formed during the acid catalyzed liquefaction of bagasse in ethylene glycol

Bagasse was liquefied in ethylene glycol (EG) catalyzed by sulfuric acid at 190 degrees C under atmospheric pressure. The compositions of the crude products obtained were analyzed after separating them into three fractions: a water-soluble fraction, an acetone-soluble fraction and a residue. With infrared, gel permeation chromatography and elemental analyses, the residue mainly included undissolved cellulose and lignin derivatives and the acetone-soluble fraction mainly contained lignin degradation products with high molecular weights. The water-soluble fraction, after further analyzed by GC-MS and HPLC, showed EG, diethylene glycol, EG derivatives, saccharides, alcohols, aldehydes, ketones, phenols, especially some acids such as formic acid, levulinic acid, acetic acid, oxalic acid and 2-hydroxy-butyric acid and their esters. The Higher Heating Value (HHV) of the residue and the acetone-soluble fractions were higher than that of bagasse. The results showed that some useful chemicals and biofuels could be obtained by this process.     

The determination of purine levels in human and mouse plasma

Variable levels of acetic anhydride have been recommended for addition to one of two reagents used in the glyoxylic acid method for the determination of tryptophan. For use of this reagent immediately after preparation it was shown that a minimum of 16% (v/v) of acetic anhydride should be included in the formulation to obtain near-maximum sensitivity. It was further demonstrated that reagent formulations with and without acetic anhydride changed with exposure to light. The observed changes are manifest as changes in the relative sensitivities of the assay. Several modifications are recommended to improve the sensitivity and stability of the acetic anhydride-containing reagent in this assay.     

Clostridium lentocellum SG6--a potential organism for fermentation of cellulose to acetic acid

A cellulolytic, acetic acid producing anaerobic bacterial isolate, Gram negative, rod-shaped, motile, terminal oval shaped endospore forming bacterium identified as Clostridium lentocellum SG6 based on physiological and biochemical characteristics. It produced acetic acid as a major end product from cellulose fermentation at 37°C and pH 7.2. Acetic acid production was 0.67 g/g cellulose substrate utilized in cellulose mineral salt (CMS) medium. Yeast extract (0.4%) was the best nitrogen source among the various nitrogenous nutrients tested in production medium containing 0.8% cellulose as substrate. No additional vitamins or trace elemental solution were required for acetic acid fermentation. This is the highest acetic acid fermentation yield in monoculture fermentation for direct conversion of cellulose to acetic acid.     

Microfingerprints of proteins through labeling acetylation of their proteolytic peptides

Microgram amounts of proteins applied to polyacrylamide gel electrophoresis were subjected to a fingerprinting procedure using a combined proteolysis-acetylation method with the aid of ¹⁴C-labeled acetic anhydride of high specific activity. After staining, gel slices were partially dried and were resoaked in a solution of a protease. After elution and acetylation, the resulting peptides were resolved in fingerprints on cellulose thin-layer chromatography plates and subjected to autoradiography with or without sensitization. Yields, completeness of fingerprinting, and possible artefacts were investigated.     

Solvent-free process to esterify polysaccharides

A novel process for the preparation of acetates of polysaccharides is described herein. The process involves the acetylation of polysaccharides with acetic anhydride in the presence of iodine as a catalyst. No solvent is required to bring about the acetylation. The method is simple, rapid, and characterized by a high conversion ratio. Conversion of cellulose and starch into their corresponding acetate derivatives has been demonstrated.     

Fermentation of cellulose to acetic acid by Clostridium lentocellum SG6: induction of sporulation and effect of buffering agent on acetic acid production

AIMS: To determine the growth, correlation between sporulation and acetic acid production and effect of buffering agent at high substrate cellulose concentrations of the strain Clostiridium lentocellum SG6. METHODS AND RESULTS: The strain SG6 was grown in cellulose mineral salt medium containing cellulose (Whatman No. 1 filter paper, Whatmore International Ltd., Maidstone, UK) or cellobiose. The strain fermented cellulose even after several transfers on cellobiose medium. The formation of endospores on third day onwards indicated the lowering of pH in the medium because of the formation of acetic acid. Maintaining the pH 7.2 at higher substrate concentrations resulted in increase of biomass, cellulose fermentation, acetic acid production, etc. CONCLUSIONS: The strain SG6, with its high fermentation yields and sporulating character can become a potential strain for acetic acid production and also as a probiotic strain in animal nutrition. SIGNIFICANCE AND IMPACT OF THE STUDY: The direct conversion of cellulosic biomass to acetic acid can eliminate expensive three-step saccharification, fermentation processes. The strain SG6 can ferment cellulose at high substrate concentrations.     

Homogeneous modification of sugarcane bagasse cellulose with succinic anhydride using a ionic liquid as reaction medium

The homogeneous chemical modification of sugarcane bagasse cellulose with succinic anhydride using 1-allyl-3-methylimidazolium chloride (AmimCl) ionic liquid as a reaction medium was studied. Parameters investigated included the molar ratio of succinic anhydride/anhydroglucose units in cellulose in a range from 2:1 to 14:1, reaction time (from 30 to 160min), and reaction temperature (between 60 and 110 degrees C). The succinylated cellulosic derivatives were prepared with a low degree of substitution (DS) ranging from 0.071 to 0.22. The results showed that the increase of reaction temperature, molar ratio of SA/AGU in cellulose, and reaction time led to an increase in DS of cellulose samples. The products were characterized by FT-IR and solid-state CP/MAS (13)C NMR spectroscopy, and thermal analysis. It was found that the crystallinity of the cellulose was completely disrupted in the ionic liquid system under the conditions given. The data also demonstrated that homogeneous modification of cellulose with succinic anhydride in AmimCl resulted in the production of cellulosic monoester. The thermal stability of the succinylated cellulose decreased upon chemical modification.     

Formation of brown lines in paper: characterization of cellulose degradation at the wet-dry interface

Brown lines were generated at the wet-dry interface on Whatman paper No. 1 by suspending the sheet vertically in deionized water. Formic acid and acetic acid were quantified in three areas of the paper defined by the wet-dry boundary (above, below, and at the tideline) using capillary zone electrophoresis with indirect UV detection. Their concentration increased upon accelerated aging of the paper and was highest in the tideline. The hydroperoxides have been quantified using reverse phase high performance liquid chromatography with UV detection based on the determination of triphenylphosphine oxide produced from the reaction with triphenylphosphine, and their highest concentration was found in the tideline as well. For the first time, it was shown that various types of hydroperoxides were present, water-soluble and non-water-soluble, most probably in part hydroperoxide functionalized cellulose. After accelerated aging, a significant increase in hydroperoxide concentration was found in all the paper areas. The molar masses of cellulose determined using size-exclusion chromatography with multiangle light scattering detection showed that, upon aging, cellulose degraded significantly more in the tideline area than in the other areas of the paper. The area below the tideline was more degraded than the area above. A kinetic study of the degradation of cellulose allowed determining the constants for glycosidic bond breaking in each of the areas of the paper.     

Fermentative Conversion of Cellulose to Acetic Acid and Cellulolytic Enzyme Production by a Bacterial Mixed Culture Obtained from Sewage Sludge

A simple procedure that uses a cellulose-enriched culture started from sewage sludge was developed for producing cellulolytic enzymes and converting cellulose to acetic acid rather than CH₄ and CO₂. In this procedure, the culture which converts cellulose to CH₄ and CO₂ was mixed with a synthetic medium and cellulose and heated to 80°C for 15 min before incubation. The end products formed were acetic acid, propionic acid, CO₂, and traces of ethanol and H₂. Supernatants from 6- to 10-day-old cultures contained 16 to 36 mM acetic acid. Cellulolytic enzymes in the supernatant were stable at 2°C under aerobic conditions for up to 4 weeks and had the ability to hydrolyze carboxymethyl cellulose, a microcystalline cellulose, cellobiose, xylan, and filter paper to reducing sugars.     

A spectrophotometric method of analysis of fusidic acid

A spectrophotometric method for assay of fusidic acid is described. The method is based on reaction with a reagent consisting of acetic anhydride and concentrated sulfuric acid. Mathematical processing of the results of the main substance determination in fusidic acid preparations showed that the error did not exceed 2 per cent. Procedures for spectrophotometric assay of fusidic acid in control of the processes of its biosynthesis, isolation and purification were developed. The procedures provided control of the technological process of fusidic acid production.     

Acetylation of pea isolate in a torus microreactor

Acetylation, which acts on the amino groups of proteins, allows to increase the solubility and the emulsifying properties of pea isolate. Acetylation by acetic anhydride was carried out in a torus microreactor in semibatch and continuous conditions. The mixing characteristics, obtained by a residence time distribution (RTD) method, are the same in batch and continuous processes. The maximum acetylation degree reached by the torus reactor is higher than with the stirred reactor. Torus reactors are more efficient than stirred ones as shown by a conversion efficiency, defined by the quantity of modified lysine groups by consumed acetic anhydride. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 53: 409-414, 1997.