Regio- and stereoselective anomeric esterification of glucopyranose 1,2-diols and a facile preparation of 2-O-acetylated glucopyranosyl trichloroacetimidates from the corresponding 1,2-diols

A highly regio- and stereoselective anomeric esterification of 3-O-allyl (or benzyl, or benzoyl)-4,6-O-isopropylidene-alpha,beta-d-glucopyranose with acetyl chloride, or allyl chloroformate, or ethyl chloroformate gave the corresponding 2-OH, 1-beta-acetates or -carbonates in excellent yields. The 2-OH, 1-beta-acetates were readily converted to the corresponding 2-O-acetylated glucopyranosyl trichloroacetimidates by reaction with trichloroacetonitrile via base promoted acetyl migration, while the 2-OH, 1-beta-carbonates were good glycosyl acceptors for the synthesis of (1-->2)-linked oligosaccharides.     

Rhizopus delemar Lipase in Microemulsion-based Organogels: Reactivity and Rate-Limiting Study

The reactivity of Rhizopus delemar lipase immobilized in sodium bis(2-ethylhexyl) sulfosuccinate microemulsion-based organogels (MBGs) was investigated as a function of the water content in the gel. Gelatin was used as the gelling component of the MBGs. The maximal reaction rate of 6.5 μM s^-1 was obtained at a W_G (molar ratio of water to AOT in the MBG phase) value of 100. In the experimental W_G conditions, the reaction proceeded under an effectiveness factor of 0.02 to 0.04 giving the reaction-limiting regime.     

Solvent Effects on Equilibrium Position and Initial Rate of Lipase-catalyzed Esterification Reactions in Organic Solvents: Experimental Results and Prediction Capabilities

The solvent effect on the equilibrium position and the initial rate of esterification of 1-hexanol with acetic acid catalyzed by a lipase has been experimentally investigated. A variety of non-polar and polar solvents have been considered and the results obtained indicate that the solvent effect on the equilibrium conversion is very important compared to that for transesterification reactions. A theoretically sound methodology using the group-contribution UNIFAC model for the prediction of solvent effects on the equilibrium position of enzymatic reactions is presented and it is applied to the reaction of 1-hexanol with acetic acid as well as to a similar reaction from the literature. The results obtained are better than those from empirical methods proposed in the literature such as correlations with the octanol-water partition coefficient of the solvent, as well as the solubility of water in the solvent. Moreover, the proposed methodology can be used for the determination of the equilibrium constant of the reaction. For the prediction of the solvent effect on the initial rate of enzymatic reactions it is found that it is more accurately determined using the product of the activities of the reactants, which can be predicted by the UNIFAC model, than the octanol-water partition coefficient of the solvent or the solubility of water in the solvent.     

Activity and Stability Studies Of Mucor miehei Lipase Immobilized in Novel Microemulsion-based Organogels

Lipase from Mucor miehei was immobilized in bis-(2-ethylhexyl)sulfosuccinate sodium salt (AOT) as well as lecithin water-in-oil (w/o) microemulsion-based organogels (MBGs) formulated with biopolymers such as agar and hydroxypropylmethyl cellulose (HPMC), respectively. These lipase-containing MBGs prove to be novel solid-phase catalysts for use in organic media. Using these organogels at 25°C, various esterification reactions in non-polar solvents as well as in solvent free systems were possible. Apparent lipase activity was influenced to some extent by the nature and the concentration of biopolymers used. Lipase stability in such MBGs is much higher than that observed in w/o microemulsions. MBGs containing lipase functioned effectively in repeated batch syntheses of fatty esters. Kinetic studies have shown that ester synthesis catalyzed by immobilized lipase occurs via the Ping-Pong bi-bi mechanism in which only inhibition by excess of alcohol has been identified. Values of all kinetic parameters were determined.     

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.     

Esterification of streptol — a cyclitol derivative — by Candida rugosa lipase: influence of the acyl donor on regioselectivity

The influence of the nature of acyl donors on the regioselectivity of Candida rugosa lipase for the esterification of streptol — a cyclitol derivative — was investigated. Excellent regioselectivity for the formation of 3,7-disubstituted derivatives was observed for vinyl butyrate (100% 3,7-derivative, 68% yield) and vinyl propionate (100% 3,7-derivative, 46% yield) as acyl donors. In contrast, for vinyl methacrylate as acyl donor, a mixture of 71% 3,7-derivative and 29% 1,7-derivative was obtained. Varying the chain length, a certain dependency of regioselectivity on the acyl donor was observed, however, no logical correlation satisfying all cases was found. Mono-substituted streptol derivatives were obtained by employing Novozym 243.     

Influence of the Drying Technique of Silica Gels on the Enzymatic Activity of Encapsulated Lipase

Recent studies by Reetz et al. (Reetz, M.T., Zonta, A. and Simpelkamp, J. (1996a) Biotechnology and Bioengineering, 49, 527-534) have shown that the catalytic activity of lipase encapsulated in sol-gel materials, in esterification reactions, depends on many parameters such as the presence of hydrophobic groups grafted on the gel network and of an organic component in the gel network. In the present study, we have examined the effect of the gel pore texture which can be modified by varying the drying technique, for a given silica precursor composition and hydrolysis-condensation procedure. For a given mixture of two silane precursors, propyltri-methoxysilane and tetramethoxysilane, we compared the effects of the presence or absence of an organic component such as polyvinyl alcohol, in combination with drying either by evaporation which leads to the formation of xerogels, or by supercritical drying in CO₂ which leads to the formation of aerogels. For this last technique, the exchange of liquid is also an important step and its effect on the enzyme activity has been examined. The gel pore texture was characterized by nitrogen absorption according to the Brunauer Emmett and Teller method. The catalytic activities of the materials were compared in the esterification of lauric acid by 1-octanol.     

Selective oxidation-reduction and esterification of asiatic acid by Pestalotiopsis microspora and anti-HCV activity

Microbial transformation of asiatic acid (AA) by an endophytic fungus, Pestalotiopsis microspora, yielded six metabolites: 2-oxo-3β,15α,23-trihydroxyurs-12-ene-28-oic acid (1); 2-oxo-3β,15α,22α,23-tetrahydroxyurs-12-ene-28-oic acid (2); 2-oxo-3β,15α,23,30-tetrahydroxyurs-12-ene-28-oic acid (3); 2α,3β,15α,23,30-pentahydroxyurs-12-ene-28-oic acid methyl ester (4); 2α,3α,15α,23-tetrahydroxyurs-12-ene-28-oic acid (5); 2α,3α,15α,23,30-pentahydroxyurs-12-ene-28-oic acid (6). The structure elucidation of these products was confirmed based on the spectroscopic data. Compounds 2–6 were new. A possible biotransformation pathway is proposed. The anti-HCV activity of compounds 1–6 was also evaluated.     

Ultrasonic pretreatment for lipase-catalyzed synthesis of 4-methoxy cinnamoyl glycerol

4-Methoxy cinnamoyl glycerol (4MCG) is a very promising UV filters material in personal care products. In order to effectively improve the yield of 4MCG, a systematic study on ultrasonic pretreatment enzymatic esterification for 4MCG products was carried out. An ultrasonic frequency of 35 kHz, ultrasonic power of 150 W and ultrasound irradiation time of 1.5 h was determined to guarantee satisfactory degree of esterification and lipase activity. The optimum production was achieved in organic solvent system at 65 °C with 4MCA to glycerol molar ratio of 1:5, enzyme amount of 15 mg/mL, resulting in a monoester yield of above 66% and 55% after 48 h and 24 h of reaction under ultrasonic pretreatment, respectively. The experimental kinetic data were studied. The reactions were modeled by a system of sequential first-order rate expressions, kinetic parameters were estimated from experimental data fit to the model equations. Results show that the monoester yield in the ultrasonic pretreatment process (24 h) were above 1.5-fold as that in mechanical stirring process without essential damaging to lipase activity. The enzymatic method using ultrasonic pretreatment was obviously superior to the mechanical stirring for enzymatic method and chemical method in terms of conversion rate and the monoester yield. These results are of great significance for applying ultrasonic pretreatment method to prepare 4MCG.