Takao Raku

Selective enzymatic preparation of vinyl sugar esters using DMSO as a denaturing co-solvent

The protease-catalyzed transesterifications between hexoses and divinyladipate were examined. In dimethylformamide hexoses such as d-glucose, d-mannose, d-galactose and α-methyl d-galactoside were esterified with divinyladipate by alkaline protease from Streptomyces sp. to give corresponding 6-O-vinyl adipoyl sugars. When the denaturing cosolvent, DMSO, was added to the solvent, galactose was selectively esterified at only the C-2 position.

Enzymatic synthesis of trehalose esters having lipophilicity.

To improve trehalose lipophilicity, trehalose was regioselectively esterified with vinyl fatty acid esters in dimethyl formamide by protease from Bacillus subtilis to give 6-O-lauroyltrehalose, 6-O-myristoyltrehalose, 6-O-palmitoyltrehalose, 6-O-stearoyltrehalose, 6-O-oleoyltrehalose and 6-O-linoleoyltrehalose. The influence of structural variation by changing fatty acid substitute was examined by measurement of the surface tension and biodegradability.

Enzymatic synthesis of hydrophilic undecylenic acid sugar esters and their biodegradability.

To enhance water solubility of 10-undecylenic acid, which has anti-fungus, anti-bacterial and anti-virus activity, d-glucose, trehalose and sucrose were regioselectively esterified with vinyl 10-undecylenic acid ester in dimethyl formamide by a commercial protease, Bioprase conc., from Bacillus subtilis. 6-O-(10-Undecylenoyl) d-glucose, 6-O-(10-undecylenoyl) trehalose and 1′-O-(10-undecylenoyl) sucrose were obtained. The influence of structural variation by changing the sugar moiety was analyzed the surface tension and biodegradability.

Preparation of vinyl thymidine ester catalyzed by protease and its chemical polymerization

Transesterification reaction of 0.25 M thymidine with 1 M divinyladipate in dimethylformamide (DMF) was catalyzed by an alkaline protease (5 mg ml−1) from Streptomyces sp. (20 units mg−1 min) at 30 °C for 7 days to give 5′-O-vinyladipoyl thymidine (yield 77%) without formation of any by-products. Poly(vinyl alcohol) containing thymidine branches could be obtained by its free-radical polymerization.

Synthesis of vinyl arabinose ester catalyzed by protease from Streptomyces sp.

The transesterification of 0.5 M divinyladipate with 0.25 M arabinose in dimethylformamide for 7 days was catalyzed by Streptomyces sp. alkaline protease to give 5-O-vinyladipoyl-d-arabinofuranose at ca. 50% yield. Only enzymatic transesterification of primary hydroxyl group of arabinofuranose proceeded without esterification of arabinopyranose.

Synthesis of thermosensitive polymers containing sugar branches

Copolymers of 6-O-vinyladipoyl-D-glucose (VAG) and N-isopropyl acrylamide (NIPAm) were synthesized by radical polymerization. The number-average molecular weights of the copolymers were 3 × 104 ≈ 6 × 104. The observed segment composition of copolymers at the feed molar ratio (VAG 25/NIPAm 75) was VAG 10/NIPAm 90. The polymerization rate of the VAG monomer was slower than that of the NIPAm monomer. The lower critical-solution temperature of copolymers measured with a light-scattering photometer and a differential scanning calorimeter increased with increasing VAG segment composition. The increase in transition temperature was accompanied by a decrease in transition heat.

Transesterification of divinyladipate with glucose at various temperatures by an alkaline protease of Streptomyces sp.

The transesterification of 1 M divinyladipate with 0.25 M glucose in dimethylformamide (DMF) catalyzed by 5 mg ml−1 alkaline protease (24 units mg−1 min−1) from Streptomyces sp. gave 6-O-vinyladipoyl d-glucose as the main product with yields are between 60 and 90%. The optimum temperature for the reaction was about 50 °C.

Regio-selective preparation of vinyl adenosine ester catalyzed by alkaline protease

The transesterification of divinyladipate with adenosine in DMF containing 20% (v/v) DMSO was catalyzed by Streptomyces sp. alkaline protease and esterification occurred exclusively at the 3′-position of hydroxyl group of ribofuranose in adenosine to give 3′-O-vinyladipoyl adenosine without other products.

Lipase catalyzed reaction of L-ascorbic acid with cinnamic acid esters and substituted cinnamic acids

To perform the lipase-catalyzed synthesis of L-ascorbic acid derivatives from plant-based compounds such as cinnamic and ferulic acid under mild reaction conditions, the activities of immobilized Candida ntarctica lipase with different cinnamic acid esters and substituted cinnamic acids were compared. As a result, immobilized C. ntarctica lipase was found to prefer vinyl cinnamic acid to other esters such as allyl-, ethyl-, and isobutyl cinnamic acids as well as substituted cinnamic acids such as p-coumaric acid, caffeic acid, ferulic acid, and sinapic acid. Based on these results, large-scale synthesis of 6-O-cinnamyl-L-ascorbic acid ester was performed using immobilized C. ntarctica lipase in dry organic solvent, resulting in 68% yield (493 mg) as confirmed by 13C-NMR.

Synthesis, characterization and evaluation of new polymers for thermo- precipitation of adenosine

N-Isopropylacrylamide (NIPAM) and O-vinyladipoyl uridine were copolymerized in different feed molar ratios ca. 95:5 to 75:25, respectively. The resulting polymers were characterized by gel-permeation chromactography, differential scanning calorimeter and cloud point measurements. The number-average molecular weights of the copolymers were 1.6×104≈3.3×104. The lower critical solution temperature of the copolymer of NIPAM and uridine (95:5) decreases up to 29 °C, compared to that of poly(NIPAM) which remains at 33 °C. These polymers exhibited greater than 80% precipitation of adenosine with 60% efficiency of recovery.