Production of optically active esters and alcohols from racemic alcohols by lipase-catalyzed stereoselective transesterification in non-aqueous reaction system.

Microbial lipase-catalyzed transesterification between vinyl acetate and (RS)-2-octanol or (RS)-1-phenylethanol was investigated in a reaction system without addition of aqueous or organic solvents. From a screening test with various lipases, it was found that the enzymes from Pseudomonas species could efficiently catalyze the reaction, and R-enantiomers of the racemic alcohols were preferentially esterified by them. Enantiomeric purities of the optically active alcohols (S) and esters (R) obtained from (RS)-1-phenylethanol by the stereoselective transesterification of these lipases were all more than 95%.

Studies of the site and mode of biosynthesis of tobacco trichome exudate components.

Detached glandular trichome head preparations and epidermal strips with and without trichome heads were used to identify glandular trichome heads as the site of sucrose ester biosynthesis in tobacco. Carbon dioxide in solution as well as sucrose, glucose, and acetate were shown to serve as precursors to both sucrose esters and duvatrienediol diterpenes in detached trichome heads or epidermal strips, and gaseous CO2 was also efficiently utilized by epidermal strips. Thus, glandular heads can biosynthesize these principal exudate components from a molecule as simple as CO2. While formation of duvatriendiols from all precursors tested and conversion of sucrose and glucose to sucrose esters was light dependent, utilization of acetate to label the 6-O-acetyl group of the glucose moiety of sucrose esters occurred equally well in light and dark. The data suggest that CO2 and/or monosaccharides produced in trichome head cells and perhaps that supplied by other epidermal cells can act as carbon sources for sucrose ester and duvatrienediol biosynthesis which occurs in the glandular trichome head.

Formation of fatty acid ethyl esters during chronic ethanol treatment in mice.

Ethyl esters of long-chain fatty acids are formed in the liver and brain of mice after 1-6 days of ethanol intoxication. This observation extends the reports of Lange and co-workers who detected these compounds as unusual metabolites of ethanol in human tissues [E. A. Laposata and L. G. Lange, Science 231, 497 (1986)]. Ethyl esters of oleic and linoleic acids, and, in smaller amounts, ethyl esters of palmitic and stearic acids were found in the livers of mice that had been treated with ethanol by inhalation. In the brain, only the esters of unsaturated fatty acids were found, in lower amounts than in liver. All the fatty acid ethyl esters seemed to have reached steady-state levels in the tissues after 3 or 4 days of alcohol treatment. When incorporated into synaptosomal plasma membranes in vitro, in intramembrane concentrations estimated to resemble those observed in the mice, these esters reduced the fluorescence anisotropy, i.e. they disordered the membranes.

Ester synthesis at extraordinarily low temperature of -3 degrees C by modified lipase in benzene.

The lipoprotein lipase from Pseudomonas fluorescens was modified with 2,4-bis(O-methoxypolyethylene glycol)-6-chloro-s-triazine. The modified lipase in which 55% of the amino groups in the enzyme molecule were coupled with polyethylene glycol was found to be soluble in benzene and catalyzed the reactions of ester synthesis, ester exchange, aminolysis and ester hydrolysis in benzene. The modified lipase had an extraordinary temperature-dependency: enzymic activity for methyl laurate synthesis from methyl alcohol and lauric acid increased with decreasing temperature and attained the maximum at the extremely low temperature of -3 degrees C. The optimum temperature for hydrolysis of methyl laurate was as low as -4 degrees C.

Identification of fatty acid ethyl esters as products of rabbit myocardial ethanol metabolism.

To characterize metabolic factors potentially associated with alcohol-induced heart disease, myocardial ethanol intermediary metabolism was studied in isolated, perfused rabbit hearts and whole heart homogenates. Results showed that intact rabbit hearts and homogenates of rabbit left ventricle incorporate carbon-14-labeled ethanol at 20 and 59 nmol/g/h, respectively, into a neutral lipid species that co-migrates with triacylglycerides in standard chromatographic solvent systems. After isolation and purification by thin layer chromatography in an apolar solvent system, the labeled species were identified by gas chromatographic-mass spectral analysis to be a family of fatty acid ethyl esters. Heat inactivation of incorporation and the kinetics of formation of products suggest that the process is enzymatic. Gas chromatography identified the fatty acid components as predominantly unsaturated moieties, especially oleic, linoleic, and arachidonic acids. These results provide insight into potential biochemical mechanisms contributing to the triacylglyceride accumulation, decreased beta oxidation of fatty acids, and other lipid abnormalities typical of effects of ethanol on the heart.

Synthesis of various kinds of esters by four microbial lipases.

Ester synthesis by microbial lipases, using homogeneous enzyme preparations, were investigated. The amount of synthesized ester was estimated by alkalimetry, and products were identified by thin-layer chromatography and infrared spectroscopy. Lipases from Aspergillus niger, Rhizopus delemar, Geotrichum candidum and Penicillium cyclopium synthesized esters from oleic acid and various primary alcohols. Only Geotrichum candidum lipase synthesized esters of secondary alcohols. Esters of tertiary alcohols, phenols or sugar alcohols were not synthesized by any lipase. Rather high concentrations of alcohol were required to synthesize the esters of ethylene glycol, propylene glycol or trimethylene glycol. Lipases from Aspergillus niger and Rhizopus delemar synthesized oleyl esters of various fatty acids and some dibasic acids. In contrast, lipases from Geotrichum candidum and Penicillium cyclopium synthesized oleyl esters only from medium or long chain fatty acids.

Differential uptake and metabolism of sitosterol and cholesterol by Achlya, Pythium, and Phytophthora species.

The relative ability of isolates of Achlya bisexualis and A. ambisexualis and isolates of Pythium and Phytophthora to take up and metabolize sitosterol and cholesterol was studied. Species of Pythium and Phytophthora took up cholesterol and sitosterol efficiently, whereas Achlya species took up booth sterols inefficiently. Species of Pythium and Phytophthora produced a polar metabolite and esters from sitosterol as they did from cholesterol. Achlya species did not produce the polar metabolite from either sterol. In these experiments Achlya species produced esters only from cholesterol; however, their failure to produce esters from sitosterol may have been due to the higher sitosterol than cholesterol concentration.

Histochemical changes in gastric mucosubstances in patients with acute and chronic ulcer disease.

The several types of epithelial cells in human gastric mucosa produce different mucosubstances. The surface epithelium largely forms a neutral mucosubstance except that in about two-thirds of the specimens the deep foveolar cells produce a slight to moderate amount of a mucosubstance apparently containing sulfate esters and carboxyl groups. Mucous neck cells often exhibit a neutral mucosubstance but in about onehalf of the stomachs reveal a slight to moderate reactivity indicative of sulfated mucosubstance. Chief cells contain a sulfated mucosubstance with unique histochemical properties. Mast cells vary widely in prevalence but those in the gastric mucosa appear depleted of stored mucosubstances when compared with those in the gastric submucosa or the esophagus. The sulfated mucosubstance normally abundant in human as in canine chief cells appears consistently depleted in patients with stress ulcer or hemorrhagic gastritis. In addition, mucus often appears depleted in the surface epithelium and interstitial edema is present in the superficial mucosa of these patients. These findings appear consistent with the view that biosynthetic activity in chief cells and surface epithelial cells is impaired perhaps secondary to shock-induced circulatory changes in gastric mucosa of patients with stress ulcer or hemorrhagic gastritis.