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%.

Effect of glucose and long-chain fatty acids on synthesis of long-chain alcohols by Candida albicans.

Candida albicans, grown aerobically in glucose-containing media, produced C14, C16 and C18 saturated long-chain alcohols only after the end of exponential growth. Contents of C14 alcohols were always lowest, and C16 and C18 alcohol contents about equal. Contents of all three classes of alcohol increased as the concentration of glucose in aerobic cultures harvested after 168 h incubation was raised from 1.0 to 30.0% (w/v). However, in 168 h anaerobic cultures, greatest long-chain alcohol contents in organisms were obtained using media containing 10% (w/v) glucose. Substituting glucose (10%, w/v) with the same concentration of galactose in aerobic cultures greatly decreased contents of long-chain alcohols, while inclusion of 10% (w/v) glycerol virtually abolished their synthesis. Supplementing anaerobic cultures with odd-chain fatty acids induced synthesis of odd-chain alcohols. Maximum conversion of fatty acid to the corresponding long-chain alcohol was observed with heptadecanoic acid. The effect of glucose on production of heptadecanol from exogenously provided heptadecanoic acid was similar to that observed on synthesis of the three major even-chain alcohols in media lacking a fatty-acid supplement. Cell-free extracts of organisms catalysed in vitro conversion of palmitoyl-CoA to 1-hexadecanol.

Substrate specificity, regiospecificity and stereospecificity of halogenation reactions catalyzed by non-heme-type bromoperoxidase of Corallina pilulifera.

Many organic compounds were found to be substrates for halogenation reactions catalyzed by the non-heme-type bromoperoxidase found in the red alga Corallina pilulifera. Anisole, 1-methoxynaphthalene and thiophene were converted to o and p-bromoanisoles, 1-methoxy-4-bromonaphthalene and 2-bromothiophene respectively. Regiospecificity of the enzymatic bromination of anisole was tested and found to be the same as in the chemical reaction with NaOBr. The enzyme also acted on substituted alkenes such as styrene, cyclohexene, trans-cinnamic acid, trans-cinnamyl alcohol and cis-propenylphosphonic acid, to give the respective bromohydrin compounds or decarboxylated bromo compound. These bromohydrin compounds were always mixtures of stereoisomers. In the light of the above findings together with the previous studies concerning the halogenation mechanism, the bromoperoxidase of C. pilulifera was considered to have no specific restriction site for these substrates.

Production of long-chain alcohols by yeasts.

Fourteen yeast strains from six genera were analysed for the presence of long-chain alcohols. Six strains from three genera contained long-chain alcohols, highest levels being found in Candida albicans. The alcohols were identified and determined by TLC, GLC and GLC-MS. The major long-chain alcohols synthesized by these organisms were saturated, primary alcohols with C14, C16 or C18 chain length. Unsaturated long-chain alcohols were not detected. In all strains that produced long-chain alcohols, the relative proportions were C16 greater than C18 greater than C14. Long-chain alcohol contents were higher in organisms from anaerobically, as compared with aerobically, grown cultures reaching about 650 micrograms (g dry wt organisms)-1 in stationary-phase cultures of C. albicans. In cultures of C. albicans, synthesis of long-chain alcohols occurred only after the end of exponential growth. The alcohols were predominantly present as free alcohols. The fatty-acyl chain-length profile of the triacylglycerol and to a lesser extent the sterol/wax ester fractions from C. albicans reflected that of the long-chain alcohols produced by this yeast.

Prevention of lipid peroxidation by NAD(P)H in rat liver submitochondrial particles.

In rat liver submitochondrial particles both NADH and NADPH inhibit lipid peroxidation induced by cumene hydroperoxide. Concomitantly with the inhibition of lipid peroxidation, NADH and NADPH strongly stimulate the peroxidase activity of rat liver submitochondrial particles. Rotenone slightly prevents both the protective effect on malondialdehyde formation and peroxidase activity. The peroxidase activity of rat liver submitochondrial particles was attributed to the NAD(P)H-mediated reduction of mitochondrial cytochrome P-450 which can act upon hydroperoxides, by decomposing them to alcohols.

Frequency-pulsed electron capture gas-liquid chromatographic analysis of metabolites produced by Clostridium difficile in broth enriched with amino acids.

Clostridium difficile strain CDC A-567 was cultured in Trypticase (BBL Microbiology Systems)-yeast-salt broth supplemented with 0.2% L-leucine, L-norleucine, L-isoleucine, L-tyrosine, or L-tryptophan. Four extractions were done on the spent medium, three at pH 2 and one at pH 10, using CHCL3 or ether. Derivatizations were done with trichloroethanol, heptafluorobutyric anhydride, and heptafluorobutyric anhydride-ethanol. All samples were analyzed with frequency-pulsed electron capture gas-liquid chromatography. A dedicated computer was used to assist in data analysis. C. difficile produced both short-chain and aromatic acids in Trypticase-yeast-salt broth; hydroxy acids were also detected. p-Cresol, indoleacetic acid, 4-methylthio-2-hydroxybutyric acid, and some unidentified alcohols were observed. The basic chloroform extraction contained cadaverine and putrescine. Leucine, norleucine, and isoleucine influenced the production of C5 and C6 acids and alcohols. L-Tyrosine underwent successive degradation to produce p-cresol and aromatic acids as final products. Tryptophan increased the production of indoleacetic, indolepropionic, and indolebutyric acids. Isocaproic acid was produced in relatively high concentrations regardless of medium substitution. The consistent production of iC6 under various substrate conditions indicates that the production of this compound might be consistent enough in vitro to form the basis of a rapid test for detection of C. difficile in stool specimens by frequency-pulsed electron capture gas-liquid chromatography.

Identification of volatile organic compounds produced by fluorescent pseudomonads on chicken breast muscle.

Four different fluorescent pseudomonads were isolated from spoiled, uncooked chicken breasts and were grown in pure culture on initially sterile chicken breast muscle at 2 to 6 degrees C for 14 days. The volatile compounds produced by each culture were concentrated on a porous polymer precolumn and separated and identified by high-resolution gas chromatography-mass spectrometry.

Headspace analysis of volatile metabolites of Pseudomonas aeruginosa and related species by gas chromatography-mass spectrometry.

Gas chromatographic-mass spectrometric analysis of headspace volatiles was performed on cultures of 11 strains of Pseudomonas aeruginosa and 1 strain each of Pseudomonas cepacia, Pseudomonas putida, Pseudomonas putrefaciens, Pseudomonas fluorescens, and Pseudomonas maltophilia. All strains of Pseudomonas aeruginosa produced a distinctive series of odd-carbon methyl ketones, particularly 2-nonanone and 2-undecanone, and 2-aminoacetophenone. The other strains failed to produce 2-aminoacetophenone. Two sulfur compounds, dimethyldisulfide and dimethyltrisulfide, were present in strains of P. aeruginosa and in variable amounts in other species. Butanol, 2-butanone, 1-undecene, and isopentanol were also detected in P. aeruginosa cultures.

[Formation of higher alcohols by Saccharomyces carlsbergensis from branched-chain amino acids and their keto analogs]. / Obrazovanie vysshikh spirtov Saccharomyces carlsbergensis iz razvetvlennykh aminokislot i ikh ketoanalogov.

A new method of chemical synthesis of alpha-ketoisocaproic acid (the keto analogue of L-leucine) is described. It has been shown that the resting cells of Saccharomyces carlsbergensis 776, in the stationary state of biomass, produce mainly higher alcohols: isobutanol from L-valine and its keto analogue; optically active amylol only from L-isoleucine and its keto analogue; isoamylol only from L-leucine and its keto analogue. "Nonspecific" formation of n-propanol from L-valine, L-isoleucine and their keto analogues, as well as that of isobutanol from L-isoleucine and its keto analogue, has been also found at pH 7.0. Formation of higher alcohols from alpha-keto acids has an acidic pH optimum while that from L-amino acids has a neutral or a weakly alkaline pH optimum. Formation of isobutanol from L-valine is an exception. The dependence of higher alcohol formation on the pH and the kinetics of their accumulation suggest that higher alcohols are produced from L-amino acids in at least three sequential reactions: transamination, decarboxylation of the keto analogue being formed, and reduction of the aldehyde; formation of higher alcohols from alpha-keto acids involves two reactions: decarboxylation and reduction. Transamination and decarboxylation are limiting steps in the process in the former case, and decarboxylation in the latter.

Electron capture gas chromatography study of the acid and alcohol products of Clostridium septicum and Clostridium chauvoei.

The metabolic products produced by several strains of Clostridium septicum obtained from patients and animals, along with strains of Clostridium chauvoei, were studied in chopped meat glucose medium by electron capture gas-liquid chromatography (EC-GLC). The strains of C. septicum and C. chauvoei were shown to comprise five different metabolic groups. Both the EC-GLC study and the O and H antigenic study performed previously showed that strains of C. septicum comprise a heterogeneous group. One type of metabolic profile was found only in strains of C. chauvoei. The O antigen types and EC-GLC metabolic types of C. septicum correlated fairly well in isolates from cancer patients but not in stock culture and animal isolates.

[Effect of furfural on the energy metabolism of fodder yeasts]. / Vliianie furfurola na énergeticheskii obmen kormovykh drozhzhei

The effect of furfurol on the rate of oxygen uptake and carbon dioxide production by Candida tropicalis cells was studied. Furfurol was shown to inhibit significantly the yeast respiration, inducing aerobic glycolysis. Enhanced carbon dioxide production in the presence of furfurol was followed by an intensive degradation of trehalose and ethanol accumulation in the medium.

[Formation of primary alcohols and palmitic acid in the microbiological oxidation of hexadecane]. / Obrazovanie pervichnykh spirtov i pal'mitinovoi kisloty pri mikrobiologicheskom okislenii geksadekana

Forty seven microbial strains oxidized hexadecane to form primary cetol and palmitic acid. The maximum quantitiy of the compounds was accumulated in the exponential phase of growing cultures (for 72 hours) and for 6 to 16 hours of incubation of resting cell suspensions. Candida yeast were shown by the gas-liquid chromatography method to be the most active producers of cetol.

Intermediates in the metabolism of m-carboxy-substituted aromatic amino acids in plants. Phenylpyruvic acids, mandelic acids, and phenylglyoxylic acids.

Tracer experiments with 14C-labelled precursors in Iris times hollandica cv. Wedgwood, Reseda Iutea L. And Keseda Odorata L. have demonstrated that 3-(3-carboxyphenyl) alanine and 3-(3-carboxy-4-hydroxyphenyl) alanine can be derived from the corresponding pyruvic acids, presumably by unspecific transaminations, and that (3-carboxyphenyl) glycine and (3-carboxy-4-hydroxyphenyl) glycine can be derived from the corresponding phenylglyoxylic acids. The glycine derivatives are derived from the alanine derivatives, and the corresponding mandelic acids are intermediates in these transformations. The corresponding phenylacetic acids are incorporated only slightly into the glycine derivatives, indicating that oxidation at the benzylic position in the C6-C3 compounds takes place early in the transformation. The corresponding cinamic acids are not metabolized at all in the plants.

Identification of major high-boiling volatile compounds produced during refrigerated storage of haddock fillets.

The two major high-boiling volatile compounds produced during refrigerated storage of haddock fillets were found by gas chromatography and mass spectroscopy to be phenethyl alcohol and phenol.

Taxonomic significance of phenethyl alcohol production by Achromobacter isolates from fishery sources.

The formation of phenethyl alcohol from L-phenylalanine and ethanol by achromobacter isolates of fishery origin was found to be taxonomically significant for such organisms. Phenylpyruvate, the direct oxidative deamination product of L-phenylalanine, was found to serve as an intermediate precursor to phenethyl alcohol formation. Among ten Acinetobacter isolates examined, none produced phenethyl alcohol. Among nine Moraxella isolates examined, one produced phenethyl alcohol.

Involvement of vitamin B6 in the dethiomethylation of methionine by rumen microorganisms.

The dethiomethylation of methionine by a dialyzed extract obtained from the protozoa-rich fraction of rumen fluid is stimulated 2.5-fold by pyridoxal phosphate and strongly inhibited by deoxypyridoxine, a pyridoxal phosphate antagonist. These effects are not seen with undialyzed extracts or with whole rumen fluid. It is suggested that the anaerobic dethiomethylation of methionine by rumen microorganisms requires pyridoxal phosphate as a cofactor.

Microbial water relations: features of the intracellular composition of sugar-tolerant yeasts.

Several factors contributed to differences in intracellular composition between sugar-tolerant (osmophilic) and nontolerant species of yeast. One such factor was the difference in accumulation of those nonelectrolytes whose uptake was not dominated by vigorous metabolism. In such cases (lactose and glycerol), the sugar-tolerant species had a much lower capacity for the solute than did the nontolerant species. Sucrose uptake was consistently different between all sugar-tolerant strains on the one hand and all nontolerant strains on the other. The difference was attributable in part to metabolism of sucrose by the nontolerant yeasts. The major difference between the two types of yeast, however, was the presence of one or more polyhydric alcohols at high concentrations within each of the sugar-tolerant strains but none of the nontolerant strains. In most cases the major polyol was arabitol. The solute concentration (and, hence, water availability) of the growth medium affected both the amount of arabitol produced by Saccharomyces rouxii and the proportion retained by the yeast after brief washing with water at 0 C. When the yeast was suspended in a buffer at 30 C, the polyol leaked out at a slow, constant, reproducible rate. The polyene antibiotic amphotericin B caused rapid release of polyol by the yeast, the rate being proportional to amphotericin concentration. Contact of the yeast with glucose (1 mM) caused an extremely rapid ejection of polyol which lasted less than 40 s. Some implications of these results are discussed, as is the role of the polyol as a compatible solute in determining the water relations of the yeast.