The inhibition of phospholipid synthesis in escherichia coli by phenethyl alcohol.

The kinetics of lipid metabolism during phenethyl alcohol treatment of Escherichia coli were examined. Phenethyl alcohol at a non-bacteriostatic concentration reduces the accumulation of [32-P] phosphate into phospholipids and alters the phospholipid composition of the cell membrane. The changes in phospholipid composition are a result of the inhibitory effect of phenethyl alcohol on the rates of synthesis of the individual phospholipids. The inhibition in the rate of phosphatidylethanolamine synthesis by phenethyl alcohol was twice the inhibition in the rate of phosphatidyglycerol synthesis. The de novo rate of cardiolipin synthesis was only slightly inhibited. However, net cardiolipin accumulation increased during phenethyl alcohol treatment due to a more rapid turnover of phosphatidylglycerol to cardiolipin. Phenethyl alcohol also altered the fatty acid composition of the cell as a result of its inhibitory effect on the rate of individual fatty acid synthesis. However, the inhibition of phospholipid synthesis was not reversed by fatty acid supplementation of phenethyl alcohol treated cells. This result indicates that phenethyl alcohol does not inhibit phospholipid synthesis solely at the level of fatty acid synthesis.     

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.     

Modification of membrane lipids. Phenethyl alcohol-induced alteration of lipid composition in Tetrahymena membranes.

Tetrahymena pyriformis NT-I cells in the early-logarithmic phase were incubated with phenethyl alcohol (2-phenylethanol) and effects on the lipid composition were examined in various membranes. 1. There was a marked modification in phospholipid head, as well as fatty acyl group composition in pellicles, mitochondria and microsomes of the phenethyl alcohol-treated cells. Compared with membranes of the control cells, the membranes from phenethyl alcohol-treated cells were found to contain a higher level of phosphatidylcholine content with the compensating decrease in phosphatidylethanolamine, while 2-aminoethylphosphonolipid showed only a slight decrease in these membranes. The acyl group profile of membrane phospholipids in the presence of phenethyl alcohol was also modified so that a profound elevation of the content of polyunsaturated fatty acids, linoleic and gamma-linolenic acids. The major monounsaturate, palmitoleate decreased. Such lipid alteration is a reversible process, and therefore upon removal of phenethyl alcohol the modified lipid composition returned to normal. 2. By freeze-fracture electron microscopy in combination with temperature quenching, the outer alveolar membrane of the phenethyl alcohol-treated cell was observed to reveal less aggregation of intercalated-membrane particles, as compared with the control membrane. The quantitative analysis of the thermotropic lateral movement of membrane particles provided evidence that the membrane in the phenethyl alcohol-treated cell became more fluid. Such fluidizing effects may result from an increase in the acyl group unsaturation and also in the phosphatidylcholine content. 3. With regard to the mechanism responsible for the marked decrease in palmitoleate in membrane phospholipids, there was found a depressed conversion of the palmitate to palmitoleate in the phenethyl alcohol-treated cells. It was further suggested that the drug may have an inhibitory effect on the synthesis of palmitoyl-CoA desaturase involving the (16 : 0 leads to 16 : 1) conversion. Also, it was demonstrated that the increase in a precursor-product fashion of phosphatidylcholine with the corresponding decrease in phosphatidylethanolamine was not due to transformation of phosphatidylethanolamine to phosphatidylcholine through stepwise methylation.     

Patterns of Protein Production in Myxococcus xanthus During Spore Formation Induced by Glycerol, Dimethyl Sulfoxide, and Phenethyl Alcohol

Spore formation of Myxococcus xanthus can occur not only on agar plates during fruiting body formation, but also in a liquid culture by simply adding glycerol, dimethyl sulfoxide, or phenethyl alcohol to the culture. This chemically-induced spore formation occurs synchronously and much faster than that occurring during fruiting body formation. Dramatic changes in patterns of protein synthesis were observed during chemically-induced spore formation, as had previously been observed during fruiting body formation (Inouye et al., Dev. Biol. 68:579-591, 1979). However, the production of protein S, one of the major development-specific proteins during fruiting body formation, was not detected at all, although protein U, another development-specific protein, was produced in a late stage of spore formation as in the case of fruiting body formation. This indicates that the control of the gene expression during chemically-induced spore formation is significantly different from that during fruiting body formation. It was also found that during spore formation, every cell seems to have a potential to form a spore regardless of its age, since smaller cells as well as larger cells separated by sucrose density gradient centrifugation could equally form spores upon the addition of glycerol. Patterns of protein synthesis were almost identical for all the three chemicals. However, the final yield of spores was significantly different depending upon the chemicals used. When phenethyl alcohol was added with glycerol or dimethyl sulfoxide, the final yields were determined by the multiple effect of the two chemicals added. This suggests that although these chemicals are able to induce the gene functions required for spore formation, they may have inhibitory effects on some of the gene functions or the processes of spore formation.     

Action of phyenethyl alcohol on the synthesis of macromolecules in Escherichia coli

Prevost, C. (University of California, Berkeley), and V. Moses. Action of phenethyl alcohol on the synthesis of macromolecules in Escherichia coli. J. Bacteriol. 91:1446-1452. 1966.-A kinetic study of the effects of various concentrations of phenethyl alcohol on the synthesis of ribonucleic acid (RNA), deoxyribonucleic acid (DNA), protein, and beta-galactosidase in Escherichia coli has confirmed that RNA synthesis, rather than DNA synthesis, is first and most affected by phenethyl alcohol. The presence of inducer did not protect beta-galactosidase synthesis from inhibition by phenethyl alcohol. Little preferential inhibition of beta-galactosidase synthesis was observed; this is in contrast to the severe catabolite repression which results from partial inhibition of total protein synthesis caused by chloramphenicol or starvation for a required amino acid. We found no evidence that messenger RNA synthesis was inhibited to a greater extent than total RNA synthesis.     

Novel scheme for biosynthesis of aryl metabolites from L-phenylalanine in the fungus Bjerkandera adusta

Aryl metabolite biosynthesis was studied in the white rot fungus Bjerkandera adusta cultivated in a liquid medium supplemented with L-phenylalanine. Aromatic compounds were analyzed by gas chromatography-mass spectrometry following addition of labelled precursors ((14)C- and (13)C-labelled L-phenylalanine), which did not interfere with fungal metabolism. The major aromatic compounds identified were benzyl alcohol, benzaldehyde (bitter almond aroma), and benzoic acid. Hydroxy- and methoxybenzylic compounds (alcohols, aldehydes, and acids) were also found in fungal cultures. Intracellular enzymatic activities (phenylalanine ammonia lyase, aryl-alcohol oxidase, aryl-alcohol dehydrogenase, aryl-aldehyde dehydrogenase, lignin peroxidase) and extracellular enzymatic activities (aryl-alcohol oxidase, lignin peroxidase), as well as aromatic compounds, were detected in B. adusta cultures. Metabolite formation required de novo protein biosynthesis. Our results show that L-phenylalanine was deaminated to trans-cinnamic acid by a phenylalanine ammonia lyase and trans-cinnamic acid was in turn converted to aromatic acids (phenylpyruvic, phenylacetic, mandelic, and benzoylformic acids); benzaldehyde was a metabolic intermediate. These acids were transformed into benzaldehyde, benzyl alcohol, and benzoic acid. Our findings support the hypothesis that all of these compounds are intermediates in the biosynthetic pathway from L-phenylalanine to aryl metabolites. Additionally, trans-cinnamic acid can also be transformed via beta-oxidation to benzoic acid. This was confirmed by the presence of acetophenone as a beta-oxidation degradation intermediate. To our knowledge, this is the first time that a beta-oxidation sequence leading to benzoic acid synthesis has been found in a white rot fungus. A novel metabolic scheme for biosynthesis of aryl metabolites from L-phenylalanine is proposed.     

Inhibition of excision repair of DNA in u.v.-irradiated Escherichia coli by phenethyl alcohol

Membrane-specific drugs such as procaine and chlorpromazine have been shown to inhibit excision repair of DNA in u.v.-irradiated E. coli. One possible mechanism is that, if association of DNA with the cell membrane is essential for excision repair, this process may be susceptible to drugs affecting the structure of cell membranes. We examined the effect of phenethyl alcohol, which is a membrane-specific drug and known to dissociate the DNA-membrane complex, on excision repair of DNA in u.v.-irradiated E. coli cells. The cells were irradiated with u.v. light and then held at 30 degrees C in buffer (liquid-holding) in the presence or absence of phenethyl alcohol. It was found that phenethyl alcohol inhibits the liquid-holding recovery in both wild-type and recA strains, corresponding to its dissociating action on the DNA-membrane complex. Thus, the association of DNA with cell membrane is an important factor for excision repair in E. coli. Procaine did not show the dissociating effect, suggesting that at least two different mechanisms are responsible for the involvement of cell membrane in excision repair of DNA in E. coli.     

Production of aryl metabolites in solid-state fermentations of the white-rot fungus Bjerkandera adusta

Bjerkandera adusta produced aromatic compounds such as benzaldehyde (bitter almond aroma), benzyl alcohol and benzoic acid from L-phenylalanine (3 g kg^–1). Two supports for the fungus, wheat bran (organic support) and Perlite (mineral support), gave optimal production with water contents of 66% and 60%, respectively. Benzyl alcohol (4.53 g kg^–1) and benzaldehyde (1.56 g kg^–1) were produced after 4 days on wheat bran respectively with 20 and 30 g L-phenylalanine kg^–1. Aryl alcohol oxidase activity, which oxidises benzyl alcohol to benzaldehyde, was only detected when the fungus was grown on wheat bran, the support which promotes the most benzaldehyde production. Results are compared with those obtained in submerged liquid cultures.     

Genetic Determination of Resistance to Acriflavine, Phenethyl Alcohol, and Sodium Dodecyl Sulfate in Escherichia coli

Wild-type strains of Escherichia coli K-12 are resistant to acriflavine. Gene acrA(+) which determines resistance to acriflavine is located near the lac region of the chromosome. This gene determines not only resistance to basic dyes but also resistance to phenethyl alcohol. Acriflavine resistance was transmitted, together with phenethyl alcohol resistance, from a resistant Hfr strain to a sensitive recipient by mating. Reversion of the mutant gene acrA1 (phenotypically acriflavine-sensitive) to acriflavine resistance was accompanied by a change from phenethyl alcohol sensitivity to resistance, and conversely the revertants selected for phenethyl alcohol resistance were resistant to acriflavine. A suppressor mutation, sup-100, closely linked to the acr locus, suppresses the acrA1 gene (phenotypically acriflavine-resistant), but does not determine resistance to phenethyl alcohol and basic dyes other than acriflavine. The genetic change in the locus acrA1 to types resistant to basic dyes and phenethyl alcohol was accompanied by an increase in resistance to sodium dodecyl sulfate, a potent solvent of lipopolysaccharide and lipoprotein. It is suggested that gene acrA determines synthesis of a membrane substance. The system seemed to be affected strongly by the presence of inorganic phosphate.     

Pretreatment of hamster cells with phenethyl alcohol alters cell surface glycoproteins and inhibits vesicular stomatitis virus growth.

Chinese hamster ovary cells cultured in the presence of phenethyl alcohol exhibit obvious changes in cell surface galactose and galactosamine glycoproteins as determined by the galactose-oxidase[3H]borohydride technique and SDS gel electrophoresis. Cells pretreated with phenethyl alcohol (drug was removed before infection) were not as effective as hosts for vesicular stomatitis virus as untreated cultures. A minimum pretreatment time with 0.1% phenethyl alcohol of about 8 h was required before a reduction in virus growth was observed. It is proposed that phenethyl alcohol pretreatment as outlined in this report leads to a modification of the host cellular membrane resulting in the inhibition of virus replication.     

The effect of phenethyl alcohol on in vitro DNA synthesis in Escherichia coli.

The effect of phenethyl alcohol on DNA synthesis was examined using several in vitro systems of Escherichia coli H560; i.e., ether-treated cells, membrane fractions and folded chromosomes fortified with DNA polymerase. In all systems, the incorporation of deoxyribonucleotides was much reduced for the phenethyl alcohol-treated cells compared with the non-treated cells. The total activity of DNA polymerases in polA1 cells (mostly DNA polymerase II) was not impaired for the phenethyl alcohol-treated cells and the reduction of the rate of DNA synthesis in vitro was ascribed to the reduction of the chromosomal template activity which was related to trypsin sensitive protein components. The analysis of chromosomes from the phenethyl alcohol-treated cells revealed the remarkable reduction of a protein component of molecular weight approx. 58 000 in contrast with a protein component of molecular weight approx. 30 000.     

Controlled formation of volatile components in cider making using a combination of Saccharomyces cerevisiae and Hanseniaspora valbyensis yeast species

The effect of pure and mixed fermentation by Saccharomyces cerevisiae and Hanseniaspora valbyensis on the formation of major volatile components in cider was investigated. When the interaction between yeast strains of S. cerevisiae and H. valbyensis was studied, it was found that the two strains each affected the cell growth of the other upon inoculation of S. cerevisiae during growth of H. valbyensis. The effects of pure and mixed cultures of S. cerevisiae and H. valbyensis on alcohol fermentation and major volatile compound formation in cider were assessed. S. cerevisiae showed a conversion of sugar to alcohol of 11.5%, while H. valbyensis produced alcohol with a conversion not exceeding 6%. Higher concentrations of ethyl acetate and phenethyl acetate were obtained with H. valbyensis, and higher concentrations of isoamyl alcohol and isobutyl were formed by S. cerevisiae. Consequently, a combination of these two yeast species in sequential fermentation was used to increase the concentration of ethyl esters by 7.41–20.96%, and to decrease the alcohol concentration by 25.06–51.38%. Efficient control of the formation of volatile compounds was achieved by adjusting the inoculation time of the two yeasts.     

A model of closely assembled consecutive enzymes on membranes: formation of hydroxycinnamic acids from L-phenylalanine on thylakoids of Dunaliella marina.

p-Hydroxycinnamic acid was found to be located within the plastids of the green alga Dunaliella marina. Thylakoid fractions desintegrated by ultrasonic treatment were capable of converting L-phenylalanine into o- and p-hydroxycinnamic acids; the hydroxylation reaction was increased by addition of NADPH. Hydroxycinnamic acids produced when [3-14C]cinnamate was incubated with varying amounts of [4'-3H]L-phenylalanine exhibited a 3H/14C ratio 10-150 times higher than that of the cinnamic acid reisolated from the incubation mixture. The lack of equilibration between cinnamate formed from L-phenylalanine and cinnamate added to the solution supports the hypothesis that cinnamate as an intermediate in hydroxycinnamate formation remains bound to the membrane enzyme complex. A model of membrane-bound multienzyme complexes is proposed for the conversion of aromatic amino acids into phenols.     

Impact of biocides on biofilm formation by methicillin-resistant Staphylococcus aureus (ST239-SCCmecIII) isolates

Procedures of sterilization and disinfection are essential to ensure that medical and surgical instruments will not transmit infectious pathogens to patients. In the present paper, we tested the residual effect of these compounds on biofilm formation and its efficiency in disrupting preformed biofilms using methicillin-resistant Staphylococcus aureus (MRSA) isolates of the lineage ST239-SCCmecIII. All compounds examined, except 70% alcohol, caused a significant impairment in biofilm formation with concomitant inhibition of cell growth. Among the compounds examined, 10% povidone-iodine (PVP-I) was the only antiseptic that exhibited more than 90% reduction of both biofilm formation and dispersion. In the group of sterilants and disinfectants, a formulation containing 7% hydrogen peroxide and 0.2% peracetic acid (HP-PA), and sodium hypochlorite with 1% active chlorine (NaOCl) were equally effective.     

Diurnal changes in the concentrations of 2-phenylethyl β-D-glucopyranoside and the corresponding volatile aglycone in the tissue and headspace of Trifolium repens L. florets

Distinct synchronous diurnal rhythms were detected in the concentrations of phenethyl alcohol and phenethyl acetate in the tissue of blooming Trifolium repens florets. Corresponding rhythmic oscillations were observed for the same two compounds in the floral headspace. Maximum content of the volatiles in the tissue and headspace was observed 3–9 h after initiation of the photophase. The concentrations of phenethyl alcohol and phenethyl acetate in the tissue increased significantly during floral development. At full bloom the tissue contained amounts sufficient to support 2–3 h of emission. Several observations suggested that esterification of phenethyl alcohol was the source for phenethyl acetate. Trimethylsilyl derivatization and enzymatic hydrolysis of fractionated flower extracts identified 2‐phenylethyl β‐d ‐glucopyranoside as the major glucoside in the florets. The pool of glucosides increased significantly during floral development and at full bloom 97% of the phenethyl alcohol was bound as glucoside. The concentration of 2‐phenylethyl β‐d ‐glucopyranoside did not vary in a rhythmic diurnal manner. The dynamics among the diurnal rhythmic phenomena in the tissue and headspace and the fraction of volatiles bound as glucosides is discussed.     

Phenethyl Alcohol as a Suppressor of the EnvA Phenotype Associated with the envA Gene in Escherichia coli K-12

In Escherichia coli K-12 the envA gene was previously shown to mediate chain formation and a decreased tolerance to several antibacterial agents. Phenethyl alcohol at low concentrations has now been found to increase the tolerance to actinomycin D, ampicillin, rifampin, and gentian violet in strains containing envA. The increased tolerance to gentian violet was correlated to a decreased uptake of the dye. A phenotype suppression of chain formation and colony morphology in envA mutants was also obtained. Except for an increase in palmitic acid, chemical analysis revealed no differences between an envA and its wild-type strain in the lipopolysaccharide part of the envelope. However, a decrease in the amount of phosphatidylglycerol and a C18: 1 fatty acid was observed in the extractable lipids of a strain containing envA. Growth in the presence of phenethyl alcohol reversed the changes in fatty acid and the phospholipid composition. Phenethyl alcohol was found to cause an immediate but transient inhibition of ribonucleic acid synthesis. It is suggested that this inhibition affects the penetrability barrier of the outer cell envelope layers in strains containing envA.     

Volatile Flavor Compounds Produced by Molds of Aspergillus, Penicillium, and Fungi imperfecti

Strains of molds Aspergillus niger, A. ochraceus, A. oryzae, A. parasiticus, Penicillium chrysogenum, P. citrinum, P. funiculosum, P. raistrickii, P. viridicatum, Alternaria, Cephalosporium, and Fusarium sp. were grown on sterile coarse wheat meal at 26 to 28 C for 120 h. The volatiles from mature cultures were distilled at low temperature under reduced pressure. The distillates from traps -40 and -78 C were extracted with methylene chloride and subsequently concentrated. All the concentrates thus obtained were analyzed by gas-liquid chromatography, mass spectrometry, chemical reactions of functional groups, and olfactory evaluation. Six components detected in the culture distillates were identified positively: 3-methylbutanol, 3-octanone, 3-octanol, 1-octen-3-ol, 1-octanol, and 2-octen-1-ol. They represented 67 to 97% of all the volatiles occurring in the concentrated distillate. The following 14 components were identified tentatively: octane, isobutyl alcohol, butyl alcohol, butyl acetate, amyl acetate, octyl acetate, pyridine, hexanol, nonanone, dimethylpyrazine, tetramethylpyrazine, benzaldehyde, propylbenzene, and phenethyl alcohol. Among the volatiles produced by molds, 1-octen-3-ol yielding a characteristic fungal odor was found predominant.     

Induction of alkaline phosphatase in Escherichia coli. Effect of phenethyl alcohol.

Induction of alkaline phosphatase, an enzyme located in the periplasmic region of Escherichia coli, was inhibited by phenethyl alcohol, an agent believed to alter the cell membrane structure. Studies to elucidate mechanism of this inhibition showed that while phenethyl alcohol arrested the incorporation of [3H]leucine into active alkaline phosphatase, it did allow substantial incorporation of the label into inactive monomer subunits of the enzyme. These results suggest that phenethyl alcohol may not interfere with the de novo synthesis of monomer subunits of the enzyme but arrest conversion of these into active dimer enzyme presumably by its primary action on the cell membrane structure.     

Recovery of Impaired Cell Division of UV-Irradiated Escherichia coli by Several Chemical Agents Affecting Cell Membrane

We examined the effect of various agents on the cell division of E. coli B(Sm^r) irradiated with ultraviolet (UV) light. It was found that the impaired cell division was reversed by one of various agents such as higher fatty acids, lower alcohols, terpineol, phenethyl alcohol, streptomycin, ribonuclease and EDTA. These agents, except RNase, showed the maximum activity of recovery just below a concentration causing a complete suppression of cell growth.     

女贞小蜡树的酚性配糖体成分研究

从女贞小蜡树 (LigustrumsinenseLour.)茎叶甲醇提取物的水溶性部分得到 1个新的和 6个已知酚性配糖体成分。它们是两个已知的裂环环烯醚萜类化合物 :1 0 hydroxyoleuropein( 1 )和specneuzhenide( 2 ) ,五个苯乙醇类化合物 :3 ,4 二羟基苯乙醇 ( 3 ) ,3 ,4 二羟基苯乙醇 2′ O β D 吡喃葡萄糖甙 ( 4 ) ,3 甲氧基 苯乙醇 4 O β D 吡喃葡萄糖甙 ( 5 ) ,4 羟基苯乙醇 ( 6) ,4 羟基苯乙醇 2′ O β D 吡喃葡萄糖甙 ( 7) ,化合物 5为新化合物 ,命名为小蜡甙A(sinenosideA) ,经理化和波谱分析鉴定了论文中的所有化合物的结构