Higher alcohols are one of the main by-products of Saccharomyces cerevisiae in brewing. High concentration of higher alcohols in alcoholic beverages easily causes headache, thirst and other symptoms after drinking. It is also the main reason for chronic drunkenness and difficulty in sobering up after intoxication. The main objective of this review is to present an overview of the flavor characteristics and metabolic pathways of higher alcohols as well as the application of mutagenesis breeding techniques in the regulation of higher alcohol metabolism in S. cerevisiae. In particular, we review the application of metabolic engineering technology in genetic modification of amino transferase, α-keto acid metabolism, acetate metabolism and carbon-nitrogen metabolism. Moreover, key challenges and future perspectives of realizing optimization of higher alcohols metabolism are discussed. This review is intended to provide a comprehensive understanding of metabolic regulation system of higher alcohols in S. cerevisiae and to provide insights into the rational development of the excellent industrial S. cerevisiae strains producing higher alcohols.
Alcoholic Beverages ; Alcohols/analysis* ; Fermentation ; Saccharomyces cerevisiae/metabolism* ; Saccharomyces cerevisiae Proteins/metabolism*

OBJECTIVE: To explore the role of mitochondrial permeability transition pore (mPTP) in mediating the protective effect of gastrodin against oxidative stress damage in H9c2 cardiac myocytes. METHODS: H9c2 cardiac myocytes were treated with HO, gastrodin, gastrodin+HO, cyclosporin A (CsA), or CsA+gas+HO group. MTT assay was used to detect the survival ratio of H9c2 cells, and flow cytometry with Annexin V-FITC/PI double staining was used to analyze the early apoptosis rate after the treatments. The concentration of ATP and level of reactive oxygen species (ROS) in the cells were detected using commercial kits. The mitochondrial membrane potential of the cells was detected with laser confocal microscopy. The expression of cytochrome C was detected with Western blotting, and the activity of caspase-3 was also assessed in the cells. RESULTS: Gastrodin pretreatment could prevent oxidative stress-induced reduction of mitochondrial membrane potential, and this effect was inhibited by the application of CsA. Gastrodin significantly lowered the levels of ROS and apoptosis-related factors in HO-exposed cells, and such effects were reversed by CsA. CsA significantly antagonized the protective effect of gastrodin against apoptosis in HO-exposed cells. CONCLUSIONS Gastrodin prevents oxidative stress-induced injury in H9c2 cells by inhibiting mPTP opening to reduce the cell apoptosis.
Adenosine Triphosphate ; analysis ; Apoptosis ; drug effects ; Benzyl Alcohols ; antagonists & inhibitors ; pharmacology ; Caspase 3 ; analysis ; Cell Line ; Cell Survival ; drug effects ; Cyclosporine ; pharmacology ; Cytochromes c ; analysis ; Glucosides ; antagonists & inhibitors ; pharmacology ; Humans ; Hydrogen Peroxide ; antagonists & inhibitors ; pharmacology ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondrial Membrane Transport Proteins ; physiology ; Myocytes, Cardiac ; drug effects ; metabolism ; Oxidative Stress ; Reactive Oxygen Species ; analysis

BACKGROUND: Alcohol is known to affect two epigenetic phenomena, DNA methylation and DNA hydroxymethylation, and iron is a cofactor of ten-eleven translocation (TET) enzymes that catalyze the conversion from methylcytosine to hydroxymethylcytosine. In the present study we aimed to determine the effects of alcohol on DNA hydroxymethylation and further effects of iron on alcohol associated epigenetic changes. METHODS: Twenty-four male Sprague-Dawley rats were fed either Lieber-DeCarli alcohol diet (36% calories from ethanol) or Lieber-DeCarli control diet along with or without iron supplementation (0.6% carbonyl iron) for 8 weeks. Hepatic non-heme iron concentrations were measured by colorimetric assays. Protein levels of hepatic ferritin and transferrin receptor were determined by Western blotting. Methylcytosine, hydroxymethylcytosine and unmodified cytosine in DNA were simultaneously measured by liquid chromatography/mass spectrometry method. RESULTS: Iron supplementation significantly increased hepatic non-heme iron contents (P < 0.05) but alcohol alone did not. However, both alcohol and iron significantly increased hepatic ferritin levels and decreased hepatic transferrin receptor levels (P < 0.05). Alcohol reduced hepatic DNA hydroxymethylation (0.21% ± 0.04% vs. 0.33% ± 0.04%, P = 0.01) compared to control, while iron supplementation to alcohol diet did not change DNA hydroxymethylation. There was no significant difference in methylcytosine levels, while unmodified cytosine levels were significantly increased in alcohol-fed groups compared to control (95.61% ± 0.08% vs. 95.26% ± 0.12%, P = 0.03), suggesting that alcohol further increases the conversion from hydroxymethylcytosine to unmodified cytosine. CONCLUSIONS: Chronic alcohol consumption alters global DNA hydroxymethylation in the liver but iron supplementation reverses the epigenetic effect of alcohol.
Alcohol Drinking* ; Alcohols ; Animals ; Blotting, Western ; Cytosine ; Diet ; DNA Methylation ; DNA* ; Epigenomics ; Ferritins ; Humans ; Iron* ; Liver ; Male ; Methods ; Rats* ; Rats, Sprague-Dawley ; Receptors, Transferrin ; Spectrum Analysis

To establish a HPLC method for simultaneously determining plasma concentrations of gastrodin (Gas) and its metabolites hydroxybenzyl alcohol (HBA), puerarin (Pur) and internal standard (IS) p-hydroxyphenylethanol (Tyr) in rats and studying the pharmacokinetic process and interactions of gastrodin and puerarin after single and combined intravenous injection and oral administration. With Tyr as the internal standard, plasma samples were processed with methanol for protein precipitation, supernatant was dried with N2, and residues were re-dissolved with acetonitrile-0.05% phosphoric acid (20: 80). Chromatography was carried out on an Agilent ZORBAX SB-Aq C18 column (4.6 mm x 250 mm, 5 μm), with acetonitrile-0.05% phosphoric acid as the gradient mobile phase for the gradient elution. The UV detector wavelength was set at 221 nm for Gas HBA and IS and 250 nm for Pur. After the single or combined administration of Gas and Pur, their plasma concentrations in rats were detected. WinNonlin 5.2 pharmacokinetic software and SPSS 17. 0 software were used to respectively calculate pharmacokinetic parameters of each group, make a statistical analysis and compare the pharmacokinetic processes of Gas and Pur after the single or combined administration. According to the results, the absolute recoveries between low, media and high concentrations of Gas, HBA and Pur and IS as well as Tyr were more than 77.20%, with a good linearity (r > 0.999 6, n = 5) for Gas, HBA and Pur within concentration ranges of 0.10-101, 0.03-7.58 and 0.05-5.98 mg xL ('1) respectively. The lower limits of quantification for Gas, HBA and Pur were 0.10, 0.03, 0.05 mg x L(-1), respectively. Their in-ra-day and inter-day precisions were less than 12% with the accuracy between 85. 1% -1 10. %. All of the three substances and IS were stable during the whole analysis process. The findings showed significant differences in the main in vivo pharmacokinetic parame-ers in rats (AUC, C.(max) T,½ T.(max) MRT) after the single and combined administration of Gas and Pur. Either after the oral adminis-ration or after the intravenous injection, parameters showed a lower clearance rate ( L) longer mean residence time ( RT) and higher relative bioavailability, especially after the oral administration. Specifically, the relative bioavailability of the combined oral ad-inistration of Pur was 10. 7 times of that of the single administration, while that of Gas was 1. times of that of the single administra-ion. The combined administration of Gas and Pur can promote the absorption, decrease the elimination rate and prolong the mean resi-ence time. The method is simple and accurate and can be applied in the simultaneous determination of plasma concentrations of Gas, HBA and Pur in rats and the pharmacokinetic studies.
Administration, Oral ; Animals ; Benzyl Alcohols ; administration & dosage ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; pharmacokinetics ; Glucosides ; administration & dosage ; blood ; pharmacokinetics ; Isoflavones ; administration & dosage ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Wistar

To establish a new method for simultaneously determining the content of five gingerol compounds in different processing degrees of ginger charcoal and PCA principal component analysis was conducted for analysis. Samples were analyzed on Ultimate TM XB-C18 column (4.6 mm x 250 mm, 5 μm) , with acetonitrile (A) -0.1% phosphoric acid solution (B) as mobile phase for gradient elution. Detection wavelength was set at 280 nm. The flow rate was 0.6 mL x min(-1) and the column temperature was 30 degrees C. The five compounds were separated well and showed good linearity (r ≥ 0.999 7) within the concentration ranges tested. The average value for recoveries was between 98.86% - 101.5% (RSD 1.4% - 2.9%). The contents of five compounds showed difference among different processing degrees of ginger charcoal. Zingiberone had the highest content in the standard carbon, and the content of gingerol was decreased as the deepening of processing degree. Different processing degrees of ginger charcoal were classified into three groups with PCA, and provided scientific basis for establishing the quality standards of ginger charcoal.
Catechols ; chemistry ; Charcoal ; chemistry ; Chemistry, Pharmaceutical ; methods ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Fatty Alcohols ; chemistry ; Ginger ; chemistry ; Principal Component Analysis ; methods

This study aims to establish an HPLC method for simultaneous determination of gastrodin and eight nucleosides and nucleobases components in Gastrodia elata. The separation was carried out on an Agilent Zorbax Bonus-RP (4.6 mm x 250 mm, 5 μm) column with a methanol-(0.04% acetic acid) water solution gradient elution program at a flow rate of 1.0 mL x min(-1). The column temperature was 36 degrees C, and the detection wavelength was 254 nm. The volume of injection was 20 μL. The nine components including gastrodin, cytosine, uracil, cytosine, adenine, thymine, uridine, guanosine and adenosine were well separated. The calibration curve was well linear in the range of 2.04-262.00 mg x L(-1), 0.20-24.67 mg x L(-1), 0.18-23.75 mg x L(-1), 0.20-25.83 mg x L(-1), 0.20-26.67 mg x L(-1), 0.16-20.00 mg x L(-1), 0.22-27.71 mg x L(-1), 0.20-24.29 mg x L(-1), 0.24-30.58 mg x L(-1), respectively, and the correlation coefficient was between 0.998 9-0.999 9. The average recovery of gastrodin and eight nucleosides and nucleobases were 96.4%-99.6%, RSD less than 2.7% (n = 6). The contents of gastrodin in all the seven Tibet cultured Gastrodia elata samples were over 2 mg x g(-1). Further, all samples contain higher contents of adenosine, guanosine, uridine and cytidine compared to low contents of cytosine, uracil, adenine and thymine. The established method is accurate, reproducible and suitable for the determination of gastrodin and eight nucleosides and nucleobases comppnents in Gastrodia elata.
Benzyl Alcohols ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Gastrodia ; chemistry ; Glucosides ; analysis ; Nucleosides ; analysis ; Nucleotides ; analysis

The hydrophilicity of the normal decoction pieces (NDP) of Indigo Naturalis is not good, therefore, it is not suit for decoctions. In this paper, powder modification technology is used and some NDP and alcohol are ground together in the vibromill to prepare the hydrophilic decoction pieces (HDP) of Indigo Naturalis. Initially, the properties of NDP, ultrafine decoction pieces (UDP) and HDP are compared, the hydrophilicity of UDP was promoted slightly, that of HDP is promoted dramatically. Then, three batches of Indigo Naturalis are prepared to HDP separately, but there is no obvious difference in the contact angle. Furthermore, the size distribution, surface area and micro-shape of HDP are bigger than that of UDP and smaller than NDP. The contents of indigo and indirubin in three decoction pieces are the same, as well as the species of inorganic substance, although there is a little difference in the proportion of five inorganic substances. The fact suggests the change of physical state and the qualitative and quantitative change of organism and inorganic substances are not the main factors to influence the hydrophilicity. In addition, hydroxyl, methylene and methyl can be identified at the wavenumber of 3 356 cm(-1) and 1 461 cm(-1) in infrared spectrum; the content of alcohol in HDP is 0.67% measured by gas chromatogram. The stability of HDP in the heating condition is studied, the fact suggests the hydrophilic effect of HDP at 40 degrees C is relatively stable. All above research suggests that the alcohol is the main factor to influence the hydrophilicity and maybe the intermolecular force which fixed alcohol molecule on the surface of Indigo Naturalis is the basic principle to produce the hydrophilicity.
Acanthaceae ; chemistry ; Alcohols ; analysis ; Hydrophobic and Hydrophilic Interactions ; Indigo Carmine ; analysis ; chemistry ; isolation & purification ; Indoles ; analysis ; Isatis ; chemistry ; Microscopy, Electron, Scanning ; Particle Size ; Plants, Medicinal ; chemistry ; Polygonum ; chemistry ; Powders ; Surface Properties ; Technology, Pharmaceutical ; methods ; X-Ray Diffraction

OBJECTIVETo develop an HPLC method for determining four components in Sini Tan, benzoylmesaconine, liquiritin, glycyrrhizic acid and 6-gingerol.

METHODThe Hypersil BDS column was adopted with gradient elution program at a flow rate of 1.0 mL x min(-1) and the detection wavelength of 235 nm.

RESULTBenzoylmesaconine, liquiritin, glycyrrhizic acid and 6-gingerol showed good separation, with the linear range of 0.006-0.12, 0.021-0.42, 0.012-0.24 and 0.018-0.36 g x L(-1), respectively. Their average recoveries were 99.3%, 96.9%, 100% and 100%, respectively; and RSD of the above four components were 1.5%, 0.6%, 1.3% and 2.1%, respectively.

CONCLUSIONThe method is proved to be so easy and accurate and practical that it can be used to determine the four components in Sini Tang.

Aconitine ; analogs & derivatives ; analysis ; isolation & purification ; Anti-Inflammatory Agents ; analysis ; isolation & purification ; Catechols ; analysis ; isolation & purification ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Fatty Alcohols ; analysis ; isolation & purification ; Flavanones ; analysis ; isolation & purification ; Glucosides ; analysis ; isolation & purification ; Glycyrrhizic Acid ; analysis ; isolation & purification ; Medicine, Chinese Traditional ; Reproducibility of Results

OBJECTIVETo discuss the synergistic mechanism of compatible use of two medicinal herbs, Zingiber offiicinale and Aconitum cainichaeli, by determining single decoction of Z. offiicinale and four gingerols (6-gingerol, 8-gingerol, 6-shogaol, 10-gingerol) contained in compound decoction of Z. offiicinale and A. cainichaeli of different compatibility ratio using HPLC.

METHODKromasil-C18 column (4.6 mm x 250 mm, 5 microm) was adopted. The mobile phase was acetonitrile (B) and 0.1% aqueous acetic acid (A) for gradient elution (0-30 min, 40%-90% B; 30-35 min, 90%-40% B). The flow rate was 1.0 mL x min(-1). The detection wavelength was set at 275 nm. The column temperature was 30 degrees C.

RESULTThe four gingerols were in baseline separation, with a good linearity (r > 0.999), an average recovery of 100.9% -103.5% and RSD < 3.0%. Compared with the single decoction of Z. offiicinale, the content of gingerols in the compound decoction of Z. offiicinale and A. cainichaeli was on the rise and in direct proportion with the increase in the volume of A. cainichaeli.

CONCLUSIONThe synergistic mechanism of the compatibility of Z. offiicinale and A. cainichaeli can be proved with the increased release of gingerols from Z. offiicinale.

Aconitum ; Catechols ; analysis ; Drug Compounding ; Drug Synergism ; Fatty Alcohols ; analysis ; Ginger ; chemistry

OBJECTIVETo establish an HPLC method for simultaneous determination of panaxynol and panaxydol from the fibrous root of Panax ginseng.

METHODThe analysis was performed on Elite C18 column (4.6 mm x 150 mm, 5 microm) with mobile phase gradient of CH3CN-water at a flow rate of 1.0 mL x min(-1). The detection wavelength was 230 nm, and the detection temperature was ambient.

RESULTThe linear range were 0.70-3.50 microg (r = 0.9995) for panaxynol, and 0.64-3.20 microg (r = 0.9999) for panaxydol. The average recoveries were 99.1% (RSD 1.7%) and 99.3% (RSD 1.2%), respectively.

CONCLUSIONThe HPLC method is simple, rapid and reproducible, which can be used for the quality control of the fibrous root of P. ginseng.

Chromatography, High Pressure Liquid ; methods ; Diynes ; analysis ; Drugs, Chinese Herbal ; analysis ; Fatty Alcohols ; analysis ; Panax ; chemistry ; Plant Roots ; chemistry