Sample records for 2-3-pentanedione

  1. Respiratory and olfactory cytotoxicity of inhaled 2,3-pentanedione in Sprague-Dawley rats.

    Hubbs, Ann F; Cumpston, Amy M; Goldsmith, W Travis; Battelli, Lori A; Kashon, Michael L; Jackson, Mark C; Frazer, David G; Fedan, Jeffrey S; Goravanahally, Madhusudan P; Castranova, Vincent; Kreiss, Kathleen; Willard, Patsy A; Friend, Sherri; Schwegler-Berry, Diane; Fluharty, Kara L; Sriram, Krishnan


    Flavorings-related lung disease is a potentially disabling disease of food industry workers associated with exposure to the α-diketone butter flavoring, diacetyl (2,3-butanedione). To investigate the hypothesis that another α-diketone flavoring, 2,3-pentanedione, would cause airway damage, rats that inhaled air, 2,3-pentanedione (112, 241, 318, or 354 ppm), or diacetyl (240 ppm) for 6 hours were sacrificed the following day. Rats inhaling 2,3-pentanedione developed necrotizing rhinitis, tracheitis, and bronchitis comparable to diacetyl-induced injury. To investigate delayed toxicity, additional rats inhaled 318 (range, 317.9-318.9) ppm 2,3-pentanedione for 6 hours and were sacrificed 0 to 2, 12 to 14, or 18 to 20 hours after exposure. Respiratory epithelial injury in the upper nose involved both apoptosis and necrosis, which progressed through 12 to 14 hours after exposure. Olfactory neuroepithelial injury included loss of olfactory neurons that showed reduced expression of the 2,3-pentanedione-metabolizing enzyme, dicarbonyl/L-xylulose reductase, relative to sustentacular cells. Caspase 3 activation occasionally involved olfactory nerve bundles that synapse in the olfactory bulb (OB). An additional group of rats inhaling 270 ppm 2,3-pentanedione for 6 hours 41 minutes showed increased expression of IL-6 and nitric oxide synthase-2 and decreased expression of vascular endothelial growth factor A in the OB, striatum, hippocampus, and cerebellum using real-time PCR. Claudin-1 expression increased in the OB and striatum. We conclude that 2,3-pentanedione is a respiratory hazard that can also alter gene expression in the brain. PMID:22894831

  2. Development of Air Sampling and Analytical Methods for Acetoin, Diacetyl, and 2,3-Pentanedione

    Takaku-Pugh, Sayaka


    Acetoin, diacetyl, and 2,3-pentanedione are artificial butter flavoring ingredients. Occupational exposures to diacetyl are associated with severe respiratory disease including bronchiolitis obliterans. Dynamic and passive air sampling and analytical methods were developed for simultaneous sampling of these ketones using 10 % (w/w) O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine hydrochloride (PFBHA) on Tenax TA (80/100 mesh). PFBHA O-oximes of the ketones were synthesized with above 95 % pur...

  3. Low-Pressure Photolysis of 2,3-Pentanedione in Air: Quantum Yields and Reaction Mechanism.

    Bouzidi, Hichem; Djehiche, Mokhtar; Gierczak, Tomasz; Morajkar, Pranay; Fittschen, Christa; Coddeville, Patrice; Tomas, Alexandre


    Dicarbonyls in the atmosphere mainly arise from secondary sources as reaction products in the degradation of a large number of volatile organic compounds (VOC). Because of their sensitivity to solar radiation, photodissociation of dicarbonyls can dominate the fate of these VOC and impact the atmospheric radical budget. The photolysis of 2,3-pentanedione (PTD) has been investigated for the first time as a function of pressure in a static reactor equipped with continuous wave cavity ring-down spectroscopy to measure the HO2 radical photostationary concentrations along with stable species. We showed that (i) Stern-Volmer plots are consistent with low OH-radical formation yields in RCO + O2 reactions, (ii) the decrease of the photodissociation rate due to pressure increase from 26 to 1000 mbar is of about 30%, (iii) similarly to other dicarbonyls, the Stern-Volmer analysis shows a curvature at the lower pressure investigated, which may be assigned to the existence of excited singlet and triplet PTD states, (iv) PTD photolysis at 66 mbar leads to CO2, CH2O and CO with yields of (1.16 ± 0.04), (0.33 ± 0.02) and (0.070 ± 0.005), respectively, with CH2O yield independent of pressure up to 132 mbar and CO yield in agreement with that obtained at atmospheric pressure by Bouzidi et al. (2014), and (v) the PTD photolysis mechanism remains unchanged between atmospheric pressure and 66 mbar. As a part of this work, the O2 broadening coefficient for the absorption line of HO2 radicals at 6638.21 cm(-1) has been determined (γO2 = 0.0289 cm(-1) atm(-1)). PMID:26608471

  4. Diacetyl and 2,3-pentanedione exposure of human cultured airway epithelial cells: Ion transport effects and metabolism of butter flavoring agents.

    Zaccone, Eric J; Goldsmith, W Travis; Shimko, Michael J; Wells, J R; Schwegler-Berry, Diane; Willard, Patsy A; Case, Shannon L; Thompson, Janet A; Fedan, Jeffrey S


    Inhalation of butter flavoring by workers in the microwave popcorn industry may result in “popcorn workers' lung.” In previous in vivo studies rats exposed for 6 h to vapor from the flavoring agents, diacetyl and 2,3-pentanedione, acquired flavoring concentration-dependent damage of the upper airway epithelium and airway hyporeactivity to inhaled methacholine. Because ion transport is essential for lung fluid balance,we hypothesized that alterations in ion transport may be an early manifestation of butter flavoring-induced toxicity.We developed a system to expose cultured human bronchial/tracheal epithelial cells (NHBEs) to flavoring vapors. NHBEs were exposed for 6 h to diacetyl or 2,3-pentanedione vapors (25 or ≥ 60 ppm) and the effects on short circuit current and transepithelial resistance (Rt) were measured. Immediately after exposure to 25 ppm both flavorings reduced Na+ transport,without affecting Cl- transport or Na+,K+-pump activity. Rt was unaffected. Na+ transport recovered 18 h after exposure. Concentrations (100-360 ppm) of diacetyl and 2,3-pentanedione reported earlier to give rise in vivo to epithelial damage, and 60 ppm, caused death of NHBEs 0 h post-exposure. Analysis of the basolateral medium indicated that NHBEs metabolize diacetyl and 2,3-pentanedione to acetoin and 2-hydroxy-3-pentanone, respectively. The results indicate that ion transport is inhibited transiently in airway epithelial cells by lower concentrations of the flavorings than those that result in morphological changes of the cells in vivo or in vitro. PMID:26454031

  5. 76 FR 1434 - Request for Information on 2,3-Pentanedione and Other Alpha-Diketones Used As Diacetyl Substitutes


    ... Occupational Safety and Health (NIOSH) of the Centers for Disease Control and Prevention (CDC) intends to... Safety and Health (NIOSH) of the Centers for Disease Control and Prevention (CDC), Department of Health... propionyl) and other alpha-diketones and develop appropriate communication documents, such as a...

  6. Influence of valine and other amino acids on total diacetyl and 2,3-pentanedione levels during fermentation of brewer’s wort

    Krogerus, Kristoffer; Gibson, Brian R.


    Undesirable butter-tasting vicinal diketones are produced as by-products of valine and isoleucine biosynthesis during wort fermentation. One promising method of decreasing diacetyl production is through control of wort valine content since valine is involved in feedback inhibition of enzymes controlling the formation of diacetyl precursors. Here, the influence of valine supplementation, wort amino acid profile and free amino nitrogen content on diacetyl formation during wort fermentation with...

  7. 77 FR 21777 - Expanded Charge for Peer Review of the NIOSH document Titled: “Criteria for a Recommended...


    ...The National Institute for Occupational Safety and Health (NIOSH) of the Centers for Disease Control and Prevention (CDC) is undergoing peer review for the draft document, ``Criteria for a Recommended Standard: Occupational Exposure to Diacetyl and 2,3- pentanedione.'' NIOSH held a public meeting on August 26, 2011 in Washington, DC [76 FR 44338] to discuss and obtain comments on the draft......

  8. Inhibition of LuxS by S-Ribosylhomocysteine Analogues Containing a [4-Aza]Ribose Ring

    Malladi, Venkata L.A.; Sobczak, Adam J.; Meyer, Tiffany M.; Pei, Dehua; Wnuk, Stanislaw F.


    LuxS (S-ribosylhomocysteinase) catalyzes the cleavage of the thioether linkage of S- ribosylhomocysteine (SRH) to produce homocysteine and 4,5-dihydroxy-2,3-pentanedione (DPD), the precursor to a small signaling molecule that mediates interspecies bacterial communication called autoinducer 2 (AI-2). Inhibitors of LuxS should interfere with bacterial interspecies communication and potentially provide a novel class of antibacterial agents. In this work, SRH analogues containing substitution of ...

  9. Glycation Reactivity of a Quorum-Sensing Signaling Molecule.

    Tsuchikama, Kyoji; Gooyit, Major; Harris, Tyler L; Zhu, Jie; Globisch, Daniel; Kaufmann, Gunnar F; Janda, Kim D


    Reported herein is that (4S)-4,5-dihydroxy-2,3-pentanedione (DPD) can undergo a previously undocumented non-enzymatic glycation reaction. Incubation of DPD with viral DNA or the antibiotic gramicidin S resulted in significant biochemical alterations. A protein-labeling method was consequently developed that facilitated the identification of unrecognized glycation targets of DPD in a prokaryotic system. These results open new avenues toward tracking and understanding the fate and function of the elusive quorum-sensing signaling molecule. PMID:26890076

  10. α,β-Dicarbonyl reduction is mediated by the Saccharomyces Old Yellow Enzyme.

    van Bergen, Barry; Cyr, Normand; Strasser, Rona; Blanchette, Maxime; Sheppard, John D; Jardim, Armando


    The undesirable flavor compounds diacetyl and 2,3-pentanedione are vicinal diketones (VDKs) formed by extracellular oxidative decarboxylation of intermediate metabolites of the isoleucine, leucine and valine (ILV) biosynthetic pathway. These VDKs are taken up by Saccharomyces and enzymatically converted to acetoin and 3-hydroxy-2-pentanone, respectively. Purification of a highly enriched diacetyl reductase fraction from Saccharomyces cerevisiae in conjunction with mass spectrometry identified Old Yellow Enzyme (Oye) as an enzyme capable of catalyzing VDK reduction. Kinetic analysis of recombinant Oye1p, Oye2p and Oye3p isoforms confirmed that all three isoforms reduced diacetyl and 2,3-pentanedione in an NADPH-dependent reaction. Transcriptomic analysis of S. cerevisiae (ale) and S. pastorianus (lager) yeast during industrial fermentations showed that the transcripts for OYE1, OYE2, arabinose dehydrogenase (ARA1), α-acetolactate synthase (ILV2) and α-acetohydroxyacid reductoisomerase (ILV5) were differentially regulated in a manner that correlated with changes in extracellular levels of VDKs. These studies provide insights into the mechanism for reducing VDKs and decreasing maturation times of beer which are of commercial importance. PMID:27400981

  11. The response of Serratia marcescens JG to environmental changes by quorum sensing system.

    Sun, Shu-Jing; Liu, Hui-Jun; Weng, Cai-Hong; Lai, Chun-Fen; Ai, Liu-Ying; Liu, Yu-Chen; Zhu, Hu


    Many bacterial cells are known to regulate their cooperative behaviors and physiological processes through a molecular mechanism called quorum sensing. Quorum sensing in Serratia marcescens JG is mediated by the synthesis of autoinducer 2 (AI-2) which is a furanosyl borate diester. In this study, the response of quorum sensing in S. marcescens JG to environment changes such as the initial pH, carbon sources and boracic acid was investigated by a bioreporter and real-time PCR analysis. The results show that glucose can affect AI-2 synthesis to the greatest extent, and 2.0 % glucose can stimulate S. marcescens JG to produce more AI-2, with a 3.5-fold increase in activity compared with control culture. Furthermore, the response of quorum sensing to changes in glucose concentration was performed by changing the amount of luxS RNA transcripts. A maximum of luxS transcription appeared during the exponential growth phase when the glucose concentration was 20.0 g/L. AI-2 production was also slightly impacted by the low initial pH. It is significant for us that the addition of boracic acid at microdosage (0.1-0.2 g/L) can also induce AI-2 synthesis, which probably demonstrated the feasible fact that the 4,5-dihydroxy-2, 3-pentanedione cyclizes by the addition of borate and the loss of water, is hydrated and is converted to the final AI-2 in S. marcescens JG. PMID:27020680

  12. Synthesis, characterization, and tyrosinase biomimetic catalytic activity of copper(II) complexes with schiff base ligands derived from α-diketones with 2-methyl-3-amino-(3 H)-quinazolin-4-one

    Ramadan, Abd El-Motaleb M.; Ibrahim, Mohamed M.; Shaban, Shaban Y.


    A template condensation of α-diketones (biacetyl, benzile and 2,3-pentanedione) with 2-methyl-3-amino-(3 H)-quinazolin-4-one (AMQ) in the presence of CuX 2 (X = Cl -, Br -, NO3- or ClO4-) resulted in the formation of tetradentate Schiff base copper(II) complexes of the type [CuLX]X and [CuL]X 2. Structural characterization of the complex species was achieved by several physicochemical methods, namely elemental analysis, electronic spectra, IR, ESR, molar conductivity, thermal analysis (TAG & DTG), and magnetic moment measurements. The stereochemistry, the nature of the metal chelates, and the catalytic reactivity are markedly dependent upon the type of counter anions and the ligand substituent within the carbonyl moiety. A square planar monomeric structure is proposed for the perchlorate, nitrate, and bromide complexes, in which the counter anions are loosely bonded to copper(II) ion. For the chloride complexes, the molar conductivities and the spectral data indicated that they have square-pyramidal environments around copper(II) center. The reported copper(II) complexes exhibit promising tyrosinase catalytic activity towards the hydroxylation of phenol followed by the aerobic oxidation of the resulting catechol. A linear correlation almost exists between the catalytic reactivity and the Lewis-acidity of the central copper(II) ion created by the donating properties of the parent ligand. The steric considerations could be accounted to clarify the difference in the catalytic activity of these functional models.

  13. IDENTIFIKASI CHARACTER IMPACT ODORANTS BUAH KAWISTA (Feronia Limonia [Identification of Character Impact Odorants of Wood Apple Fruit (Feronia Limonia

    Anton Apriyantono1


    Full Text Available The volatiles of the kawista fruit (wood apple were analyzed by gas chromatography (GC and a combined gas chromatography-mass spectrometer (GC-MS. Character impact odorants of the fruits were systematically characterized by aroma extract dilution analysis (AEDA with GC-Olfactometry (GC-O. A total of 75 compounds were identified, including 28 esters, 11 alcohols, 10 aldehydes, 1 acetal, 10 ketones, 4 lactones, 1 heterocyclic, 4 aliphatic hydrocarbons, 1 furan and 5 acids. However, only 44 volatiles were detected by GC-O. Among these, compounds with the most impact were ethyl butyrate (fruity, sweet, banan-likeand methyl butyrate (fruit, sour with a flavor dilution factor of 256 and 64, respectively. Based on AEDA results, butyric acid, 3-mathyl valeric acid, 1-octen-3-ol, pentyl isobutyrate, 2-ethyl hexanoic acid, ethyl octanoate, gamma-decalactone, 2,3-pentanedione, 3-octanone, 5-methyl-3-heptanone, 9-methyl-5-undecene and (E-2-hexenyl butyrate seem to contribute to kawista fruit flavor

  14. Sampling and Analytical Method for Alpha-Dicarbonyl Flavoring Compounds via Derivatization with o-Phenylenediamine and Analysis Using GC-NPD

    Pendergrass, Stephanie M.; Cooper, Jeffrey A.


    A novel methodology is described for the sampling and analysis of diacetyl, 2,3-pentanedione, 2,3-hexanedione, and 2,3-heptanedione. These analytes were collected on o-phenylenediamine-treated silica gel tubes and quantitatively recovered as the corresponding quinoxaline derivatives. After derivatization, the sorbent was desorbed in 3 mL of ethanol solvent and analyzed using gas chromatography/nitrogen-phosphorous detection (GC/NPD). The limits of detection (LOD) achieved for each analyte were determined to be in the range of 5–10 nanograms/sample. Evaluation of the on-tube derivatization procedure indicated that it is unaffected by humidities ranging from 20% to 80% and that the derivatization procedure was quantitative for analyte concentrations ranging from 0.1 μg to approximately 500 μg per sample. Storage stability studies indicated that the derivatives were stable for 30 days when stored at both ambient and refrigerated temperatures. Additional studies showed that the quinoxaline derivatives were quantitatively recovered when sampling up to a total volume of 72 L at a sampling rate of 50 cc/min. This method will be important to evaluate and monitor worker exposures in the food and flavoring industry. Samples can be collected over an 8-hour shift with up to 288 L total volume collected regardless of time, sampling rate, and/or the effects of humidity. PMID:27247828

  15. Exploratory and discriminative studies of commercial processed Brazilian coffees with different degrees of roasting and decaffeinated

    Juliano Souza Ribeiro


    Full Text Available The fingerprints of the volatile compounds of 21 commercial Brazilian coffee samples submitted to different industrial processing i.e. decaffeinated or different roasting degrees (traditional and dark were studied. The volatiles were collected by headspace solid phase microextraction (HS-SPME and analyzed by GC-FID and GC-MS. The chromatographic data matrices (fingerprints obtained were explored by the principal component analysis (PCA and partial least squares - discriminative analysis (PLS-DA. Initially the chromatographic profiles were aligned by the algorithm correlation optimized warping (COW. The PCA showed the discrimination of the decaffeinated coffees from the others with both the SPME fibres used. This separation probably occurred due to the loss of some volatile precursors during the decaffeination process, such as sucrose. For both the fibres tested, PDMS/DVB and CX / PDMS SPME, the PLS-DA models correctly classified 100% of the samples according to their roasting degree: (medium and dark, the main differences being the concentrations of some of the volatile compounds such as 2-methyl furan, 2-methylbutanal, 2,3-pentanedione, pyrazine, 2-carboxyaldehyde pyrrole, furfural and 2-furanmethanol.

  16. Volatile compounds of some popular Mediterranean seafood species



    Full Text Available The volatile compounds of highly commercialised fresh Mediterranean seafood species, including seven fish (sand-smelt Atherina boyeri, picarel Spicara smaris, hake Merluccius merluccius, pilchard Sardina pilchardus, bogue Boobps boops, anchovy Engraulis encrasicolus and striped-mullet Mullus barbatus, squid (Loligo vulgaris, shrimp (Parapenaeus longirostris and mussel (Mytilus galloprovincialis, were evaluated by simultaneous steam distillation-extraction and subsequent GC-MS analysis. A total of 298 volatile compounds were detected. The mussels contained the highest total concentration of volatile compounds, while pilchard among fish species contained the highest number and concentrations of volatile compounds. Individual patterns of volatile compounds have been distinguished. The fish species when compared to the shellfish species studied, contained 6 to 30 times more 1-penten-3-ol, higher quantities of 2-ethylfuran, and 2,3-pentanedione, which was absent from the shellfish species. Pilchard is characterized by a high concentration of alcohols, shrimps by the high presence of amines and S-compounds, while mussels by high amounts of aldehydes, furans, and N-containing compounds (pyridine, pyrazines and pyrrols. The fatty acid-originating carbonyl compounds in fish seem to be related to the species’ fat content.

  17. Odor-Active (E -4-Methyl-3-hexenoic Acid in Roasted Coffee Generated in the Maillard Reactions of L-Isoleucine with Sugars

    Hironari Miyazato


    Full Text Available This study describes a novel odor-active unsaturated aliphatic acid in roasted Brazilian Arabica coffee. (E-4-Methyl-3-hexenoic acid -4M3H, which is responsible for a sweaty odor, was identified for the first time using Gas Chromatography-Mass Spectrometry/Olfactometry (GC-MS/O, Aroma Extract Dilution Analysis (AEDA and Multidimensional Gas Chromatography-Mass Spectrometry (MDGC-MS; its (Z -isomer was also determined. Additionally, we conducted a model formation experiment to detect the presence of 4M3H in coffee and found that it may be produced nonstereoselectively in the Maillard reactions of L-isoleucine with sugars (xylose, fructose, glucose, rhamnose and sucrose. We also found that 2-methylbutanal derived from L-isoleucine and sugar degradation compounds such as α-dicarbonyl compounds (glyoxal, 2-oxopropanal, 2, 3-butanedione and 2, 3-pentanedione and α-hydroxy ketones (1-hydroxy-2-propanone and 3-hydroxy-2-butanone are key factors in the formation of 4M3H. Based on these results, we propose a series of potential nonstereoselective formation pathways for 4M3H.

  18. Proton transfer reaction time-of-flight mass spectrometry monitoring of the evolution of volatile compounds during lactic acid fermentation of milk.

    Soukoulis, Christos; Aprea, Eugenio; Biasioli, Franco; Cappellin, Luca; Schuhfried, Erna; Märk, Tilmann D; Gasperi, Flavia


    We apply, for first time, the recently developed proton transfer reaction time-of-flight mass spectrometry (PTR-TOF-MS) apparatus as a rapid method for the monitoring of lactic acid fermentation (LAF) of milk. PTR-TOF-MS has been proposed as a very fast, highly sensitive and versatile technique but there have been no reports of its application to dynamic biochemical processes with relevance to the food industry. LAF is a biochemical-physicochemical dynamic process particularly relevant for the dairy industry as it is an important step in the production of many dairy products. Further, LAF is important in the utilization of the by-products of the cheese industry, such as whey wastewaters. We show that PTR-TOF-MS is a powerful method for the monitoring of major volatile organic chemicals (VOCs) formed or depleted during LAF, including acetaldehyde, diacetyl, acetoin and 2-propanone, and it also provides information about the evolution of minor VOCs such as acetic acid, 2,3-pentanedione, ethanol, and off-flavor related VOCs such as dimethyl sulfide and furfural. This can be very important considering that the conventional measurement of pH decrease during LAF is often ineffective due to the reduced response of pH electrodes resulting from the formation of protein sediments. Solid-phase microextraction gas chromatography/mass spectrometry (SPME-GC/MS) data on the inoculated milk base and final fermented product are also presented to supporting peak identification. We demonstrate that PTR-TOF-MS can be used as a rapid, efficient and non-invasive method for the monitoring of LAF from headspace, supplying important data about the quality of the final product and that it may be used to monitor the efficacy of manufacturing practices. PMID:20552689

  19. Processing the Interspecies Quorum-sensing Signal Autoinducer-2 (AI-2)

    J Marques; P Lamosa; C Russell; R Ventura; C Maycock; M Semmelhack; S Miller; K Xavier


    The molecule (S)-4,5-dihydroxy-2,3-pentanedione (DPD) is produced by many different species of bacteria and is the precursor of the signal molecule autoinducer-2 (AI-2). AI-2 mediates interspecies communication and facilitates regulation of bacterial behaviors such as biofilm formation and virulence. A variety of bacterial species have the ability to sequester and process the AI-2 present in their environment, thereby interfering with the cell-cell communication of other bacteria. This process involves the AI-2-regulated lsr operon, comprised of the Lsr transport system that facilitates uptake of the signal, a kinase that phosphorylates the signal to phospho-DPD (P-DPD), and enzymes (like LsrG) that are responsible for processing the phosphorylated signal. Because P-DPD is the intracellular inducer of the lsr operon, enzymes involved in P-DPD processing impact the levels of Lsr expression. Here we show that LsrG catalyzes isomerization of P-DPD into 3,4,4-trihydroxy-2-pentanone-5-phosphate. We present the crystal structure of LsrG, identify potential catalytic residues, and determine which of these residues affects P-DPD processing in vivo and in vitro. We also show that an lsrG deletion mutant accumulates at least 10 times more P-DPD than wild type cells. Consistent with this result, we find that the lsrG mutant has increased expression of the lsr operon and an altered profile of AI-2 accumulation and removal. Understanding of the biochemical mechanisms employed by bacteria to quench signaling of other species can be of great utility in the development of therapies to control bacterial behavior.

  20. Enantioselective Synthesis of Vicinal (R,R)-Diols by Saccharomyces cerevisiae Butanediol Dehydrogenase.

    Calam, Eduard; González-Roca, Eva; Fernández, M Rosario; Dequin, Sylvie; Parés, Xavier; Virgili, Albert; Biosca, Josep A


    Butanediol dehydrogenase (Bdh1p) from Saccharomyces cerevisiae belongs to the superfamily of the medium-chain dehydrogenases and reductases and converts reversibly R-acetoin and S-acetoin to (2R,3R)-2,3-butanediol and meso-2,3-butanediol, respectively. It is specific for NAD(H) as a coenzyme, and it is the main enzyme involved in the last metabolic step leading to (2R,3R)-2,3-butanediol in yeast. In this study, we have used the activity of Bdh1p in different forms-purified enzyme, yeast extracts, permeabilized yeast cells, and as a fusion protein (with yeast formate dehydrogenase, Fdh1p)-to transform several vicinal diketones to the corresponding diols. We have also developed a new variant of the delitto perfetto methodology to place BDH1 under the control of the GAL1 promoter, resulting in a yeast strain that overexpresses butanediol dehydrogenase and formate dehydrogenase activities in the presence of galactose and regenerates NADH in the presence of formate. While the use of purified Bdh1p allows the synthesis of enantiopure (2R,3R)-2,3-butanediol, (2R,3R)-2,3-pentanediol, (2R,3R)-2,3-hexanediol, and (3R,4R)-3,4-hexanediol, the use of the engineered strain (as an extract or as permeabilized cells) yields mixtures of the diols. The production of pure diol stereoisomers has also been achieved by means of a chimeric fusion protein combining Fdh1p and Bdh1p. Finally, we have determined the selectivity of Bdh1p toward the oxidation/reduction of the hydroxyl/ketone groups from (2R,3R)-2,3-pentanediol/2,3-pentanedione and (2R,3R)-2,3-hexanediol/2,3-hexanedione. In conclusion, Bdh1p is an enzyme with biotechnological interest that can be used to synthesize chiral building blocks. A scheme of the favored pathway with the corresponding intermediates is proposed for the Bdh1p reaction. PMID:26729717

  1. 东北特产许氏大酱中挥发性成分的提取与分析%Extraction and Analysis of Volatile Flavor Constituents of XuShi Soybean Paste as Northeast Special Product

    孙洁雯; 杨克玉; 李燕敏; 刘玉平


    The characteristic aroma components in XuShi soybean paste as Northeast Special Product were extracted by simultaneous distillation-extraction (SDE). The operating conditions of SDE were optimized for their effect on the analysis of flavors , including amount of raw materials and extraction time. Based on the calculated retention indices and RI qualitative analysis , 33 compounds were identified in XuShi soybean paste , Their total content in XuShi soybean paste was 24.41μg/g. These compounds included 13 esters(8.24μg/g), 5 aldehydes (11.74μg/g), 4 heterocyclic compounds (0.87μg/g), 3 ketones (0.85μg/g), 2 sulfur-containing ethers(0.36μg/g),6 other compounds(2.35μg/g). The major aroma components were 3-Methyl-butanal, 2-Methyl-butanal, ethyl acetate. Hexanal, benzeneacetaldehyde, 3-Octanone , dimethyl disulfide, 2,3-Pentanedione , 2-Pentyl furan made a greater contribution to the odor characteristics of XuShi soybean paste.%为了研究作为东北特产的许氏大酱的挥发性成分, 采用同时蒸馏萃取法对许氏大酱挥发性成分进行提取,并考察了原料量和提取时间对结果的影响.提取物经气-质联用仪分析,结合计算保留指数和NIST11谱库检索,确定了许氏大酱中挥发性成分的种类,采用内标法确定了挥发性成分在许氏大酱中的含量.共鉴定出33种挥发性成分,在许氏大酱中含量为24.41μg/g,包括酯类13种(8.24μg/g)、醛类5种(11.74μg/g)、杂环类4种(0.87μg/g)、酮类3种(0.85μg/g)、含硫化合物2种(0.36μg/g)、其他类6种(2.35μg/g).其中3-甲基丁醛、2-甲基丁醛、乙酸乙酯含量较大,且香气活性值(OAV值)>1,且数值较高,属于许氏大酱特征性香气成分.此外,己醛、苯乙醛、3-辛酮、二甲基二硫、2,3-戊二酮、2-戊基呋喃虽含量较少,但OAV值>1,且数值较高,对许氏大酱整体香气贡献较大.

  2. 线虫转型发育和寄主识别的化学通讯研究进展%Research progress on chemical communication of development and host-finding of nematodes

    张宾; 胡春祥; 石进; 蒋丽雅; 汤宗斌; 石敬夫; 赵莉蔺


    , nucleotides, biotin and some volatiles (pyrazine, diacetyl, benzaldehyde, 2, 4, 5-trimethylthiazole, isoamylalcohol, 2, 3-pentanedione ) . Another example is Bursaphelenchus xylophilus, the pinewood nematode, which is a plant parasitic nematode that is transmitted via Monochamus beetles. The chemical relationships within this system has been under investigation for several years, but no conclusive results regarding the discrete signaling molecules involved and their roles within the system have been obtained to date. Electrophysiological analyses indicated that different responses to chemical signals are due to differences in the chemoreceptors of various species of nematodes, which are composed by amphid and phasmid. Specific chemoreceptors in the neurons recognize particular compounds, and the neurons work collectively to receive chemical signals, and then activate a response through signal transduction. Nematodes, especially plant parasitic nematodes, cause huge economic damages every year, as advances in the study of the chemosenation mechanism are made, more effective strategies based on chemical ecology may be devised, thus replacing traditional control methods utilized today.%线虫是一类低等无脊椎动物,在自然界分布很广.因为线虫通常生活在土壤或寄生物中,没有适宜的视觉或听觉系统,接收环境信号的重要途径就是借助于其精细的化学感受系统.研究表明,线虫能够通过识别挥发性物质来引导一系列行为:取食、交配、产卵和驱避有毒物质、避免高种群密度等.目前,对线虫化学感受机制的研究越来越被人们所重视,也取得了一些突破性进展.综合近年来已有的研究成果,从发育调控机制、寄主识别机制、化学感受机理等方面进行了详细系统的总结,并对未来研究和线虫防治进行了展望.