Acetate Molecules Research Articles

ReaxFF Molecular Dynamics Study on the Influence of Temperature on Adsorption, Desorption, and Decomposition at the Acetic Acid/Water/ZnO(101̅0) Interface Enabling Cold Sintering.

The reaction dynamics of a liquid-solid interface with the example of an acetic acid/water solution interacting with a ZnO(101̅0) surface was investigated using ReaxFF reactive force field-based molecular dynamics. The interactions were studied over a broad temperature range to assess the kinetics and reaction pathways. Two different acetic acid dissociation mechanisms are observed in the simulations: (1) deprotonation to surface cation, which produces a terminal hydroxyl and (2) deprotonation to a bridging hydroxyl, which results in water production. An increase in temperature promotes the dissociation of acetic acids and its adsorption to surface at first, but as the temperature increase continues, the surface coverage by acetates decreases due to evaporation from the surface or decomposition. The acetate decomposition starts with a nucleophilic attack of oxygen to methyl carbon and results in the production of carbon dioxide, which is consistent with experimental findings in the literature. The coverage of the surface by water molecules decreases as the system is heated up, which is also observed in other molecular dynamics studies. At elevated temperatures, acetate molecules are more stable than water molecules or bridging hydroxyls on the surface. These simulations validate the ReaxFF method for the water/organic mixture and metal oxide surface interactions and provide insights into structure and reactivity of aqueous solvents on metal oxide surfaces at elevated temperatures. Adsorption trends that are observed in this study are consistent with phenomenological Langmuir models. The reaction of acetic acid decomposition to smaller molecules such as CO2 and CH2O agrees with experimental observations. Understanding the details of these dynamic surface reactions are critical to understand important new cold sintering processes that utilize transient liquid and solid reactions, and the latter could be used to predict solvent selection for cold sintering.

Molecular dynamics investigation on the morphology of HNIW affected by the growth condition

ABSTRACTThe crystal morphology of Hexanitrohexaazaisowurtzitane (HNIW) in vacuum and ethyl acetate was investigated by attachment energy (AE) model through molecular dynamics (MD). The simulated solvent-affected outcome agrees well with the experimental crystal habit cultivated from ethyl acetate. The influence of model size on the modified AE was discussed, and a reasonable model size was obtained. The model size can be broadened to calculate the morphology of other compounds. The radial distribution function (RDF) analyses and diffusion coefficient analyses were performed to explore the adsorption and diffusion behaviors of ethyl acetate molecules on HNIW surfaces. The simulation results can provide some guidance for the further crystallization process of HNIW.

Trypanosoma brucei aquaglyceroporins mediate the transport of metabolic end-products: Methylglyoxal, D-lactate, L-lactate and acetate

Bloodstream forms of Trypanosoma (T.) brucei, the causative agent of African sleeping sickness, possess a highly active glycolysis, which generates as main end-products: pyruvate under aerobic conditions, and pyruvate and glycerol under anaerobic conditions. To secrete them into the extracellular milieu, the parasites have at least two main specific membrane proteins, the pyruvate transporter and the aquaglyceroporins However, there are several other minor products from the glycolysis that must be excreted by the parasites and whose exit pathway until now remained elusive. As aquaglyceroporins from T. brucei (TbAQP1, 2, and 3) show a wide permeability profile for small solutes, we decided to evaluate if these proteins allow the passage of methylglyoxal, L-lactate, D-lactate and acetate molecules. We expressed heterologously TbAQP1, 2, and 3 in aquaglyceroporin-null yeast cells or in Xenopus laevis oocytes and demonstrated that these channels are permeable for methylglyoxal, L-lactate, D-lactate and acetate. We further demonstrate that methylglyoxal is highly toxic for bloodstream forms of T. brucei, while L-lactate and D-lactate appear almost harmless. Additionally, we discuss all our findings in the light of the novel metabolic discoveries, putting in context the participation of TbAQP1, 2, 3, and other proteins in the excretion of unwanted metabolic end-products.

Theoretical Investigation of Spectroscopic, Electronic and Thermodynamic Properties of [2-Methoxy-4-(3-methyl-4,5-dihydro-1H-1,2,4-triazol-5-one-4-yl)-azomethinphenyl] Acetate Molecule

Theoretical Investigation of Spectroscopic, Electronic and Thermodynamic Properties of [2-Methoxy-4-(3-methyl-4,5-dihydro-1H-1,2,4-triazol-5-one-4-yl)-azomethinphenyl] Acetate Molecule

Functional definition of the two effector binding sites, the oligomerization and DNA binding domains of the Bacillus subtilis LysR-type transcriptional regulator AlsR.

The LysR-type transcriptional regulator (LTTR) AlsR from Bacillus subtilis activates the transcription of the alsSD operon encoding enzymes for acetoin formation in response to the presence of acetate. The structural basis for effector binding, oligomerization, DNA binding, higher ordered complex formation, DNA bending and transcriptional control by B. subtilis AlsR was functionally characterized. The binding of two molecules of acetate per molecule AlsR was determined. Acetate-dependent transcription complex formation was observed. A structural model of AlsR was used to identify the amino acid residues V98, S100, H147 of the binding site 1, which were experimentally verified. The second binding site formed by T193, V194, A196, T201 and L202 mediated high acetate responsive induction. Residues L124, E225 Q74, I79 and R111 contributed to dimerization of AlsR. A22, Q29, P30, S33, K37, L39, E46, R50 and R53 of the winged helix-turn-helix motif were important for promoter recognition. The DNA binding domain alone dimerized and effectively bound the promoter. The LTTR promoter elements RBS and ABS had to be localized on the same site of the DNA. Higher ordered complex formation resulted in bending of promoter DNA and transcriptional activation.

Cellulose acetate - essential oil nanocapsules with antimicrobial activity for biomedical applications.

This study aimed to obtain bioactive nanosystems by combining cellulose acetate with three selected essential oils (EOs) to create spherical nanocapsules (NCs) using the solvent/anti-solvent technique. The biological activity of the obtained NCs was promoted by the use of some antimicrobial EOs: Peppermint, Cinnamon and lemongrass which were grafted on the cellulose acetate molecules. Due to their chemistry, such as long hydrocarbon tails and heads with functional groups these EOs were playing also the role of surfactant-like substance facilitating the formation of NCs. A dispersion of NCs was obtained in water and various spectroscopy techniques used to examine their size, morphology and chemistry. Dynamic light scattering calculate the size of the NCs whereas scanning electron microscopy showed their morphology. Fluorescent microscopy and Raman spectroscopy proved the attachment of the EOs in the cellulose acetate molecules. The antimicrobial activity of the obtained nanomaterials was tested against four microbial strains (bacteria: Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and a yeast strain of Candida albicans). The obtained results demonstrated that such NCs can be used in a variety of applications including medical, pharmaceutical recipients and in household products for treating or preventing microbial colonization and biofilm development.

Growth of Dehalococcoides mccartyi species in an autotrophic consortium producing limited acetate.

The dechlorinating Dehalococcoides mccartyi species requires acetate as carbon source, but little is known on its growth under acetate limiting conditions. In this study, we observed growth and dechlorination of a D. mccartyi-containing mixed consortium in a fixed-carbon-free medium with trichloroethene in the aqueous phase and H2/CO2 in the headspace. Around 4mM formate was produced by day 40, while acetate was constantly below 0.05mM. Microbial community analysis of the consortium revealed dominance by D. mccartyi and Desulfovibrio sp. (57 and 22% 16S rRNA gene copies, respectively). From this consortium, Desulfovibrio sp. strain F1 was isolated and found to produce formate and acetate (1.2mM and 48µM, respectively, by day 24) when cultivated alone in the above mentioned medium without trichloroethene. An established co-culture of strain F1 and D. mccartyi strain 195 demonstrated that strain 195 could grow and dechlorinate using acetate produced by strain F1; and that acetate was constantly below 25µM in the co-culture. To verify that such low level of acetate is utilizable by D. mccartyi, we cultivated strain 195 alone under acetate-limiting conditions and found that strain 195 consumed acetate to below detection (5µM). Based on the acetate consumption and cell yield of D. mccartyi, we estimated that on average 1.2 × 108 acetate molecules are needed to supply carbon for one D. mccartyi cell. Our study suggests that Desulfovibrio may supply a steady but low amount of fixed carbon to dechlorinating bacteria, exhibiting important implications for natural bio-attenuation when fixed carbon is limited.

Mixed lanthanide coordination polymers for temperature sensing and enhanced NdIII NIR luminescence

Three LnIII coordination polymers, [Ln2L3(NO3)3]n·2CH3COOCH2CH3 (Ln = Nd (Nd2L3), Eu (Eu2L3), Tb (Tb2L3)), following coordination-assembly reactions involving a new bi-monodentate salicylamide ligand, L = 1,4-bis[(2΄-thenylaminoformyl)phenoxyl]methyl}-benzene, have been successfully synthesized. Among them, Nd2L3 and Tb2L3 were structurally authenticated by single-crystal X-ray diffraction and the results indicate both Nd2L3 and Tb2L3 are one-dimensional corrugated blockchains with LnIII containing 64-membered block loops filled by ethyl acetate molecules alternating in nearly orthogonal directions. For they are isomorphous, two binary co-doped lanthanide coordination polymers, Tb0.965Eu1.035L3 and Tb0.717Nd1.283L3 are obtained. Luminescent studies indicate that Tb0.965Eu1.035L3 exhibits a temperature recognition property with the significantly high sensitivity of 2.61% per K in the 296–386 K temperature range upon excitation at 345 nm as well as white emission when excited in the range of 370–390 nm. Tb0.717Nd1.283L3 shows enhanced NdIII-centred NIR luminescence and diminished TbIII-centred luminescence as compared to those of Nd2L3 and Tb2L3. The results presented herein show that the energy transfer from TbIII to both EuIII and NdIII has been successfully achieved in Tb0.965Eu1.035L3 and Tb0.717Nd1.283L3. Generally speaking, this work explores the potential applications of lanthanide-based coordination polymers based on multi-monodentate salicylamide ligand in the area of thermal sensing.

Spectroscopic evaluation of synthesized 5β-dihydrocortisol and 5β-dihydrocortisol acetate binding mechanism with human serum albumin and their role in anticancer activity

Our study focus on the biological importance of synthesized 5β-dihydrocortisol (Dhc) and 5β-dihydrocortisol acetate (DhcA) molecules, the cytotoxic study was performed on breast cancer cell line (MCF-7) normal human embryonic kidney cell line (HEK293), the IC50 values for MCF-7 cells were 28 and 25 μM, respectively, whereas no toxicity in terms of cell viability was observed with HEK293 cell line. Further experiment proved that Dhc and DhcA induced 35.6 and 37.7% early apoptotic cells and 2.5, 2.9% late apoptotic cells, respectively, morphological observation of cell death through TUNEL assay revealed that Dhc and DhcA induced apoptosis in MCF-7 cells. The complexes of HSA–Dhc and HSA–DhcA were observed as static quenching, and the binding constants (K) was 4.7 ± .03 × 104 M−1 and 3.9 ± .05 × 104 M−1, and their binding free energies were found to be −6.4 and −6.16 kcal/mol, respectively. The displacement studies confirmed that lidocaine 1.4 ± .05 × 104 M−1 replaced Dhc, and phenylbutazone 1.5 ± .05 × 104 M−1 replaced by DhcA, which explains domain I and domain II are the binding sites for Dhc and DhcA. Further, FT-IR, synchronous spectroscopy, and CD results revealed that the secondary structure of HSA was altered in the presence of Dhc and DhcA. Furthermore, the atomic force microscopy and transmission electron microscopy showed that the dimensions like height and molecular size of the HSA–Dhc and HSA–DhcA complex were larger compared to HSA alone. Detailed analysis through molecular dynamics simulations also supported greater stability of HSA–Dhc and HSA–DhcA complexes, and root-mean-square-fluctuation interpreted the binding site of Dhc as domain IB and domain IIA for DhcA. This information is valuable for further development of steroid derivative with improved pharmacological significance as novel anti-cancer drugs.

Optical and recyclable photocatalytic properties of silica supported ZnO/Au heterostructures under sun light

ZnO/Plasmonic noble metal heterostructures promise the visible light functioning photocatalysis. Here we report, silica supported ZnO/Au heterostructures for recyclable visible light photo-degradations under sun light. Firstly, silica supported ZnO nanostructures was obtained by thermal decomposition of surface adsorbed zinc acetate molecules. Further, the Au nanoparticles were formed on SiO2/ZnO in successive ionic layer adsorption and reaction process. Electron microscopic images obtained for the nanocomposite indicated the ZnO layers of ~ 30 nm and Au nanoparticles of ~ 5 nm on silica surface. Absorption spectrum of the nanocomposite indicated the bandgap value of ZnO grown on silica as 3.44 eV. Remarkably, absorption of visible region photons have got enhanced ~ 7 times for the Au nanoparticles incorporation in the silica/ZnO composite. The photocatalytic degradation of methylene blue conducted for the silica/ZnO/Au nanocomposite under sunlight have showed the 4–5 times enhanced photocatalytic effect over silica/ZnO nanocomposite. The manifestations of electron transfer from the Fermi level of Au to the ZnO facilitates the visible light functioning of the catalyst. In addition, the silica/ZnO/Au catalyst have showed the adequate efficiency in the recycled reactions.

Ultra-high resolution X-ray structures of two forms of human recombinant insulin at 100 K

The crystal structure of a commercially available form of human recombinant (HR) insulin, Insugen (I), used in the treatment of diabetes has been determined to 0.92 Å resolution using low temperature, 100 K, synchrotron X-ray data collected at 16,000 keV (λ = 0.77 Å). Refinement carried out with anisotropic displacement parameters, removal of main-chain stereochemical restraints, inclusion of H atoms in calculated positions, and 220 water molecules, converged to a final value of R = 0.1112 and Rfree = 0.1466. The structure includes what is thought to be an ordered propanol molecule (POL) only in chain D(4) and a solvated acetate molecule (ACT) coordinated to the Zn atom only in chain B(2). Possible origins and consequences of the propanol and acetate molecules are discussed. Three types of amino acid representation in the electron density are examined in detail: (i) sharp with very clearly resolved features; (ii) well resolved but clearly divided into two conformations which are well behaved in the refinement, both having high quality geometry; (iii) poor density and difficult or impossible to model. An example of type (ii) is observed for the intra-chain disulphide bridge in chain C(3) between Sγ6–Sγ11 which has two clear conformations with relative refined occupancies of 0.8 and 0.2, respectively. In contrast the corresponding S–S bridge in chain A(1) shows one clearly defined conformation. A molecular dynamics study has provided a rational explanation of this difference between chains A and C. More generally, differences in the electron density features between corresponding residues in chains A and C and chains B and D is a common observation in the Insugen (I) structure and these effects are discussed in detail. The crystal structure, also at 0.92 Å and 100 K, of a second commercially available form of human recombinant insulin, Intergen (II), deposited in the Protein Data Bank as 3W7Y which remains otherwise unpublished is compared here with the Insugen (I) structure. In the Intergen (II) structure there is no solvated propanol or acetate molecule. The electron density of Intergen (II), however, does also exhibit the three types of amino acid representations as in Insugen (I). These effects do not necessarily correspond between chains A and C or chains B and D in Intergen (II), or between corresponding residues in Insugen (I). The results of this comparison are reported.Graphical abstractConformations of PheB25 and PheD25 in three insulin structures: implications for biological activity? Insulin residues PheB25 and PheD25 are considered to be important for insulin receptor binding and changes in biological activity occur when these residues are modified. In porcine insulin and Intergen (II) PheB25 adopts conformation B and PheD25 conformation D. However, unexpectedly PheB25 in Insugen (I) human recombinant insulin adopts two distinct conformations corresponding to B and D, Figure 1 and PheD25 adopts a single conformation corresponding to B not D, Figure 2. Conformations of this residue in the ultra-high resolution structure of Insugen (I) are therefore unique within this set. Figures were produced with Biovia, Discovery Studio 2016.

Fabrication of biomass-derived C-doped Bi2WO6 templated from wood fibers and its excellent sensing of the gases containing carbonyl groups

In this work, wood fibers (Populus ussuriensis Kom) were used as biotemplate to synthesize Bi2WO6. In corporation of biomass-derived C, the sensitivity to the gases containing carbonyl groups of gas sensors is improved. The experimental results showed that the C-doped Bi2WO6 sensors presented high sensitivities to acetone, acetic acid and ethyl acetate molecules at low temperature (370°C). The gas sensitivities of C-doped Bi2WO6 sensors on exposure to 1000ppm of acetone, acetic acid and ethyl acetate gas are 4.4, 3.0 and 2.4 times higher than that exposure to 1000ppm of ethanol, respectively. That is because the biomass-derived C can adsorb more oxygen molecules and the carbonyl groups in the gases can react with more oxygen molecules and release more electrons leading a resistance decrease of the sensors due to the electronic structure of carbonyl groups. The reported strategy opened up a new way for the development of gas sensors.

Monte Carlo simulations of vapour–liquid phase equilibrium and microstructure for the system containing azeotropes

Phase equilibrium data of the mixtures including alcohols, esters and organic acids are of first interest particularly to design and optimise biodiesel production and reactive distillation processes. In this work, vapour–liquid phase equilibrium of these systems was simulated at low pressure using Gibbs ensemble Monte Carlo method. All Lennard–Jones parameters of pseudo-atoms involved in the systems were derived from previous parametrisations of TraPPE-UA force field. The Fourier coefficients of dihedrals encountered in ethyl acetate molecule have been obtained from the quantum calculations. Using this force field, temperature-composition diagrams are well reproduced for ethyl acetate + ethanol, ethyl acetate + methanol at 70.00 kPa and ethyl acetate + acetic acid mixtures at 77.33 kPa. The transferability of this force field to mixtures in these systems is noticeable. Analysis of the microstructure for the ethyl acetate + ethanol and ethyl acetate + acetic acid mixtures was presented. We found that the hydrogen bond networks consist of autoassociation and cross-association and autoassociation occupies the main position as compared with cross association in the ethyl acetate + ethanol mixture. OCHAc–HHAc and OCEtOAc–HHAc hydrogen bond interactions play a significant role in the phase behaviours or structures of ethyl acetate + acetic acid mixture.

Characterization of the High-Spin Co(II) Intermediate Species of the O2-Evolving Co4O4 Cubic Molecules.

An artificial oxygen-evolving complex (OEC) contains a tetranuclear CoIII4O4 cubic cluster ligated with acetate and pyridine molecules, a light-activated Ru(bpy)32+ moiety (bpy = 2,2'-bipyridine), and the sacrificial electron acceptor S2O82- side. A recent EPR investigation of this system showed the formation of the high-spin Co(II) ion under visible-illumination conditions. It has been supported that this center originates from the cluster and is involved in the oxygen-evolving process. The present report is focused on the further characterization of the high-spin d7 configuration of the Co(II) species. The measurement of the EPR spectrum at a wider magnetic field range in comparison to that reported recently shows the presence of an additional signal that contributes to the spectrum of the Co(II) center. The theoretical simulation of this spectrum reveals that an isotropic g value and considerably small zero-field splitting parameters describe the high-spin Co(II) ion in a unique way, which supports a tetrahedral crystal field symmetry. On the basis of the spin-Hamiltonian parameters, the looping transitions that lead to the experimental EPR signals are determined. Additionally, a possible role of the symmetry of the Co(II) species and a proposed model that explains its formation during the O2-evolving process of the Co4O4 cubic molecules are discussed.

Preferential solvation of dehydroepiandrosterone acetate in (co-solvent + ethyl acetate) mixtures according to the inverse Kirkwood–Buff integrals method

The preferential solvation parameters (δx1,3) of dehydroepiandrosterone acetate (DHEA acetate) in three co-solvent mixtures of methanol (1)+ethyl acetate (2), ethanol (1)+ethyl acetate (2), and isopropanol (1)+ethyl acetate (2) were derived from their available solubility data by means of the inverse Kirkwood–Buff integrals method. The values of δx1,3 vary non-linearly with the co-solvent (1) proportion in all the co-solvent mixtures. For the studied systems, the values of δx1,3 are negative in alcohol-rich mixtures, but positive in ethyl acetate-rich mixtures. The preferential solvation magnitude of DHEA acetate by the ethyl acetate is highest in {methanol (1)+ethyl acetate (2)} mixtures at 293.15K and 303.15K. While at 313.15K, it is highest in {isopropanol (1)+ethyl acetate (2)} mixtures. The δx1,3 values are positive indicating that DHEA acetate is preferentially solvated by ethyl acetate. The co-solvent action may be related to the disordered structure of ethyl acetate molecules around the non-polar moieties of DHEA acetate which increases the solvation having maximum values near to x1=0.25–0.35 for the three co-solvent mixtures.

Denaturation of DNA in Ternary Mixed Solution of Water/Hydrophilic/Hydrophobic Organic Solvent

Denaturation was examined for the first time in a ternary mixed solution of water/hydrophilic/ hydrophobic organic solvent using λ-DNA and a plasmid as models. The absorbance of λ-DNA and the plasmid at 260 nm gradually increased for several days up to 1.68 and 1.38 times the initial values, respectively, in a water/acetonitrile/ethyl acetate (15:3:2, volume ratio) mixed solution, whereas there was little change in a water/acetonitrile (15:3, volume ratio) mixed solution. The plasmid treated with the ternary mixed solution was also examined with agarose gel electrophoresis. These experimental data indicated that λ-DNA changed from a double helix structure to a single helix structure and that the plasmid partially transformed to generate a denaturation bubble in the structure. The new idea of using the ternary mixed solution first enabled the interaction of the hydrophobic organic solvent (e.g., ethyl acetate) molecule with the double helical structure of DNA, leading to specific slow-proceeding denaturation.

Adsorption of Ethyl Acetate from Water by Nanoporous Carbon Prepared from Waste Materials

The adsorption of ethyl acetate, a volatile organic compound, on activated carbons, synthesized from various precursors based on by-products and waste materials—polymer, biomass, coal tar pitch—was studied. The activated carbons were prepared by thermochemical treatment of the precursors, carbonization, and subsequent activation with water vapor. Surface and textural properties of obtained carbon adsorbents were characterized by low-temperature N2 adsorption, Boehm’s method, etc. The activated carbons are distinguished by relatively high surface area and developed pore structure. The adsorption investigations were performed with water solutions of ethyl acetate, and the obtained results fit well the Langmuir model, as well as the Freundlich model. All activated carbons demonstrated considerably high adsorption capacity in the range 160–450 mg/g. The obtained data indicate that the adsorption ability of activated carbon toward ethyl acetate depends on the surface area, and it increases with increasing the content of mesopores, where ethyl acetate molecules are preferably adsorbed.

In Situ ATR-FTIR Investigation of the Effects of H2o and D2o Adsorption on the TiO2 Surface

Surface hydroxyl groups on TiO2 play an important role for the photoinduced hydrophilicity as well as for photocatalytic reactions. Adsorbed species such as water and organic compounds are expected to affect the hydroxyl group behaviour on the TiO2 surface, however, their influences have to date not been clearly determined. Herein, a detailed investigation of H2O and D2O adsorption on the TiO2 surface (anatase) in the dark and under UV(A) irradiation is reported using ATR-FTIR spectroscopy. The adsorption and photocatalytic degradation of acetate on the TiO2 surface was also investigated in H2O and D2O. Adsorption of H2O with different ratios of D2O revealed an isotopic exchange reaction occurring on the surface of TiO2 in the dark. This isotopic exchange leads to a change in the behaviour of acetate with the intensity of the OH bending band between the carboxylate and the OH group at 900 cm-1 decreasing and ultimately disappearing in the dark. Upon illumination with UV(A) light both, OH and OD groups are formed, which in turn increase the hydrophilicity of the TiO2 surface. The amount of OH and OD groups was found to increase in the presence of molecular oxygen. This increase is most likely caused by the photoadsorption of H2O and D2O due to photoinduced charge transfer processes [1]. On the other hand, the investigations show that during UV(A) irradiation the acetate molecules can adsorb again on the TiO2 surface which is explained by an increased surface area due to a de-aggregation of the particles agglomerates [2]. Based upon the observed correlation between the carboxylate and the OH and/or OD groups at neutral pH (pH ≈ pHzpc) it is obvious that acetate preferentially adsorbs on the positively charged anatase in a monodentate structure. As an in-situ technique, ATR-FTIR studies provide evidence of the adsorption behaviour of water and D2O on TiO2 surfaces thus yielding new insights into the mechanism of photoinduced hydrophilicity and of photocatalytic reactions.

Three alcohol dehydrogenase genes and one acetyl-CoA synthetase gene are responsible for ethanol utilization in Yarrowia lipolytica

The non-conventional yeast Yarrowia lipolytica is able to utilize a wide range of different substrates like glucose, glycerol, ethanol, acetate, proteins and various hydrophobic molecules. Although most metabolic pathways for the utilization of these substrates have been clarified by now, it was not clear whether ethanol is oxidized by alcohol dehydrogenases or by an alternative oxidation system inside the cell.In order to detect the genes that are required for ethanol utilization in Y. lipolytica, eight alcohol dehydrogenase (ADH) genes and one alcohol oxidase gene (FAO1) have been identified and respective deletion strains were tested for their ability to metabolize ethanol. As a result of this, we found that the availability of ADH1, ADH2 or ADH3 is required for ethanol utilization in Y. lipolytica. A strain with deletions in all three genes is lacking the ability to utilize ethanol as sole carbon source. Although Adh2p showed by far the highest enzyme activity in an in vitro assay, the availability of any of the three genes was sufficient to enable a decent growth. In addition to ADH1, ADH2 and ADH3, an acetyl-CoA synthetase encoding gene (ACS1) was found to be essential for ethanol utilization.As Y. lipolytica is a non-fermenting yeast, it is neither able to grow under anaerobic conditions nor to produce ethanol. To investigate whether Y. lipolytica may produce ethanol, the key genes of alcoholic fermentation in S. cerevisiae, ScADH1 and ScPDC1, were overexpressed in an ADH and an ACS1 deletion strain. However, instead of producing ethanol, the respective strains regained the ability to use ethanol as single carbon source and were still not able to grow under anaerobic conditions.

Isomer detection on the basis of analyte adduct formation with the components of the mobile phase and tandem mass spectrometry

To investigate and compare the detection of isomers on the basis of analyte adduct formation with the components of the mobile phase appropriate infusion experiments were performed with tandem mass spectrometry (−Q1 MS, −EPI and −MRM mode) and analyses with liquid chromatography–tandem mass spectrometry (−ESI). In experiments performed following adducts were focused: deprotonated analyte adduct with sodium acetate, deprotonated analyte dimer adduct with sodium and deprotonated analyte adduct with two molecules of sodium acetate. α-hydroxybutyrate, β-hydroxybutyrate and γ-hydroxybutyrate were used as model drugs since their effective separation and detection are a real analytical challenge when γ-hydroxybutyrate is the analyte of interest. The achieved results revealed that the drugs investigated produce similar adduct ions in the negative electrospray mode. However, β-hydroxybutyrate is not a potential interfering substance for γ-hydroxybutyrate analysis other than α-hydroxybutyrate. The interference of α-hydroxybutyrate can be minimised when analyte adduct ion fragmentation is used, since appropriate fragmentations do not produce ion fragments (m/z=85) with efficiency required at different values of collision energy. Finally it was demonstrated that the strategy presented can be a real advantage when interfering substances with similar retention times do not produce adduct ions or produce adduct ions but with a different fragmentation pattern.