Thermostable lipases from the extreme thermophilic anaerobic bacteria Thermoanaerobacter thermohydrosulfuricus SOL1 and Caldanaerobacter subterraneus subsp. tengcongensis

Two novel genes encoding for heat and solvent stable lipases from strictly anaerobic extreme thermophilic bacteria Thermoanaerobacter thermohydrosulfuricus (LipTth) and Caldanaerobacter subterraneus subsp. tengcongensis (LipCst) were successfully cloned and expressed in E. coli. Recombinant proteins were purified to homogeneity by heat precipitation, hydrophobic interaction, and gel filtration chromatography. Unlike the enzymes from mesophile counterparts, enzymatic activity was measured at a broad temperature and pH range, between 40 and 90°C and between pH 6.5 and 10; the half-life of the enzymes at 75°C and pH 8.0 was 48 h. Inhibition was observed with 4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride and phenylmethylsulfonylfluorid indicating that serine and thiol groups play a role in the active site of the enzymes. Gene sequence comparisons indicated very low identity to already described lipases from mesophilic and psychrophilic microorganisms. By optimal cultivation of E. coli Tuner (DE3) cells in 2-l bioreactors, a massive production of the recombinant lipases was achieved (53–2200 U/l) Unlike known lipases, the purified robust proteins are resistant against a large number of organic solvents (up to 99%) and detergents, and show activity toward a broad range of substrates, including triacylglycerols, monoacylglycerols, esters of secondary alcohols, and p-nitrophenyl esters. Furthermore, the enzyme from T. thermohydrosulfuricus is suitable for the production of optically pure compounds since it is highly S-stereoselective toward esters of secondary alcohols. The observed E values for but-3-yn-2-ol butyrate and but-3-yn-2-ol acetate of 21 and 16, respectively, make these enzymes ideal candidates for kinetic resolution of synthetically useful compounds

Diffuse venting at the ASHES hydrothermal field : heat flux and tidally modulated flow variability derived from in situ time-series measurements

Author Posting. © American Geophysical Union, 2016. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Geochemistry, Geophysics, Geosystems 17 (2016): 1435–1453, doi:10.1002/2015GC006144.Time-series measurements of diffuse exit-fluid temperature and velocity collected with a new, deep-sea camera, and temperature measurement system, the Diffuse Effluent Measurement System (DEMS), were examined from a fracture network within the ASHES hydrothermal field located in the caldera of Axial Seamount, Juan de Fuca Ridge. The DEMS was installed using the HOV Alvin above a fracture near the Phoenix vent. The system collected 20 s of 20 Hz video imagery and 24 s of 1 Hz temperature measurements each hour between 22 July and 2 August 2014. Fluid velocities were calculated using the Diffuse Fluid Velocimetry (DFV) technique. Over the ∼12 day deployment, median upwelling rates and mean fluid temperature anomalies ranged from 0.5 to 6 cm/s and 0°C to ∼6.5°C above ambient, yielding a heat flux of 0.29 ± 0.22 MW m−2 and heat output of 3.1± 2.5 kW. Using a photo mosaic to measure fracture dimensions, the total diffuse heat output from cracks across ASHES field is estimated to be 2.05 ± 1.95 MW. Variability in temperatures and velocities are strongest at semidiurnal periods and show significant coherence with tidal height variations. These data indicate that periodic variability near Phoenix vent is modulated both by tidally controlled bottom currents and seafloor pressure, with seafloor pressures being the dominant influence. These results emphasize the importance of local permeability on diffuse hydrothermal venting at mid-ocean ridges and the need to better quantify heat flux associated with young oceanic crust.NSF Grant Numbers: OCE-1131772, OCE-1131455, OCE-1337473; University of Washington, and the NSF award Grant Number: OCE-09579382016-10-2

New Results in Four and Five Loop QED calculations

We report on two recent multiloop results in QED: (i) the four-loop corrections to the conversion relations between the QED charge renormalized in the on-shell and MS-bar schemes; (ii) analytical evaluation of a class of asymptotic contributions to the muon anomaly at five-loops.Comment: Talk presented at 9th DESY Workshop on Elementary Particle Theory: Loops and Legs in Quantum Field Theory, Sondershausen, Germany, 20-25 April 200

Sensing and Integration of Erk and PI3K Signals by Myc

The transcription factor Myc plays a central role in regulating cell-fate decisions, including proliferation, growth, and apoptosis. To maintain a normal cell physiology, it is critical that the control of Myc dynamics is precisely orchestrated. Recent studies suggest that such control of Myc can be achieved at the post-translational level via protein stability modulation. Myc is regulated by two Ras effector pathways: the extracellular signal-regulated kinase (Erk) and phosphatidylinositol 3-kinase (PI3K) pathways. To gain quantitative insight into Myc dynamics, we have developed a mathematical model to analyze post-translational regulation of Myc via sequential phosphorylation by Erk and PI3K. Our results suggest that Myc integrates Erk and PI3K signals to result in various cellular responses by differential stability control of Myc protein isoforms. Such signal integration confers a flexible dynamic range for the system output, governed by stability change. In addition, signal integration may require saturation of the input signals, leading to sensitive signal integration to the temporal features of the input signals, insensitive response to their amplitudes, and resistance to input fluctuations. We further propose that these characteristics of the protein stability control module in Myc may be commonly utilized in various cell types and classes of proteins

Consecutive reactions of aromatic?OH adducts with NO, NO₂ and O₂: benzene, toluene, m- and p-xylene, hexamethylbenzene, phenol, m-cresol and aniline

International audienceConsecutive reactions of adducts, resulting from OH radicals and aromatics, with the tropospheric scavenger molecules O2, NO and NO2 have been studied for benzene, toluene, m- and p-xylene, hexamethylbenzene, phenol, m-cresol and aniline by observing decays of OH at temperatures where the thermal back-decomposition to OH is faster than 3 s?1, typically between 300 and 340 K. The experimental technique was resonance fluorescence with flash photolysis of water as source of OH. Biexponential decays were observed in the presence of either O2 or NO, and triexponential decays were obtained in the presence of NO2. The kinetic analysis was performed by fitting the relevant rate constants of the reaction mechanism to whole sets of decays obtained at various concentrations of aromatic and scavenger. In the case of hexamethylbenzene, the biexponential decays suggest the existence of the ipso-adduct, and the slightly higher necessary temperatures show that it is even more stable. In addition, smog chamber experiments at O2 concentrations from atmospheric composition down to well below 100 ppm have been carried out for benzene, toluene and p-xylene. The drop of the effective rate constant of removal by OH occurs at reasonable O2 levels, given the FP/RF results. Comparison of the adduct reactivities shows for all aromatics of this study that the reaction with O2 predominates over that with NO2 under all tropospheric conditions, and that a reaction with NO may only occur after the reaction with O2

Some reactions of DMSO and MSIA in the aqueous phase of sea-spray: A contribution to CMD-APP and EC-project EL-CID

The reactions of DMS with ozone and MSIA and MSA with H202 and OH were studied by stopped flow, UV spectrophotometry and ion chromatography. The rapid reaction of DMS with ozone was confirmed and found to be at the limit of capabilities of the stopped-flow technique. MSA was found to be relatively inert compared with MSIA, the reaction of both compounds with H2O2 is slow enough (i.e. can be neglected in the atmosphere) to allow the use of H2O2 as OH precursor in photochemical studies of OH-induced photooxidation, leading to sulphate, finally. The pH-dependence of the UV spectra of MSIA and MSA was determined in a wide range of pH and the dissociation constants were determined