Identification of the active-site peptide of 2,3-dihydroxybenzoic acid decarboxylase from Aspergillus oryzae

The non-oxidative decarboxylation of aromatic acids is a poorly understood reaction. The transformation of 2,3-dihydroxybenzoic acid to catechol in the fungal metabolism of indole is a prototype of such a reaction. 2,3-Dihydroxybenzoic acid decarboxylase (EC 4.1.1.46) which catalyzes this reaction was purified to homogeneity from anthranilate induced cultures of Aspergillus oryzae using affinity chromatography. The enzyme did not require cofactors like NAD(+), PLP, TPP or metal ions for its activity. There was no spectral evidence for the presence of enzyme bound cofactors. The preparation, which was adjudged homogeneous by the criteria of SDS-PAGE, sedimentation analysis and N-terminal analysis, was characterized for its physicochemical and kinetic parameters. The enzyme was inactivated by group-specific modifiers like diethyl pyrocarbonate (DEPC) and N-ethylmaleimide (NEM). The kinetics of inactivation by DEPC suggested the presence of a single class of essential histidine residues, the second order rate constant of inactivation for which was 12.5 M(-1) min(-1). A single class of cysteine residues was modified by NEM with a second order rate constant of 33 M(-1) min(-1). Substrate analogues protected the enzyme against inactivation by both DEPC and NEM, suggesting the Location of the essential histidine and cysteine to be at the active site of the enzyme. The incorporation of radiolabelled NEM in a differential labelling experiment was 0.73 mol per mol subunit confirming the presence of a single essential cysteine per active-site. Differentially labelled enzyme was enzymatically cleaved and the peptide bearing the label was purified and sequenced. The active-site peptide LLGLAETCK and the N-terminal sequence MLGKIALEEAFALPRFEEKT did not bear any similarity to sequences reported in the Swiss-Prot Protein Sequence Databank, a reflection probably of the unique primary structure of this novel enzyme. The sequences reported in this study will appear in the Swiss-Prot Protein Sequence Databank under the accession number P80402.

Interaction of sheep liver apo-serine hydroxymethyltransferase with pyridoxal-5'-phosphate: A physicochemical, kinetic, and thermodynamic study

Sheep liver serine hydroxymethyltransferase (EC 2.1.2.1) is a homotetramer of M(r) 213,000 requiring pyridoxal-5'-phosphate (PLP) as cofactor, Removal of PLP from the holoenzyme converted the enzyme to the apo form which, in addition to being inactive, was devoid of the characteristic absorption spectrum. Upon the addition of PLP to the apoenzyme, complete activity was restored and the visible absorption spectrum with a maximum at 425 nm was regained. The interaction of PLP with the apoenzyme revealed two phases of reaction with pseudo-first-order rate constants of 20 +/- 5 s(-1) and 12.2 +/- 2.0 x 10(-3) s(-1), respectively. However, addition of PLP to the apoenzyme did not cause gross conformational changes as evidenced by circular dichroic and fluorescence spectroscopy. Although conformationally apoenzyme and holoenzyme were indistinguishable, they had distinct apparent melting temperatures of 51 +/- 2 and 58 +/- 2 degrees C, respectively, and the reconstituted holoenzyme was thermally as stable as the native holoenzyme. These results suggested that there was no apparent difference in the secondary structure of holoenzyme, apoenzyme, and reconstituted holoenzyme, However, sedimentation analysis of the apoenzyme revealed the presence of two peaks of S-20,S-w values of 8.7 +/- 0.5 and 5.7 +/- 0.3 S, respectively. A similar pattern was observed when the apoenzyme was chromatographed on a calibrated Sephadex G-150 column. The first peak corresponded to the tetrameric form (M(r) 200,000 +/- 15,000) while the second peak had a M(r) of 130,000 +/- 10,000. Reconstitution experiments revealed that only the tetrameric form of the apoenzyme could be converted into an active holoenzyme while the dimeric form could not be reconstituted into an active enzyme. These results demonstrate that PLP plays an important role in maintaining the structural integrity of the enzyme by preventing the dissociation of the enzyme into subunits, in addition to its function in catalysis. (C) 1996 Academic Press, Inc.

Analysis of residual tropical cemented soil behaviour

In -situ soils in gee-material spectrum might arise due to sedimentation or could be non-sedimentary residual formations. The inherent nature and diversity of geological processes involved in the soil formation stage itself are responsible for a wide variability in the in-situ state of the soil. In this paper the possibility of analyses to arrive at engineering parameters of residual soils with varied degrees of residual or acquired cementation by the use of physical and in-situ parameters normally determined in routine investigations, are examined. An Intrinsic State Line,(ISL), with reference to an intrinsic state parameter (e/e(L)) and its variation with effective stress for reconstituted clays has been developed for residual tropical soils of non-sedimentary origin. In relation to the Intrinsic State Line (ISL), the undisturbed state, e, the potential parameter, e(L), along with the overburden pressure data has been analyzed to identify the dominance of cementation or stress history or both in controlling the compressibility and strength behaviour of natural residual soil. The location of yield stress point in relation to the ISL, pre-, and post- yield stress, compression indices along the e- log sigma(v) path provide a simple means to the analysis of the compressibility characteristics of cemented soils for analysis.

Influence of Bottom Bubbling Rate on Formation of Metal Emulsion in Al-Cu Alloy and Molten Salt System

In steel refining process, an increase of interfacial area between the metal and slag through the metal droplets emulsified into the slag, so-called ``metal emulsion'', is one prevailing view for improving the reaction rate. The formation of metal emulsion was experimentally evaluated using Al-Cu alloy as metal phase and chloride salt as slag phase under the bottom bubbling condition. Samples were collected from the center of the salt phase in the container. Large number of metal droplets were separated from the salt by dissolving it into water. The number, surface area, and weight of the droplets increased with the gas flow rate and have local maximum values. The formation and sedimentation rates of metal droplets were estimated using a mathematical model. The formation rate increased with the gas flow rate and has a local maximum value as a function of gas flow rate, while the sedimentation rate is independent of the gas flow rate under the bottom bubbling condition. Three types of formation mode of metal emulsion, which occurred by the rupture of metal film around the bubble, were observed using high speed camera. During the process, an elongated column covered with metal film was observed with the increasing gas flow rate. This elongated column sometimes reached to the top surface of the salt phase. In this case, it is considered that fine droplets were not formed and in consequence, the weight of metal emulsion decreased at higher gas flow rate.

Tectono-geomorphic and environmental set-up deduced during deposition of Mio-Pleistocene sediments in NW Himalaya, India

Detailed pedofacies characterization along-with lithofacies investigations of the Mio-Pleistocene Siwalik sediments exposed in the Ramnagar sub-basin have been studied so as to elucidate variability in time and space of fluvial processes and the role of intra- and extra-basinal controls on fluvial sedimentation during the evolution of the Himalayan foreland basin (HFB). Dominance of multiple, moderately to strongly developed palaeosol assemblages during deposition of Lower Siwalik (similar to 12-10.8 Ma) sediments suggest that the HFB was marked by Upland set-up of Thomas et al. (2002). Activity of intra-basinal faults on the uplands and deposition of terminal fans at different times caused the development of multiple soils. Further, detailed pedofacies along-with lithofacies studies indicate prevalence of stable tectonic conditions and development of meandering streams with broad floodplains. However, the Middle Siwalik (similar to 10.8-4.92 Ma) sub-group is marked by multistoried sandstones and minor mudstone and mainly weakly developed palaeosols, indicating deposition by large braided rivers in the form of megafans in a Lowland set-up of Thomas et al. (2002). Significant change in nature and size of rivers from the Lower to Middle Siwalik at similar to 10 Ma is found almost throughout of the basin from Kohat Plateau (Pakistan) to Nepal because the Himalayan orogeny witnessed its greatest tectonic upheaval at this time leading to attainment of great heights by the Himalaya, intensification of the monsoon, development of large rivers systems and a high rate of sedimentation, hereby a major change from the Upland set-up to the Lowland set-up over major parts of the HFB. An interesting geomorphic environmental set-up prevailed in the Ramnagar sub-basin during deposition of the studied Upper Siwalik (similar to 4.92 to <1.68 Ma) sediments as observed from the degree of pedogenesis and the type of palaeosols. In general, the Upper Siwalik sub-group in the Ramnagar sub-basin is subdivided from bottom to top into the Purmandal sandstone (4.92-4.49 Ma), Nagrota (4.49-1.68 Ma) and Boulder Conglomerate (<1.68 Ma) formations on the basis of sedimentological characters and change in dominant lithology. Presence of mudstone, a few thin gravel beds and dominant sandstone lithology with weakly to moderately developed palaeosols in the Purmandal sandstone Fm. indicates deposition by shallow braided fluvial streams. The deposition of mudstone dominant Nagrota Fm. with moderately to some well developed palaeosols and a zone of gleyed palaeosols with laminated mudstones and thin sandstones took place in an environment marked by numerous small lakes, water-logged regions and small streams in an environment just south of the Piedmont zone, perhaps similar to what is happening presently in the Upland region/the Upper Gangetic plain. This area is locally called the `Trai region' (Pascoe, 1964). Deposition of Boulder Conglomerate Fm. took place by gravelly braided river system close to the Himalayan Ranges. Activity along the Main Boundary Fault led to progradation of these environments distal-ward and led to development of on the whole a coarsening upward sequence. (C) 2014 Elsevier B.V. All rights reserved.

Optically Stimulated Luminescence (OSL) Dating of Sediments from Himalaya

Optically Stimulated Luminescence (OSL) dating gives the age of most recent daylight exposure or heating of samples to >400 degrees C or the formation events of authigenic minerals. These correspond to the age of sedimentation and burial, ages of thermal events like contact heating by lava flows and heating during faulting and sand dyke formation, and the formation of a mineral via chemical precipitation. With the first observation of OSL in 1985, this method now occupies centre stage in Quaternary Geochronology. The use of OSL method for sediments from Himalaya began over three decades ago. The method has since provided chronology for a variety of events, such as past glaciation events, formation ages of river terraces, paleo-lacustrine deposits, landslides, floods, seismic events with substantive new insights into timing and style of geological processes. Theoretically, the dating range of method is present to a Million years, and this critically depends on two factors, viz, luminescence properties of mineral and their radiation environments. The general working range using quartz is 200ka, and using feldspars is up to Brunhes Matuyam Boundary. Extensions beyond this limit are currently being explored.

重力对异向聚集过程影响的探讨

自90年代初,分散体系聚集过程的微重力研究开始受到重视,并有一系列论文发表[1－4].Folkersma[3,4]等人在最近发表的文章中,公布了他们的最新研究结果.在探空火箭实现的微重力条件下,他们发现聚集速率要比地面实验快11倍多.为了解释这一现象,他们把重力的影响分为两部分,即沉降和自然对流.用密度匹配法,检验了在无沉降时的聚集过程.得到的结果是,聚集速率在无重力沉降影响时会比有沉降时快2.7倍.即便如此,微重力实验的聚集速率仍高出4.2倍.他们推断,这一巨大差异是由在地面上无法避免的自然对流造成的.

A study of the influence of free convection on coagulation process

It was assumed [1, 2] that gravity affects the coagulation process in two ways: free convection, which is hard to be avoided on the ground and sedimentation, which can be greatly reduced by the density-matching method. We present a ground-based experiment set-up to study the influence of convection on the perikinetic coagulation for aqueous polystyrene (PS) dispersions. The turbidity measurement was used to evaluate the relative coagulation rate and convection-driven flows in the solution were checked with a visual-magnification system. The pattern of flow field temperature profile in the sample cell is given. Our experiments show that there was no noticeable difference of coagulation rate observed no matter whether convection flows exist (with the flow speed up to 180 mu m/s) or not.

Observation Of Diffusion Process Of A Water Droplet Immersed In Protein Solution In Microgravity

Pure liquid - liquid diffusion driven by concentration gradients is hard to study in a normal gravity environment since convection and sedimentation also contribute to the mass transfer process. We employ a Mach - Zehnder interferometer to monitor the mass transfer process of a water droplet in EAFP protein solution under microgravity condition provided by the Satellite Shi Jian No 8. A series of the evolution charts of mass distribution during the diffusion process of the liquid droplet are presented and the relevant diffusion coefficient is determined.

Hacia una ética de la responsabilidad: Derrida y el otro “por venir” en Lévinas

Resumen: La tesis del trabajo es que la deconstrucción entraña en sí misma una actitud ética, y que ésta radica intrínsecamente en su apertura al otro y en la responsabilidad frente a ese otro. Para ello, se intenta en primer lugar una elucidación del concepto, caracterizándola como un protocolo de lectura compuesto de dos fases: des-sedimentación y reinscripción, siendo este último momento el que la identifica como una lectura singular y no universalizable. Posteriormente, a través de las nociones de “sujeto” y “alteridad del texto”, se muestra cómo las bases de la ética deconstructiva giran en torno a las nociones de “yo” y “otro”, conceptos que Derrida, como se demuestra, toma de Lévinas. La instancia ética es la apertura al otro, que se verifica por la responsabilidad que supone concebir al otro como aquello incapaz de apropiación. Dicha responsabilidad configura la aporía ética de lo general-particular, al concebirse como la posibilidad imposible de una opción general que es al mismo tiempo una decisión personal o singular, o, en otros términos, la indecidibilidad entre la norma general y la decisión concreta, que es a su vez la tensión entre “justicia”, “hospitalidad” y “negociación”. Lo anterior demuestra que el acto ético (o acto deconstructivo) tiene la misma estructura de la différance, imbricada siempre en la iteración del significante.

The Fluid Phenomena In the Crystallization Of the Protein Crystal

This paper reports that an optical diagnostic system consisting of Mach-Zehnder interferometer with a phase shift device and image processor has been used for study of the kinetics of protein crystal growing process. The crystallization process of protein crystal by vapour diffusion is investigated. The interference fringes are observed in real time. The present experiment demonstrates that the diffusion and the sedimentation influence the crystallization of protein crystal which grows in solution, and the concentration capillary convection associated with surface tension occurs at the vicinity of free surface of the protein mother liquor, and directly affects on the outcome of protein crystallization. So far the detailed analysis and the important role of the fluid phenomena in protein crystallization have been discussed a little in both space- and ground-based crystal growth experiments. It is also found that these fluid phenomena affect the outcome of protein crystallization, regular growth, and crystal quality. This may explain the fact that many results of space-based investigation do not show overall improvement.

Study on improving the turbidity measurement of the absolute coagulation rate constant

The existing theories dealing with the evaluation of the absolute coagulation rate constant by turbidity measurement were experimentally tested for different particle-sized (radius = a) suspensions at incident wavelengths (lambda) ranging from near-infrared to ultraviolet light. When the size parameter alpha = 2 pi a/lambda > 3, the rate constant data from previous theories for fixed-sized particles show significant inconsistencies at different light wavelengths. We attribute this problem to the imperfection of these theories in describing the light scattering from doublets through their evaluation of the extinction cross section. The evaluations of the rate constants by all previous theories become untenable as the size parameter increases and therefore hampers the applicable range of the turbidity measurement. By using the T-matrix method, we present a robust solution for evaluating the extinction cross section of doublets formed in the aggregation. Our experiments show that this new approach is effective in extending the applicability range of the turbidity methodology and increasing measurement accuracy.

沉降对不同粒径聚苯乙烯胶乳球异向聚集的影响

R.Folkersma等报道了在微重力环境下2μm聚苯乙烯（PS）胶乳球的异向聚集速率有明显增大的结果,本文作者之一孙祉伟等的实验结果却与此有很大差异.为此作者在孙祉伟等的实验基础上对1,2,3μm以及1μm+2μm PS胶乳球混和体系的相对聚集速率进行了研究.作者对原有实验装置进行了改进,并验证了改进后的实验装置的可行性.用密度匹配法实现模拟微重力条件,用快聚集过程中浊度随时间的变化表示相对聚集速率.结果表明,重力引起的沉降对所研究体系聚集速率的影响是很小的,在实验误差范围内可以忽略.作者认为与Stein等结果显著不同的原因是二者使用的样品表面性质不同以及实验方法的差异.

Slope Stability Analysis Using Anisotropic and Nonlinear Failure Criteria

The shear strength of soils or rocks developed in a landslide usually exhibits anisotropic and nonlinear behavior. The process of sedimentation and subsequent consolidation can cause anisotropy of sedimentary soils or rocks, for instance. Nonlinearity of failure envelope could be attributed to "interlocking" or "dilatancy" of the material, which is generally dependent upon the stress level. An analytical method considering both anisotropy and nonlinearity of the failure envelops of soil and rocks is presented in the paper. The nonlinearfailure envelopes can be determined from routine triaxial tests. A spreadsheet program, which uses the Janbu's Generalized Procedure of Slice and incorporates anisotropic, illustrates the implementation of the approach and nonlinearfailure envelops. In the analysis, an equivalent Mohr-Coulomb linear failure criterion is obtained by drawing a tangent to the nonlinear envelope of an anisotropic soil at an appropriate stress level. An illustrative example is presented to show the feasibility and numerical efficiency of the method.

Submarine landslides : selected studies in the U.S. Exclusive Economic Zone

Submarine Landslides: An Introduction 1 By RIo Lee, W.C. Schwab, and J.S. Booth U.S. Atlantic Continental Slope Landslides: Their Distribution, General Anributes, and Implications 14 By J.S. Booth, D.W. O'Leary, Peter Popenoe, and W.W. Danforth Submarine Mass Movement, a Formative Process of Passive Continental Margins: The Munson-Nygren Landslide Complex and the Southeast New England Landslide Complex 23 By D.W. O'Leary The Cape Fear Landslide: Slope Failure Associated with Salt Diapirism and Gas Hydrate Decomposition 40 By Peter Popenoe, E.A. Schmuck, and W.P. Dillon Ancient Crustal Fractures Control the Location and Size of Collapsed Blocks at the Blake Escarpment, East of Florida 54 By W.P. Dillon, J.S. Risch, K.M. Scanlon, P.C. Valentine, and Q.J. Huggett Tectonic and Stratigraphic Control on a Giant Submarine Slope Failure: Puerto Rico Insular Slope 60 By W.C. Schwab, W.W. Danforth, and K.M. Scanlon Slope Failure of Carbonate Sediment on the West Florida Slope 69 By D.C. Twichell, P.C. Valentine, and L.M. Parson Slope Failures in an Area of High Sedimentation Rate: Offshore Mississippi River Delta 79 By J.M. Coleman, D.B. Prior, L.E. Garrison, and H.J. Lee Salt Tectonics and Slope Failure in an Area of Salt Domes in the Northwestern Gulf of Mexico 92 By B.A. McGregor, R.G. Rothwell, N.H. Kenyon, and D.C. Twichell Slope Stability in Regions 01 Sea-Floor Gas Hydrate: Beaufort Sea Continental Slope 97 By R.E. Kayen and H.J. Lee Mass Movement Related to Large Submarine Canyons Along the Beringian Margin, Alaska 104 By P.R. Carlson, H.A. Karl, B.D. Edwards, J.V. Gardner, and R. Hall Comparison of Tectonic and Stratigraphic Control of Submarine Landslides on the Kodiak Upper Continental Slope, Alaska 117 By M.A. Hampton Submarine Landslides That Had a Significant Impact on Man and His Activities: Seward and Valdez, Alaska 123 By M.A. Hampton, R.W. Lemke, and H.W. Coulter Processes Controlling the Style of Mass Movement in Glaciomarine Sediment: Northeastern Gulf of Alaska 135 By W.C. Schwab and H.J. Lee Contents V VI Contents Liquefaction of Continental Shelf Sediment: The Northern California Earthquake of 1980 143 By M.E. Field A Submarine Landslide Associated with Shallow Sea-Floor Gas and Gas Hydrates off Northern California 151 By M.E. Field and J.H. Barber, Jr. Sur Submarine Landslide, a Deep-Water Sediment Slope Failure 158 By C.E. Gutmacher and W.R. Normark Seismically Induced Mudflow in Santa Barbara Basin, California 167 By B.D. Edwards, H.J. Lee, and M.E. Field Submarine Landslides in a Basin and Ridge Setting, Southern California 176 By M.E. Field and B.D. Edwards Giant Volcano-Related Landslides and the Development of the Hawaiian Islands 184 By W.R. Normark, J.G. Moore, and M.E. Torresan Submarine Slope Failures Initiated by Hurricane Iwa, Kahe Point, Oahu, Hawaii 197 By W.R. Normark, Pat Wilde, J.F. Campbell, T.E. Chase, and Bruce Tsutsui (PDF contains 210 pages)