An Enhanced Process for the Production of a Highly Purified Extracellular Lipase in the Non-conventional Yeast <Emphasis Type="Italic">Yarrowia lipolytica</Emphasis>

Yarrowia lipolytica LgX64.81 is a non-genetically modified mutant that was previously identified as a promising microorganism for extracellular lipase production. In this work, the development of a fed-batch process for the production of this enzyme in this strain was described. A lipolytic activity of 2,145 U/mL was obtained after 32 h of batch culture in a defined medium supplemented with 10 g/L of tryptone, an enhancer of lipase expression. To maximize the volumetric productivity, two different fed-batch strategies had been investigated. In comparison to batch process, the intermittent fed-batch strategy had not improved the volumetric lipase productivity. In contrast, the stepwise feeding strategy combined with uncoupled cell growth and lipase production phases resulted in a 2-fold increase in the volumetric lipase productivity, namely, the lipase activity reached 10,000 U/mL after 80 h of culture. Furthermore, this lipase was purified to homogeneity by anion exchange chromatography on MonoQ resin followed by gel filtration on Sephacryl S-100. This process resulted in an overall yield of 72% and a 3.5-fold increase of the specific lipase activity. The developed process offers a great potential for an economic production of Lip2 at large scale in Y. lipolytica LgX64.81.     

First emission studies of Tc2X8(2-) systems (X = Cl, Br)

The emission spectra of the solids [n-Bu(4)N](2)Tc(2)X(8) (X = Cl, Br) have been investigated at room temperature and 77 K. In each case, the emission originates in the (1)δ-δ* excited state, as with the rhenium homologues, but has a shorter lifetime.     

Synthesis of a new hydrophilic poly(ethylene glycol)-ionic liquid and its application in peptide synthesis

The synthesis of a new hydrophilic ammonium-based poly(ethylene glycol)-ionic liquid (PEG-IL) is reported; the structure was assigned by NMR ((1)H, (13)C) and MALDI mass spectrometry. The viscosity and thermal stability were also studied, as well as its polarity. Its application as an alternative solvent in the synthesis of dipeptides under microwave irradiation is also described.     

An automated instrument for human STR identification: design, characterization, and experimental validation

The microfluidic integration of an entire DNA analysis workflow on a fully integrated miniaturized instrument is reported using lab‐on‐a‐chip automation to perform DNA fingerprinting compatible with CODIS standard relevant to the forensic community. The instrument aims to improve the cost, duration, and ease of use to perform a “sample‐to‐profile” analysis with no need for human intervention. The present publication describes the operation of the three major components of the system: the electronic control components, the microfluidic cartridge and CE microchip, and the optical excitation/detection module. Experimental details are given to characterize the level of performance, stability, reliability, accuracy, and sensitivity of the prototype system. A typical temperature profile from a PCR amplification process and an electropherogram of a commercial size standard (GeneScan 500?, Applied Biosystems) separation are shown to assess the relevance of the instrument to forensic applications. Finally, we present a profile from an automated integrated run where lysed cells from a buccal swab were introduced in the system and no further human intervention was required to complete the analysis.     

Study on the bubble transport mechanism in an acoustic standing wave field

The use of bubbles in applications such as surface chemistry, drug delivery, and ultrasonic cleaning etc. has been enormously popular in the past two decades. It has been recognized that acoustically-driven bubbles can be used to disturb the flow field near a boundary in order to accelerate physical or chemical reactions on the surface. The interactions between bubbles and a surface have been studied experimentally and analytically. However, most of the investigations focused on violently oscillating bubbles (also known as cavitation bubble), less attention has been given to understand the interactions between moderately oscillating bubbles and a boundary. Moreover, cavitation bubbles were normally generated in situ by a high intensity laser beam, little experimental work has been carried out to study the translational trajectory of a moderately oscillating bubble in an acoustic field and subsequent interactions with the surface. This paper describes the design of an ultrasonic test cell and explores the mechanism of bubble manipulation within the test cell. The test cell consists of a transducer, a liquid medium and a glass backing plate. The acoustic field within the multi-layered stack was designed in such a way that it was effectively one dimensional. This was then successfully simulated by a one dimensional network model. The model can accurately predict the impedance of the test cell as well as the mode shape (distribution of particle velocity and stress/pressure field) within the whole assembly. The mode shape of the stack was designed so that bubbles can be pushed from their injection point onto a backing glass plate. Bubble radial oscillation was simulated by a modified Keller–Miksis equation and bubble translational motion was derived from an equation obtained by applying Newton’s second law to a bubble in a liquid medium. Results indicated that the bubble trajectory depends on the acoustic pressure amplitude and initial bubble size: an increase of pressure amplitude or a decrease of bubble size forces bubbles larger than their resonant size to arrive at the target plate at lower heights, while the trajectories of smaller bubbles are less influenced by these factors. The test cell is also suitable for testing the effects of drag force on the bubble motion and for studying the bubble behavior near a surface.     

Local hydrodynamic investigation of the aeration in a submerged hollow fibre membranes cassette

The better understanding of the effective air distribution inside a membrane cassette is a particular challenge in submerged membrane bioreactor. The present study is the first one that investigates the hydrodynamics of the coarse bubbles flow inside a hollow fibre membranes cassette. The experimental investigations were carried out in a reactor equipped with commercial modules from ZENON ZeeWeed^® 500d. A bi-optical probe was used to measure the bubble size, the bubble velocity and the gas hold-up at different locations between the modules and for three different gas flow rates. These local measurements gave significant information about the lateral distribution of the air and its evolution with the height on the surface of the membrane modules, which can impact on the filtration performance and are the first step to an optimisation of the aeration system and module geometry.     

Graphes et ordonnés démontables, propriété de la clique fixe

This work extends to dismantable graphs many properties of dismantable posets dealing with products, exponentiation, existence of paths in a graph of all homomorphisms, fixed clique property, etc. We show that a poset is dismantlable in the sense of Rival if and only if its comparability graph is dismantlable, thus proving that, for posets, dismantlability is a comparability invariant. We establish the analogues for graphs of results of Duffus, Poguntke and Rival about fixed point sets and cores in posets.     

Co-continuity interval in immiscible polymer blends by dynamic mechanical spectroscopy in the molten and solid state

The work focuses on the detection of the co-continuity window in immiscible polymer blends. The purpose of the paper is to describe how rheological techniques can help to evaluate the composition range of the co-continuous morphology through the study of a particular system: PEO/PVDF-HFP. First, the blends were characterized by selective dissolution experiments and SEM observations. Then the ability of dynamic mechanical spectroscopy to detect the co-continuity was investigated in the melt and in the solid state. The evolution of the storage modulus of molten blends with their composition at a constant low frequency gives information about the co-continuity interval, especially as far as the onset of the continuity of the PEO phase is concerned. Then the immiscibility of the polymers and the continuity of PVDF-HFP as a function of blend composition have been highlighted by means of dynamic mechanical spectrometry below the melting point of PVDF-HFP. Comparison with results from classical methods shows fair agreement.This paper was presented at the first Annual European Rheology Conference (AERC) held in Guimarães, Portugal, September 11-13, 2003.     

The cantilever strip plate under torision, bending or flexure at infinity

A homogeneous, isotropic plate occupies the region 0x ₁, |x ₂|a, |x ₃|h, where the ratio h/a is sufficiently small so that the classical theory of thin plate bending applies. The short end of the plate at x ₁=0 is clamped while the long sides are free. This cantilever plate is now loaded at x ₁=+ by an applied twisting moment, by a bending moment or by flexure. Despite the fundamental nature of these problems, and the long history of thin plate theory, no solutions are to be found in the existing literature that will determine (for instance) the important unknown resultants V ₁, M ₁₁ at the clamped end x ₁=0. The main reason for this is that this combination of boundary conditions leads to severe oscillating singularities of the field in the corners (0, ±a). The fact that such singularities must exist is widely known, but we present here for the first time a method of solution that takes these singularities fully into account.Our numerical results show that the values of M ₁₁, V ₁ on x ₁=0 bear little resemblance to those of the corresponding Saint-Venant solutions, which do not fully satisfy the boundary conditions at the clamped end. Indeed, significantly large values of these resultants were found at points far enough from the corners so as to be relevant in actual engineering applications. Also of interest are certain weighted integrals of M ₁₁, V ₁ which we calculate. These constants determine the effect of the clamping at large distances (greater than 4a, say) from the cla,ped end. At such distances, the effect of the clamping is merely to impose an additional rigid body deflection on the plate.Finally, we consider the plate of finite length. Provided that the aspect ratio is 2 or more, we give accurate approximate solutions for the torsion, bending or flexure of a finite plate clamped at both ends.