Kinetics of thermochemical pretreatment of lignocellulosic materials

The results of an experimental study of the acid hydrolysis of hardwood are presented in the form of values for the three parameters, activation energy, power on the acid concentration, and pre-exponen-tial factor, of the first order kinetic constants for each of the following reaction participants: xylan remaining, glucan remaining, xylose formed, and xylose decomposed. These are used as a base for a quantitative theory to predict the temperature, time, and acid concentrations needed for effective pretreatment of the substrate for subsequent enzymatic hydrolysis of the glucan. This theory is based on the assumption that successful pretreatment requires >90% removal of the xylan, <10% removal of the glucan, and >80% xylose yield. This theory is compared with selected published data.     

Overproduction of riboflavin by anArthrobacter sp. mutant resistant to 5-fluorouracil

Effects of pH and dissolved oxygen concentration on batchwise riboflavin production by a 5-fluorouracil (5-FU)-resistant mutant ofArthrobacter sp. were investigated. The reaction was carried out in a jar fermentor. The optimal pH of culture medium was around 7.3. Dissolved oxygen concentration was almost constant during fermentation at 600 rpm of agitation rate. Production of riboflavin reached a maximum of 160 mg/L after 70 h fermentation under the agitation rate of 600 rpm, aeration rate of 1.0 L/min, and pH 7.0.     

Mechanism of crystallization of enzyme protein thermolysin

The mechanism of crystallization of enzyme protein thermolysin was investigated. The size distribution of thermolysin precipitates was measured by dynamic light scattering during precipitation, and the surface and cross section of the finally obtained precipitate were observed by scanning electron microscopy. The thermolysin precipitates obtained at the initial supersaturation ratio of 8.9 to 164 and pH 7.0 were hexagonal rods having an average size of 9.2×1.5 μm, and were composed of a number of small particles of 15 to 200 nm in diameter. The average size of the small particles was 60 nm in diameter, and the formation of the particles was found to be completed in the early stage of precipitation. Observation of the finally recovered thermolysin precipitate by polarizing microscopy revealed that the precipitate is a crystalline solid. From these data, a possible mechanism of thermolysin crystallization was proposed. The crystallization proceeds through two steps: the first step is the formation of primary particles, and the second step is crystal growth by highly ordered aggregation of the primary particles.     

Surface area of pretreated lignocellulosics as a function of the extent of enzymatic hydrolysis

Partially purified S1 nuclease was bound through its carbohydrate moiety to Con A-Sepharose containing increasing amounts of lectin. The retention of activity was high, varying essentially from 75% on the "low lectin" matrix (1 mg Con A/mL of Sepharose), to no detectable activity on the "high lectin" matrix (8 mg Con A/mL of Sepharose). However, approximately 50% activity could be restored in "high lectin" matrix when the coupling was carried out in the presence of glucose, suggesting that the loss of activity on the "high lectin" matrix is caused by conformational changes brought about by the multiple attachment of the enzyme to the matrix. Interaction of Con A with S1 nuclease was used to predict the nature of carbohydrate moiety and its location with respect to the active site of the enzyme. Immobilization resulted in an increase in the optimum temperature, pH, and temperature stabilities, but it did not affect the pH optimum. A marginal increase in the apparent Km was observed. The bound enzyme also showed enhanced stability toward 8 M urea. On repeated use, the bound enzyme retained more than 80% of its initial activity after 6 cycles. These results are discussed taking into consideration the factors affecting immobilized enzymes. In addition, the potential use of immobilized S1 nuclease as an analytical tool is discussed.     

An extension of the Harano-Ooshima rate expression for enzymatic hydrolysis of cellulose to account for changes in the amount of adsorbed cellulase

Three empirical rate expressions, Kinetics I, II, and III, for the enzymatic hydrolysis of cellulose were evaluated in an effort to develop a easy-to-use rate expression. They are based on the following equation:-dV/dX = kV, where V and X are the hydrolysis rate and the fractional conversion. In Kinetic I,k is constant. In Kinetic II, a linear relatinship betweenk and t is assumed. In Kinetic III, an exponential relationship is assumed. The three expressions were applied to enzymatic hydrolysis carried out under seven different conditions in which the kinds of substrates, enzymes, and initial concentrations were varied. All of the examined rate expressions were applied to the hydrolysis with success, but the better accuracies were obtained by Kinetic III, Kinetic II, and Kinetic I in this order. The variations ofk with time found in this study, especially the exponential relationship, were consistent with the effect of the measured changes in the concentration of adsorbed enzyme as predicted by theory developed previously by Ooshima et al. (1).     

Kinetics of enzymatic hydrolysis of lignocellulosic materials based on surface area of cellulose accessible to enzyme and enzyme adsorption on lignin and cellulose

Comparison of the model with experimental data is currently in progress. It appears that more detailed studies of the adsorption dynamics, not just adsorption equilibrium, are needed.     

Removal of inhibition by ammonium ion in nitrogenase-dependent hydrogen evolution of a marine photosynthetic bacterium,Rhodopseudomonas sp. strain W-1S

Applied Biochemistry and Biotechnology - The addition of NH4Cl at concentrations of more than 1 mM completely inhibited nitrogenase-dependent hydrogen evolution using 1 mM succinate as a substrate...     

Correlation study among oxygen permeability,molecular mobility,and amorphous structure change of poly(ethylene‐vinylalcohol copolymers) by moisture

Oxygen permeability and the free‐volume hole size (cavity size) of ethylene‐vinylalcohol copolymers (EVOH) indicate abnormal humidity dependence, that is, they have minimum values in the range of around 20–40% RH, not showing a monotonic increase with humidity. To clarify this abnormal phenomenon, we investigated its molecular mobility and amorphous structure change by means of solid‐state NMR and temperature‐modulated differential scanning calorimetry (TMDSC). The glass transition temperature (T_g) decreased with humidity. Specimens stored at 15–60% RH showed large enthalpy relaxation, and it was found that the amorphous structure became more compact and the molecular conformation became more stable by ageing within this range of humidity. Under these conditions, solid‐state NMR measurement showed a component with intermediate relaxation time in the amorphous region. The results obtained by TMDSC and solid‐state NMR showed a reduction in molecular mobility by densifying in the amorphous region under the condition of 15–60% RH. The fact that the oxygen permeability and the cavity size of EVOH indicate minimum values at low humidity are attributed to the reduction in molecular mobility by enthalpy relaxation. © 2009 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 47: 1181–1191, 2009     

Creep deformation of single crystals of new Co–Al–W-based alloys with fcc/L12 two-phase microstructures

The creep deformation behaviour of single crystals of Co–Al–W-based alloys with γ?+?γ′ two-phase microstructures has been investigated in tension under a constant stress of 137?MPa in air at 1000°C as a function of the γ′ solvus temperature and the volume fraction of the γ′ phase. When described by the creep strain rate versus time curve, the creep deformation of Co–Al–W-based alloys consists of transition and accelerating regions without a steady-state region, as observed in many modern nickel-based alloys. However, the creep strength of the present Co–Al–W-based alloys is comparable with nickel-based superalloys of the first generation but is much weaker than those of the second and higher generations. Unlike in nickel-based superalloys, the so-called p (parallel)-type raft structure, in which the γ′ phase is elongated along the tensile axis direction, is formed during creep in Co–Al–W-based alloys, being consistent with what is expected from the positive values of lattice misfit between the γ and γ′ phases. As a result, of the alloys investigated, the best creep properties are obtained with the alloy possessing the highest volume fraction (85%) of the γ′ phase, which is far larger than usual for nickel-based superalloys (55–60%).