Balancing the effect of pretreatment severity on hemicellulose extraction and pulping performance during auto-hydrolysis prior to kraft pulping of acacia wood

When using a combination of pre-extraction and chemical pulping, a high yield of sugar recovery and minimal negative effect on the subsequent pulping step are expected. In this work, the P factor was utilized to investigate the effect of auto-hydrolysis severity on sugar recovery, removal of the main component, and impact on the kraft pulping of acacia wood chips. Using a P factor of 235, 84.34% of the polysaccharides in 14.05 g L⁻¹ of dissolved sugars could be obtained. In addition, the soluble sugars were easily separated with a recovery yield of 3.54 g ·L⁻¹ and Mw of 4,690 g mol⁻¹ by direct precipitation using organic solvents. However, a maximum of 22.14 g L⁻¹ of dissolved sugars was obtained with approximately 72.53% polysaccharides and Mw of 2,198 g mol⁻¹ for a P factor of 601. Moreover, nearly 50% of the degraded carbohydrates remained in the auto-hydrolyzed wood chips. The decrease in the mass of pentosan, holocellulose, and klason lignin was 62, 30, and 8.76%, respectively. With intensifying severity, the screened yield and viscosity of pulps decreased markedly, whileas the Kappa number increased. No significant differences were observed in the morphology of the resultant fibers. Moreover, there was a decrease in the physical strength of the pulps due to the loss of the intrinsic strength of the pulp fibers, which in turn resulted from the cellulose damage. The combustion performance of the resultant pulping black liquor is improved due to the higher lignin content.     

Structure of hardwood glucuronoxylans: modifications and impact on pulp retention during wood kraft pulping

The behaviour of different hardwood glucuronoxylans during the kraft pulping process was investigated. Woods and pulps xylans were isolated and characterized by size exclusion chromatography, methylation (linkage) analysis and ¹H NMR. Eucalyptus globulus and Eucalyptus urograndis showed xylan retention significantly higher than that of Betula pendula. The higher retention of Eucalyptus xylans was assigned to (i) their higher average molecular weight (31 against 24 KDa in B. pendula) and to (ii) the presence of O-2 substituted 4-O-methyl--d-glucuronic acid groups ([→2)-GlcpA-(1→]) with galactopyranosyl/glucopyranosyl residues belonging to fragments of galactan/glucan chains that were absent in B. pendula xylans. A significant part of uronic acids, particularly [→2)-GlcpA-(1→] units, remain in fibres until the end of pulping. The acetylation degree and distribution of acetyl groups between Xylp units, in general terms, was similar in the three types of xylans. Unexpectedly, about 20% of the acetyl groups persisted in pulps xylans till the end of pulping.     

Extracting value from Eucalyptus wood before kraft pulping: effects of hemicelluloses solubilization on pulp properties

Eucalyptus globulus wood samples were subjected to autohydrolysis for extracting hemicelluloses, and the resulting solids were assayed as substrates for kraft pulping and further Totally Chlorine Free (TCF) bleaching. The susceptibility of treated solids to kraft processing was assessed under selected experiments covering the optimum experimental range. In order to establish a basis for comparison, samples of untreated wood were also subjected to kraft delignification. The best kraft pulps obtained from autohydrolyzed solids were subjected to an optimized TCF bleaching sequence involving double alkaline oxygen and pressurized H₂O₂ processing, and characterized using standard methods. The suitability of the final product obtained by autohydrolysis-kraft delignification-TCF bleaching for specific purposes is discussed on the basis of the experimental results.     

A kinetic model for plasmid curing

A simple mathematical model of drug-induced plasmid elimination (curing) considering density-dependent growth rates and plasmid transfers is presented. It describes nonlinear population dynamics of conjugative plasmids during in vitro curing experiments in batch culture. The model was tested on kinetics of acridine orange curing of F'lac plasmid. Effects of density dependence, plasmid elimination, selection for plasmidless segregants, conjugation, initial and maximal population density, and postsegregational killing on curing kinetics are simulated and discussed.     

A kinetic model for calcium distribution

Models for the distribution of minerals in the body are of interest as they allow researchers to trace the effect of a dose on mineral levels in plasma, storage and other compartments. Limited models are available in the literature for tracing the distribution of a calcium dose through a short time period. We propose a more general kinetic model which includes both limited absorption through the gut and loss of calcium via excretion. This new method has the advantages of giving reasonable results over moderate time periods, and allowing the extrapolation of calcium levels in extracellular fluid and storage. We fit the model to published data in order to obtain typical parameter values. These values are then used to analyze the implications of the model regarding the effect of calcium dose on calcium levels in various compartments.     

A kinetic model for cell agglutination

A model of concanavalin A (ConA) mediated cell agglutination kinetics is proposed, in which the binding of the lectin, the agglutination of cells and the disintegration of cell clumps are discussed. This resulted in a differential equation, which is solved in terms of the average number of cells per cell clump as a function of time.     

A model for oscillatory kinetic systems

A kinetic model is proposed for oscillatory kinetic phenomena. The exact analytic solution is exhibited and shown to account for several features exhibited by oscillatory chemical and biological systems.     

A kinetic model for glutamate dehydrogenase

A model for the glutamate dehydrogenase reaction has been obtained that contains the reported intermediates suggested by binding and equilibrium isotope exchange methods. Calculated steady state-initial velocity rates using this model are quantitatively consistent with a wide range of nonlinear experimental data in both directions.     

A kinetic model for the quantitative analysis of complement

A simple model has been developed which accurately predicts the time course of complement mediated lysis of sensitized red cells. The model assumes that the one hit theory of immune hemolysis is applicable and that the rate of lysis is directly proportional to the concentration of a complement component present in rate limiting amounts. It also assumes that the rate of lysis is dependent on the fraction of cells lysed. The model can be related to the classical von Krogh equation for end point complement analyses and can be used to estimate the rate constant for the critical step in hemolysis, as well as the efficiency of the critical complement component in the rate limiting step. Parameters derived from the model can be quantitatively related to complement concentration and can be used as the basis for a quantitative assay of complement activity. The model can also be used to calculate, for a particular sample, the concentration at which complement activity becomes undectable, the complement activity of the pure, undiluted sample, and the time required for the sample to produce complete lysis of the available cells.     

A vesicle capture sensor chip for kinetic analysis of interactions with membrane-bound receptors

A novel sensor chip for use in surface plasmon resonance (SPR) biosensors has been developed to capture vesicles which may contain membrane-bound receptors. Sulforhodamine-containing vesicles were shown by fluorescence microscopy to be immobilized intact on the sensor chip. Binding of cholera toxin to captured vesicles containing ganglioside GM(1) was demonstrated using SPR, and the derived kinetic and affinity constants were similar to literature values. Biotinylated vesicles captured on the sensor chip were used to bind streptavidin and then biotinylated ss-DNA. The hybridization of complementary ss-DNA to the immobilized ss-DNA was then analyzed using SPR. The values obtained were similar to those obtained for an identical interaction analyzed using a commercially available streptavidin-containing sensor chip. Binding of vancomycin-group antibiotics to captured vesicles containing a bacterial cell wall mucopeptide analogue was demonstrated. No binding of the bacterial endotoxin Cry1A(c) to captured vesicles containing its cell surface receptor could be demonstrated.     

A kinetic model for microbial growth on solid hydrocarbons

A kinetic model has been presented to explain the growth of microorganism on solid hydrocarbons. The model is based on the assumption that metabolite produced by the growing cells helps the dissolution of the solid substrate in the aqueous medium. The linear behavior of the growth curve predicted by the model is verified experimentally.     

A kinetic model for microbial growth on liquid hydrocarbons

A kinetic model is presented to explain microbial growth using liquid n-alkanes as substrate. The model is based on the assumption that growth occurs on the soluble alkane and that the metabolite produced by the growing cells helps the dissolution of liquid alkanes in the aqueous medium. Growth curves based on that model fit well with growth data for batch and continuous culture reported by various authors. The model also explains the differences between the relative length of exponential and linear phases of growth reported earlier.     

A kinetic model for subtractive hybridization.

Nucleic acid sequences that differ in abundance between two populations (target sequences) can be cloned by multiple rounds of subtractive hybridization and amplification by PCR. These sequences can be cDNAs representing up-regulated mRNAs, or genomic DNAs from deletion mutants. We have derived an equation that describes the recovery of such sequences, and have used this to simulate the outcome of up to 10 rounds of subtractive hybridization and PCR amplification. When the model was tested by comparing its predictions with the published results from genomic and cDNA subtractions, the predictions of the model were generally in good agreement with the published data. We have modelled the outcomes of genomic subtractions, for a variety of genomes, and have used it to compare various strategies for enriching targets. The model predicts that for genomes of less than 5 x 10(8) bp, deletions of as small as 1 kbp should represent > 99% of the DNA after three to six rounds of hybridization (depending on the enrichment procedure). As genomes increase in size, the kinetics of hybridization become an important limiting factor. However, even for genomes as large as 3 x 10(9) bp, it should be possible to isolate deletions of 5 kbp using the appropriate conditions. These simulations suggest that such methods offer a realistic alternative to chromosome walking for identifying genomic deletions for which there are known phenotypes, thereby considerably reducing time and effort. For cDNA subtractive hybridization, the model predicts that after six rounds of hybridization, sequences that do not differ in abundance between the tester and driver populations (the background) will represent < 1% of the subtracted population, and even quite modestly upregulated cDNAs should be successfully enriched. Where several up-regulated cDNAs are present, the predicted final representation is dependent on both the initial abundance and the degree of up-regulation.     

A kinetic model for quantitative evaluation of the effect of hydrogen and osmolarity on hydrogen production by Caldicellulosiruptor saccharolyticus

Background

The main technological impediment to widespread utilization of lignocellulose for the production of fuels and chemicals is the lack of low-cost technologies to overcome its recalcitrance. Organisms that hydrolyze lignocellulose and produce a valuable product such as ethanol at a high rate and titer could significantly reduce the costs of biomass conversion technologies, and will allow separate conversion steps to be combined in a consolidated bioprocess (CBP). Development of Saccharomyces cerevisiae for CBP requires the high level secretion of cellulases, particularly cellobiohydrolases.

Results

We expressed various cellobiohydrolases to identify enzymes that were efficiently secreted by S. cerevisiae. For enhanced cellulose hydrolysis, we engineered bimodular derivatives of a well secreted enzyme that naturally lacks the carbohydrate-binding module, and constructed strains expressing combinations of cbh1 and cbh2 genes. Though there was significant variability in the enzyme levels produced, up to approximately 0.3 g/L CBH1 and approximately 1 g/L CBH2 could be produced in high cell density fermentations. Furthermore, we could show activation of the unfolded protein response as a result of cellobiohydrolase production. Finally, we report fermentation of microcrystalline cellulose (Avicel?) to ethanol by CBH-producing S. cerevisiae strains with the addition of beta-glucosidase.

Conclusions

Gene or protein specific features and compatibility with the host are important for efficient cellobiohydrolase secretion in yeast. The present work demonstrated that production of both CBH1 and CBH2 could be improved to levels where the barrier to CBH sufficiency in the hydrolysis of cellulose was overcome.     

A generalized kinetic model for the study of microbial growth

The following general equation is proposed to represent the kinetics of microbial growth \documentclass{article}\pagestyle{empty}\begin{document}$$\phi (dR/dt) + \psi R + X = 0$$\end{document}, where phi and psi depend on several parameters of the fermenting system. The values of phi and psi were calculated based on results obtained in a batch lactic acid fermentation, a batch cultivation of yeast on diesel oil, and a continuous cultivation of yeast on sugarcane molasses.     

A kinetic model for the burst phase of processive cellulases

Cellobiohydrolases (exocellulases) hydrolyze cellulose processively, i.e. by sequential cleaving of soluble sugars from one end of a cellulose strand. Their activity generally shows an initial burst, followed by a pronounced slowdown, even when substrate is abundant and product accumulation is negligible. Here, we propose an explicit kinetic model for this behavior, which uses classical burst phase theory as the starting point. The model is tested against calorimetric measurements of the activity of the cellobiohydrolase Cel7A from Trichoderma reesei on amorphous cellulose. A simple version of the model, which can be solved analytically, shows that the burst and slowdown can be explained by the relative rates of the sequential reactions in the hydrolysis process and the occurrence of obstacles for the processive movement along the cellulose strand. More specifically, the maximum enzyme activity reflects a balance between a rapid processive movement, on the one hand, and a slow release of enzyme which is stalled by obstacles, on the other. This model only partially accounts for the experimental data, and we therefore also test a modified version that takes into account random enzyme inactivation. This approach generally accounts well for the initial time course (approximately 1 h) of the hydrolysis. We suggest that the models will be useful in attempts to rationalize the initial kinetics of processive cellulases, and demonstrate their application to some open questions, including the effect of repeated enzyme dosages and the 'double exponential decay' in the rate of cellulolysis.