Effect of physical pretreatment on dilute acid hydrolysis of water hyacinth (Eichhornia crassipes)

Effects of different physical pretreatments on water hyacinth for dilute acid hydrolysis process (121 ± 3 °C, 5% H₂SO₄, 60 min) were comparatively investigated. Untreated sample had produced 24.69 mg sugar/g dry matter. Steaming (121 ± 3 °C) and boiling (100 ± 3 °C) for 30 min had provided 35.9% and 52.4% higher sugar yield than untreated sample, respectively. The highest sugar yield (132.96 mg sugar/g dry matter) in ultrasonication was obtained at 20 min irradiation using 100% power. The highest sugar production (155.13 mg sugar/g dry matter) was obtained from pulverized samples. Hydrolysis time was reduced when using samples pretreated by drying, mechanical comminution and ultrasonication. In most methods, prolonging the pretreatment period was ineffective and led to sugar degradations. Morphology inspection and thermal analysis had provided evidences of structure disruption that led to higher sugar recovery in hydrolysis process.     

Experimental studies for levulinic acid production from whole kernel grain sorghum.

Levulinic acid has potential as an important basic chemical material. This study proposed a method of making levulinic acid using abundant and low cost whole kernel sorghum grain as the raw material. Flour made from grinding whole kernel sorghum grains was blended with 2%, 5% and 8% aqueous solutions of sulfuric acid. Mixtures were heated to 160 or 200 degrees C in a pressurized reactor. A stepwise heating scheme helped improve the yield of levulinic acid. Levulinic acid yield was determined based on sorghum flour content, as opposed to total sorghum mass. Levulinic acid yield increased as reaction temperature increased. Higher sulfuric acid concentration also significantly increased the levulinic acid yield. However, flour loading had an adverse effect on levulinic acid yield. A maximum yield of 32.6% levulinic acid was achieved at 200 degrees C, 8% sulfuric acid concentration and 10% flour loading. A linear regression model was capable of predicting the levulinic acid yield with respect to effects of reaction temperature, mineral acid concentration and flour loading (R2 = 0.88).     

The mechanism of the acid-catalysed hydrolysis of polysaccharides

The kinetics of homogeneous and heterogeneous acid-catalyzed hydrolysis of polysaccharides has been studied. The hydrolysis of O-methylcellulose in solutions of hydrochloric, sulfuric, and perchloric acids, and the acetolysis of cellulose triacetate were found to follow the mechanism established earlier for glycosides. Degradation of O-ethylcellulose films in hydrochloric acid vapour occurred in the kinetic region.     

Identification of hexose hydrolysis products in metabolic flux analytes: a case study of levulinic acid in plant protein hydrolysate

Biosynthetically directed fractional ¹³C labeling, a popular methodology of metabolic flux analysis, involves culture on a mixture of ¹³C and ¹²C substrates and preparation a ‘metabolic flux analyte’ (typically protein hydrolysate) from the biomass. Metabolic flux analytes prepared from complex eukaryotes may contain additional compounds than those prepared from microorganisms. We report the presence of such compounds (hexose hydrolysis products) in a plant metabolic flux analyte (acid hydrolyzed protein from soybean embryos). We designed NMR experiments to systematically identify these compounds, and found that they were levulinic acid (LVA; major) and hydroxyacetone (HyA; minor). These acid hydrolysis products of hexoses (glucose and mannose) were generated during acid hydrolysis of glycosylating sugars (glucosamine and mannose) associated with soybean embryo protein. Analysis of LVA by two-dimensional [¹³C, ¹H] NMR and measurement of its J-coupling constants revealed long-range coupling between atoms C3 and C5, which enables LVA to provide more isotopomer information than its precursor hexose. Furthermore, we found that LVA and HyA preserve the isotopomeric composition of the metabolic hexose from which they are derived. An important consequence of these results is that comparison of LVA and HyA isotopomers from two separate metabolic flux analytes (protein hydrolysate and starch hydrolysate) from the same plant tissue can distinguish between parallel glycolysis and pentose phosphate pathways in different subcellular compartments.     

Application of modelling techniques to the planning of in vitro arsenic kinetic studies

A kinetic model describing the hepatic methylation of arsenite [As(III)] was developed on the basis of limited data from in vitro mechanistic studies. The model structure is as follows: sequential enzymic methylation of arsenite to its monomethylated (MMA) and dimethylated (DMA) products by first-order and Michaelis-Menten kinetics, respectively; uncompetitive inhibition of the formation of DMA by As(III); and first-order reversible binding of As(III), MMA and DMA to cytosolic proteins. Numerical sensitivity analysis was used to evaluate systematically the impact of changes in input parameters on model responses. Sensitivity analysis was used to investigate the possibility of designing experiments for robust testing of the uncompetitive inhibition hypothesis, and for further refining the model. Based on the sensitivity analysis, the MMA concentration is the most important response on which to focus. The parameters V(max) and k(i) can be reliably estimated by using the same concentration time-course data at intermediate initial arsenite concentrations of 1--5microM at 30 +/- 5 minutes. K(m) must be estimated independently of V(max), since the two parameters are highly correlated at all times, and the optimal experimental conditions would include lower initial concentrations of arsenite (0.1--0.5microM) and earlier time-points (about 8--18 minutes). The use of initial arsenite concentrations much above 5microM would not yield additional useful information, because the sensitivity coefficients for MMA, protein-bound MMA, DMA and protein-bound DMA tend to become extremely small or exhibit erratic trends. Overall trends in the sensitivity analysis indicated the desirability of performing measurements at times shorter than 60 minutes. This work demonstrates that physiological modelling and sensitivity analysis can be efficient tools for experimental planning and hypothesis testing when applied in the earliest phases of kinetic model development, thus allowing more-efficient and more-directed experimentation, and minimising the use of laboratory animals.     

Comparative kinetic studies on the neutral protease and thermolysin catalyzed hydrolysis of simple dipeptide substrates

A comparative study of the Bacillus subtilis neutral protease and Bacillus thermoproteolyticus thermolysin calalyzed hydrolysis of a few dipeptide sustrates including furylacryloylglycyl-L -leucine amide is reported. While differences in the k_cat/K_m were observed between the two enzymes toward substrates in which phenylalanine or leucine donated the amino group of the peptide bond, secondary effects of substituents on the carbonyl donating amino acid and pH profiles were quite similar. Differences were also observed toward protein substrates as compared to dipeptides.     

脱落酸调节植物抵御水分胁迫的机制研究

干旱、盐渍、低温等均可导致植物可利用水分的亏缺,表现为水分胁迫。植物感受到水分胁迫,诱导脱落酸（abscisic acid,ABA）生物合成。ABA可通过促使气孔关闭或抑制气孔开放,使作物尽可能地降低蒸腾失水,以抵御水分胁迫。该文就植物激素ABA及其下游信号过氧化氢（hydrogenperoxide,H2O2）、一氧化氮（nitric oxide,NO）以及Ca2＋等在植物气孔运动调节方面的研究进展进行概述,以构建水分胁迫下ABA调节植物气孔运动的可能模式。     

Distribution of cytokinin-like activity in different plant parts of the water hyacinth, Eichhornia crassipes

Cytokinin-like activity in extracts of leaf laminae, petioles, shoots, roots and flowers of young plants of the water hyacinth, Eichhornia crassipes S. was analyzed following Sephadex LH-20 column chromatography using the soybean callus bioassay. In all plant parts analyzed, two prominent peaks of cytokinin activity having elution volumes similar to zeatin and zeatin riboside were detected. Putative cytokinin gluco-side-like activity was detected only in leaves and flowers. The cytokinin complements of the leaves and the roots were qualitatively different. It would appear that cytokinins supplied by the roots are metabolized in the leaves or certain cytokinins are synthesized in the leaves themselves. The possible significance and distribution of cytokinins in different plant parts in relation to roots is discussed.     

Experimental evidence rejects pairwise modelling approach to coexistence in plant communities

Competition is often invoked as the cause of plant species loss with increasing system productivity. Experimental results for multispecies assemblages are virtually absent and mathematical models are thus used to explore the relationship between competition and coexistence. Modelling approaches to coexistence and diversity in competitive communities commonly employ Lotka-Volterra-type (LV) models with additive pairwise competitive effects.Using pairwise plant competition experiments, we calibrate the LV system and use it to predict plant biomass and coexistence in six three-species and one seven-species experimental mixture. Our results show that five out of the six three-species sets and the seven-species set deviate significantly from LV model predictions. Fitting an additional non-additive competition coefficient resulted in predictions that more closely matched the experimental results, with stable coexistence suggested in all but one case. These results are discussed with particular reference to the possible underlying mechanisms of coexistence in our experimental community. Modelling the effect of competition intensity on stability indicates that if non-additive effects occur, they will be relevant over a wide range of community sizes. Our findings caution against relying on coexistence predictions based on LV models.     

Kinetic studies on the acid hydrolysis of the methyl ketoside of unsubstituted and O-acetylated N-acetylneuraminic acid

The hydrolysis of the model compound 2-O-methyl-4,7,8,9-tetra-O-acetyl-N-acetyl-alpha-d-neuraminic acid and neuraminidase (Vibrio cholerae) closely resembled that of the O-acetylated sialic acid residues of rabbit Tamm-Horsfall glycoprotein. This confirmed that O-acetylation was responsible for the unusually slow rate of acid hydrolysis of O-acetylated sialic acid residues observed in rabbit Tamm-Horsfall glycoprotein and their resistance to hydrolysis by neuraminidase. The first-order rate constant of hydrolysis of 2-methyl-N-acetyl-alpha-d-neuraminic acid by 0.05m-H(2)SO(4) was 56-fold greater than that of 2-O-methyl-4,7,8,9-tetra-O-acetyl-N-acetyl -alpha-d-neuraminic acid. Kinetic studies have shown that in the pH range 1.00-3.30, the observed rate of hydrolysis of 2-methyl-N-acetyl-alpha-d-neuraminic acid can be attributed to acid-catalysed hydrolysis of the negatively charged CO(2) (-) form of the methyl ketoside.     

Experimental and modelling studies on the DNA cleavage by elsamicin A.

The ability of elsamicin A, an antitumour antibiotic, to cleave DNA in the presence of ferrous iron and reducing agents, has been analysed using experimental and theoretical approaches. Experimentally, the antibiotic causes DNA breakage in the presence of ferrous ions and a reducing agent. The DNA-cleaving activity appears to be partially blocked by the action of superoxide dismutase and catalase. These results indicate that the elsamicin aglycone moiety (chartarin) can be involved in the production of free radicals. We have performed a broad theoretical study based in the quantum-mechanical framework, which allow us to determine the redox properties of elsamicin that lead to the generation of radical species. Our results clearly show that elsamicin acts as a true catalyst in the production of superoxide radicals. Moreover, it is suggested that the oxidation/reduction mechanism of the aglycone moiety of elsamicin (a lactone), leading to DNA breakage, is different from the mechanism followed by other well-known anti-cancer drugs, whose chromophore is a quinone.     

Enzyme kinetic and molecular modelling studies of sulphur-containing substrates of phenylalanine 4-monooxygenase

Previous investigations into the binding of substrates/cofactors to the PAH active site have only concentrated on Phe, thienylalanine and BH₄. This is the first reported investigation to model aliphatic thioether amino acid substrates to PAH. The clearance of the thioether substrates (4.82-79.09% of Phe) in the rat and human (1.19-37.41% of Phe) showed species differences. The xenobiotic thioether substrates (SMC and SCMC) were predicted to be poor substrates for PAH by the molecular modelling investigation and this has now been confirmed by the in vitro enzyme kinetic data. However, reaction phenotyping investigations have found that PAH was the major enzyme involved in the metabolism of SCMC in vitro and in vivo.     

Experimental DNA-binding and computer modelling studies on an analogue of the anti-tumor drug amsacrine

The DNA-binding properties of the anti-cancer drug amsacrine and a 9-aminoacridine analogue substituted at the 4 position with a 4-methanesulphonanilido-group, have been examined by means of unwinding, melting and equilibrium binding experiments. These find that the latter compound is at least as effective as a DNA-binder and intercalator as amsacrine itself. Molecular modelling and energetic calculations have confirmed this, and have produced plausible intercalation geometries. These show that there are subtle differences in the low-energy minor groove arrangements adopted by the substituents of the two drugs. Speculation is advanced that these differences may be relevant to the marked differences in cytotoxicity shown by the two compounds.     

Nutrient limitation to the decomposition of water hyacinth ( Eichhornia crassipes)

The responses of decomposition to N and P supply were investigated in three leaf types of water hyacinth (Eichhornia crassipes (Mart.) Solms): dead green leaves collected from Donghu Lake; green, and brown leaves collected from outdoor tanks. The ratios of C:N, C:P, lignin:N and lignin:P were lowest in the green leaves collected from Donghu Lake, and highest in the brown leaves collected from outdoor tanks. Decomposition constant (k) of water hyacinth varied greatly, ranged from 0.006 to 0.099 d^–1. Leaf litters decayed most quickly within the initial two weeks during the experimental period, but decomposition rate decreased significantly in the following days. Decomposition and nutrient (N and P) release were fastest in the green leaves collected from Donghu Lake, intermediate in the green leaves collected from outdoor tanks, slowest in the brown leaves collected from outdoor tanks. Statistical analyses revealed that the effects of P-availability on decomposition rate and N, P release rate of the three litter types were significant, whereas the impacts of N-availability was insignificant (p > 0.05) except for the brown leaves collected from outdoor tanks. These results suggest that decomposition rate and nutrient content dynamics of water hyacinth differ with their growth habitats, and could partly be regulated by nutrient availability, especially by P-availability, in the environments.     

Kinetics of the pyrolytic and hydrothermal decomposition of water hyacinth

The kinetics of water hyacinth decomposition using pyrolysis and hydrothermal treatment was compared. With pyrolysis, initial vaporization occurred at 453 K as determined by thermogravimetric analysis, while initial solubilisation occurred at 433 K with subcritical hydrothermal treatment. The “kinetic triplet” was determined for the ranges of 423-483 K (range I) and 473-553 K (range II) using the Coats-Redfern method for both treatments. The calculated activation energies for ranges I and II were 110 and 116 kJ/mol for conventional pyrolysis and 145 and 90 kJ/mol for hydrothermal treatment. The similar activation energies for the two temperature ranges observed for pyrolysis implied that only hemicellulose decomposition occurred. For hydrothermal treatment, both hemicellulose and cellulose decomposition occurred in temperature range II, in which a notable lower activation energy was observed. This implied hydrothermal treatment was more suitable for conversion lignocellulosic biomass under these conditions.     

Pretreatment and enzymic saccharification of water hyacinth cellulose

Water hyacinth was pretreated, under variable conditions, with NaOH, alkaline H₂O₂, peracetic acid and sodium chlorite. Combined pretreatments included sodium chlorite with each of NaOH, alkaline H₂O₂ and peracetic acid. Combined pretreatment with 0.1% NaClO₂ for 1 h at 100 °C and peracetic acid at 100 °C for 15 min afforded the most promising sample. The recovered lignin, cellulose and hemicellulose of this sample was 2.56%, 96.69%, and 81.38%, respectively. The same sample, by cellulase hydrolysis showed the highest cellulose conversion (80.8%) and 90% saccharification using 200 FPU/g substrate. Some ambient factors affecting saccharification of pretreated water hyacinth were investigated. Enzymic saccharification after 6 h was about 50% of that at 48 h, indicating a slow hydrolysis rate by time. Addition of 8% glucose at the beginning of the enzymic hydrolysis decreased the saccharification to about its half while addition of 8% ethanol brought about complete inhibition of the enzyme. Addition of cellobiase to the reaction mixture increased cellulose conversion and saccharification by 10%.     

Synthesis, modelling and kinetic assays of potent inhibitors of purple acid phosphatase

Purple acid phosphatases (PAPs) are binuclear metallohydrolases that have been isolated from various mammals, plants, fungi and bacteria. In mammals PAP activity is associated with bone resorption and can lead to bone metabolic disorders such as osteoporosis; thus human PAP is an attractive target to develop anti-osteoporotic drugs. Based on a previous lead compound and rational drug design, acyl derivatives of α-aminonaphthylmethylphosphonic acid were synthesised and tested as PAP inhibitors. Kinetic analysis showed that they are good PAP inhibitors whose potencies improve with increasing acyl chain length. Maximum potency is reached when the number of carbons in the acyl chain is between 12 and 14. The most potent inhibitor of red kidney bean PAP is the dodecyl-derivative with K_ic = 5 μM, while the most potent pig PAP inhibitor is the tetradecyl-derivative with K_ic = 8 μM, the most potent inhibitor of a mammalian PAP yet reported.     

Experimental and kinetic studies on methylene blue adsorption by coir pith carbon

Varying the parameters such as agitation time, dye concentration, adsorbent dose, pH and temperature carried out the potential feasibility of thermally activated coir pith carbon prepared from coconut husk for removal of methylene blue. Greater percentage of dye was removed with decrease in the initial concentration of dye and increase in amount of adsorbent used. Kinetic study showed that the adsorption of dye on coir pith carbon was a gradual process. Lagergren first-order, second-order, intra particle diffusion model and Bangham were used to fit the experimental data. Equilibrium isotherms were analysed by Langmuir, Freundlich, Dubnin-Radushkevich, and Tempkin isotherm. The adsorption capacity was found to be 5.87 mg/g by Langmuir isotherm for the particle size 250-500 microm. The equilibrium time was found to be 30 and 60 min for 10 and 20 mg/L and 100 min for 30, 40 mg/L dye concentrations, respectively. A maximum removal of 97% was obtained at natural pH 6.9 for an adsorbent dose of 100 mg/50 mL and 100% removal was obtained for an adsorbent dose of 600 mg/50 mL of 10 mg/L dye concentration. The pH effect and desorption studies suggest that chemisorption might be the major mode of the adsorption process. The change in entropy (DeltaS0) and heat of adsorption (DeltaH0) of coir pith carbon was estimated as 117.20 J/mol/K and 30.88 kJ/mol, respectively. The high negative value of change in Gibbs free energy indicates the feasible and spontaneous adsorption of methylene blue on coir pith carbon.