A comparison of the cellulolytic activities of rumen ciliate protozoa grown under different conditions

Cell-free extracts of rumen Entodiniomorphid protozoa were prepared by gentle sonication (under conditions such that the associated bacteria were not disrupted) and centrifuged (5500g, 30 min) to produce a clear, bacteria-free supernatant fluid. These preparations contained enzymes that were active against a number of cellulose preparations. They released dye from cellulose azure, produced cellobiose from or loss of turbidity of phosphoric acid-reprecipitated cellulose, produced reducing material from and loss of viscosity of carboxymethylcellulose, and produced reducing material from microcrystalline cellulose. Unfortunately with all methods the relationship between product formed and enzyme concentration was a curve with steadily decreasing slope and activities were always expressed in terms of concentration of an extract from the rumen ciliate,Epidinium ecaudatum caudatum, a series of concentrations of which were always run in parallel with other samples. Results have been calculated as specific activities relative to that of rumen-grownEpi. ec. caudatum taken as 100. Comparison has been made, where possible, between protozoa grown as a single species in the rumen of a sheep (ration 1000g hay and 100g oats/d) (referred to as R-grown), protozoa grown in culture on dried grass alone (CG-grown) and protozoa grown in culture on dried grass and ground wheat (CGW-growri) (Coleman, 1978). WithEudiplodinium maggii, the highest activity (106–135%) was obtained with CG-grown, 23–65% with R-grown, and 5–8% with CGW-grown protozoa. WithEremoplastron bovis the values were 41–49% with R-grown, 26–43% with CG-grown and 16–23% with CGW-grown protozoa. WithOstracodinium obtusum bilobum the activities were 90–210% with R-grown, 92–134% with CG-grown, and 85–110% with CGW-grown.Ophryoscolex caudatus would not grow under condition CG but had activities of 54–125% under the other two conditions.Diploplastron affine also would not grow well under condition CG, but had 20–40% activity when R-grown and 100–110% when CGW-grown. These results show that there is no consistent pattern of enzyme activities under the growth conditions used and that there may be important species differences.     

A study on the inhibition of cellulolytic activities by lignin

It is well known that cellulolytic enzymes hardly attack lignocellulose. Hitherto knowledge, however, has not yet fully elucidated the mechanisms of the inhibition; whether it depends on a steric hindrance or protein-binding inhibition. For the purpose of acceleration of enzymatic digestion of lignocellulose biomass, the study was carried out using partially purified cellulase of Meicelase and milled wood lignin from soft or hard wood. Milled wood lignin was prepared from soft wood,Picea jezoensis, or from hard wood,Quercus serrata, by the method of Björkman. Lignin was dissolved in methyl cellosolve, since the solvent has little effect on cellulolytic activities, and enzymic activities were followed with partially purified Meicelase. The results show that cellobiose activities of enzymes fromTrichoderma viride are markedly depressed by both lignin of soft and hard wood, and the former lignin inhibits the reaction stronger than the latter. Lignins show no inhibition in the reaction with CMC and debris of filter paper. Liberation of glucose from filter paper by the enzyme in the presence of lignin is also depressed, and addition of cellobiase instead of lignin in the above enzyme reaction shows the delay of time for breakdown of filter paper. This phenomenon indicates that lignin inhibits enzymic decomposition of cellobiose in the degradation pathway of cellulose and accumulated cellobiose gives a feedback function to digestion of fibers of paper.     

GC-MS analysis of diaminopimelic acid stereoisomers and amino acid enantiomers in rumen bacteria

The amounts and the configuration of the stereoisomers of 2,6-diaminopimelic acid (Dap) and the enantiomeric content of other amino acids were determined in five individual species (Fibrobacter succinogenes, Streptococcus bovis, Selenomonas ruminantium, Prevotella ruminicola and Anaerovibrio lipolytica) of rumen bacteria, and in samples of mixed rumen bacteria isolated from sheep. The separation and quantification of the Dap stereoisomers was achieved by gas chromatography (GC) of trifluoroacetyl 2-propyl esters on a Chirasil-L-Val fused silica column, and detection was achieved by selected ion monitoring mass spectrometry (SIM-MS). No isomers of Dap were detected in S. bovis and P. ruminicola, two of the bacterial isolates. LL- and DD-Dap were not detected in any of the bacterial samples. As only the meso-isomer of Dap was detected in these microorganisms, it was quantified by adding LL-Dap as an internal standard before the bacteria were acid-hydrolyzed. Amounts of between 4.8 and 12.0 mg meso-Dap per gram of bacterial dry matter (DM) were determined. The presence in the rumen bacteria of free amino acid enantiomers, extractable with 70% aqueous ethanol, were determined by GC-SIM-MS; the D-amino acids were predominantly Ala, Asp and Glu, but there was considerable variation between the species.     

Microcalorimetric study of the biodegradation of cellulolytic compounds

Microcalorimetry can be used as a tool for the analysis of microbial growth on soluble or insoluble substrates. Modern apparatus allows studies under aerobic or anaerobic growth culture conditions. The heat quantity evolved during any microbial growth is dependent on catabolic and anabolic reactions according to:△H _met=(1-α)△H _cat+α△H _an In this relationship, δH_met, δH_cat, and δH_an are the enthalpy variations associated with metabolic, catabolic, and anabolic reactions, respectively, and a represents the molar fraction of the energy source that is incorporated into cellular carbon. The value of α is strongly influenced by culture conditions. Under anaerobic conditions, the α coefficient is low and does not exceed 0.2. Under aerobic conditions, α can reach a value as high as 0.6. Some experimental results were presented and discussed. Modern microcalorimeters record the power evolved by microbial growth as a function of time. The curves obtained are called power-time curves. These curves can be used for the determination of growth parameters. However, power-time curves allow visualization of some catabolic phenomena that are not related to growth because they occur during the stationary growth phase. Some examples were shown. More recently, microcalorimetry has been used for studies of degradation of insoluble substrates, such as lignocellulosic compounds, by pure or mixed cultures of microbes. The results obtained with pure cultures of bacteria growing on cellulose were interpreted using the same relationships that have been used for studies on soluble sugars. It was possible to show the influence of the structure of the cellulose on the kinetics of its degradation. The results obtained with mixed cultures were difficult to interprete at the present time. The power-time curves are very complex, showing many heat peaks that cannot be related to any known physiological phenomena. However, the total heat evolved during lignocellulosic fermentation by mixed cultures is proportional to the quantity of sugar fermented; this was demonstrated and discussed. Thus it can be concluded that microcalorimetry can be used for the estimation of the biodegradability of any solid substrate. This technique can also be used for evaluation of the efficiency of the pretreatments of lignocellulosic compounds that are done to increase the biodegradability of these substrates. Recent data will be shown.     

Synthesis, structure, spectra and redox activities of model compounds for methanogenic bacteria

Synthetically accessible benzimidazoles have been synthesized and the benzimidazole ligands were complexed with nickel(II) nitrate. A nickel(II) complex of N,N-bis(benzimidazole-2-ylethyl)ethylenediamine was crystallized in single-crystal form and the structure was investigated by X-ray crystallography. The structure of the complex is bicapped axial coordinated octahedral. Ni(bbes)²⁺₂[bbes=bis(benzimidazole-2-ylethyl)sulfide] exhibits broad low energy bands in electronic spectra and high redox potential in cyclic voltammetry (CV) rather than Ni(enbzim)²⁺ [enbzim =N,N-bis(benzimidazol-2-ylethyl)ethylenediamine], where high energy well separated bands were observed in the visible region and a more negative redox potential was detected in CV. Experimental studies show that an increasing amount of -orbital interaction with the Ni²⁺ion, irrespective of chelate ring may favour the higher redox potential. The higher redox potential of methanogenic bacterial [Ni(II)/Ni(I)] than nickel compounds is one of the main factors for the degradation of organic biodegradable compounds and the further transformation to methane.     

Synthesis,structure, spectra and redox activities of model compounds for methanogenic bacteria

Synthetically accessible benzimidazoles have been synthesized and the benzimidazole ligands were complexed with nickel(II) nitrate. A nickel(II) complex of N,N'-bis(benzimidazole-2-ylethyl)ethylenediamine was crystallized in single-crystal form and the structure was investigated by X-ray crystallography. The structure of the complex is bicapped axial coordinated octahedral. Ni(bbes)(2+)(2)[bbes=bis(benzimidazole-2-ylethyl)sulfide] exhibits broad low energy bands in electronic spectra and high redox potential in cyclic voltammetry (CV) rather than Ni(enbzim)(2+) [enbzim =N,N'-bis(benzimidazol-2-ylethyl)ethylenediamine], where high energy well separated bands were observed in the visible region and a more negative redox potential was detected in CV. Experimental studies show that an increasing amount of pi-orbital interaction with the Ni(2+)ion, irrespective of chelate ring may favour the higher redox potential. The higher redox potential of methanogenic bacterial [Ni(II)/Ni(I)] than nickel compounds is one of the main factors for the degradation of organic biodegradable compounds and the further transformation to methane.     

Glycosylation of the Arg-gingipains of Porphyromonas gingivalis and comparison with glycoconjugate structure and synthesis in other bacteria

Post-translational modification of proteins by covalent attachment of sugars to the protein backbone (protein glycosylation) is the most common post-translational modification in the eucaryotic cell. However, the addition of carbohydrates to proteins of Eubacteria and Archaea has been demonstrated and accepted only recently. There is now a rapidly expanding list of bacterial glycoproteins that have been characterised from a variety of different organisms including many important pathogens. The Arg-gingipains of Porphyromonas gingivalis are recent additions to this list. In this review we present a summary of our investigations on the structure of the glycan additions to these proteolytic enzymes, the genetics of the glycosylation process and some of the effects on enzyme function and recognition. These findings are placed in the context of the current status of understanding of glycoconjugate structure and synthesis in other bacteria. Given the importance of glycosylation of eucaryotic proteins to their stability, structure, resistance to proteolysis and recognition, the modifications to the proteases described in the present report are likely to have a functional role in the properties of these enzymes in periodontal disease.     

Distribution of glycoside hydrolases and polysaccharide depolymerases associated with the particulate fraction from the bovine rumen and an artificial rumen

The structural carbohydrates of plant cell walls are degraded and utilized in ruminants by the synergistic action of a number of species of organisms. Electron microscopy has shown that several different morphological groups of bacteria are attracted to the plant cell wall during the degradation process. Many of these bacteria, which act in close proximity to the cell wall, bind to the wallvia their own extracellular polymeric adhesives. In addition to the cell wall-associated organisms, two other subpopulations have been identified as those colonizing the free rumen fluid and those associated with the rumen epithelium (Cheng et al., 1979). Of the subpopulations acting on digesta, most work has been carried out on the nonassociated group, even though 50–70% of the rumen bacteria are associated with the feed particles. In this report, we show the distribution in activity of a wide variety of carbohydrate-degrading enzymes associated with the particulate fraction in both in vivo and in vitro systems. The in vivo method employed was to suspend the cell wall material in nylon bags of fixed pore size in the rumen of a cow fed on a similar diet, while the in vitro system was the artificial rumen technique (Czerkawski and Breckenridge, 1977) using rumen liquor from a sheep fed on a similar diet. The digesta and the associated liquid phase were subjected to different fractionation procedures and the activities of a wide variety of glycoside hydrolases and polysaccharide depolymerases were determined using conventional techniques. In general, the results from the in vivo and in vitro methods were in good agreement. Highest levels of activity for the various enzymes that are required for cell wall degradation were found in the populations attached to or closely associated with the feed particles. Included in these enzymes were cellulases, hemicellulases, Β-D-xylopyranosidase, and α-D-arabinofuranosidase. Separate functional groups of organisms could be recognized in the particle-associated population by their distinctive profiles. The activity of enzymes not associated with cell wall degradation (e.g., amylases and α-d-glucosidase) were low, although high levels were found in the free rumen fluid.     

Screening genus Penicillium for producers of cellulolytic and xylanolytic enzymes

For enzymatic hydrolysis of lignocellulosic material, cellulolytic enzymes from Trichoderma reesei are most commently used, but, there is a need for more efficient enzyme cocktails. In this study, the production of cellulolytic and xylanolytic enzymes was investigated in 12 filamento us fungi from genus Penicillium and compared with that of T. reesei. Either Solka-Floc cellulose or oat spelt xylan was used as carbon source in shake flask cultivations. All the fungi investigated showed coinduction of cellulolytic and xylanolytic enzymes during growth on cellulose as well as on xylan. The highest filter paper activity was measured after cultivation of Penicillium brasilianum IBT 20888 on cellulose.     

A new green method for the synthesis of silver nanoparticles and their antibacterial activities against gram‐positive and gram‐negative bacteria

This study aims to evaluate the capability of Ageratum conyzoides and Mikania micrantha extracts to synthesize silver nanoparticles (AgNPs) and their antibacterial capability against gram‐positive and gram‐negative bacteria. Several properties of the synthesized AgNPs, including plasmonic, biomolecule bonding, shape, size, and antibacterial, were investigated. Ultraviolet–visible (UV–vis) spectroscopy was employed for characterizing their plasmonic properties. Functional groups on the produced AgNPs were investigated by Fourier‐transform infrared (FT‐IR) spectroscopy. The size and shape of the AgNPs were identified using the field‐emission scanning electron microscopy (FESEM). Inhibition zone measurement was carried out for evaluating the antibacterial capability. This study showed that the extracts of A. conyzoides and M. micrantha were able reducing agents as evidenced by the formation of the spherical AgNPs. UV–vis spectroscopy, FT‐IR spectroscopy, and FESEM confirmed the physicochemical characteristics of AgNPs. AgNPs that were synthesized using M. micrantha were slightly smaller than those produced using A. conyzoides. In general, the present work establishes that the synthesized AgNPs have antibacterial capability depending on their size and synthesis procedure.     

Effect of monascus pigment derivatives on the electrophoretic mobility of bacteria, and the cell adsorption and antibacterial activities of pigments

Various amino acid derivatives of monascus pigments were synthesized. The effects of pigment derivatives on the pigment adsorption ratio, electrophoretic mobility (EPM) of bacterial cells, and antibacterial activity were investigated under varying conditions of pigment type, pigment concentration, pH, and ionic strength. Two hydrophobic and two hydrophilic derivatives were selected as model pigments. There was a close relationship between the antimicrobial activity and the pigment adsorption ratio. Against Escherichia coli, the hydrophobic l-Tyr and l-Phe derivatives (log P = 3.18 and 3.57) exhibited high antimicrobial activities (MIC = 8 and 16 mg/L) and high cellular adsorption ratios (9.6 and 10.9 mg/L). The hydrophilic l-Glu and l-Asn derivatives (log P = 1.40 and 0.47) exhibited low activities (MIC = 64 and 128 mg/L) and low adsorption ratios (4.7 and 4.0 mg/L). The electrophoretic mobility of 11 different bacteria varied between −1.93 × 10⁻⁸ and −1.19 × 10⁻⁸ m² V⁻¹ s⁻¹ regardless of Gram⁺ or Gram⁻. The l-Phe derivative showed low MIC values (high antimicrobial activities) against bacteria with a high electrophoretic mobility. A positive linearity between the pigment adsorption ratio and the electrophoretic mobility was established. When the four pigment derivatives were added to E. coli solutions, the electrophoretic mobility of cells in all cases sharply increased with an increasing pigment concentration. The mobility value was high for hydrophobic pigment derivatives in descending order of l-Phe (0.8 × 10⁻⁸ m² V⁻¹ s⁻¹), l-Tyr (0.68 × 10⁻⁸ m² V⁻¹ s⁻¹), l-Glu (0.46 × 10⁻⁸ m² V⁻¹ s⁻¹), and l-Asn (0.44 × 10⁻⁸ m² V⁻¹ s⁻¹). Additional adsorption of the hydrophobic derivatives probably occurred due to a hydrophobic interaction between the pigment and the pigment-coated cells. The electrophoretic mobility decreased gradually with an increasing pH and/or ionic strength with both addition and no addition of the pigment derivatives. The pattern of change of the pigment adsorption ratio under varying pH and/or ionic strength values was similar to the pattern for electrophoretic mobility.     

The performance evaluation of Eugenol and Linalool microencapsulated by PLA on their activities against pathogenic bacteria

The essential oils (EOs) have great potential as a natural alternative to preserve foods against spoilage and poisoning pathogens, and they are healthy and safe. Their incorporation in polymers has been of great interest in active packaging for preserving fresh food. This work aims to evaluate the effect of encapsulation of two different oils (linalool and eugenol) as antimicrobial agent activity against Escherichia coli, Salmonella enterica subsp. enterica serovar Choleraesuis, Staphylococcus aureus, and Listeria monocytogenes. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) evaluated the EOs and their capsules with polylactic acid biopolymer. It was analyzed using the agar disc-diffusion testing method to determine the inhibition zones. The active release curves were constructed to elucidate activity efficiency. The minimum bactericidal concentration (MBC) values on E. coli, Salmonella, S. aureus, and L. monocytogenes were 0.39%, 3.13%, 0.78%, 1.56%, and 0.39%, 12.50%, 0.39%, and 12.50%, for the eugenol and linalool, respectively. To eugenol against E. coli (60 mm) and linalool against Salmonella (32 mm) exhibited relevant inhibition zones results. The EOs released promoted the inhibition zone by the volume of the EOs released over 24 h (2.2 μL/L to eugenol and 1.5 μL/L to linalool). Moreover, UV–vis results determined the active release reporting that the capsules have a prolonged efficiency, continuing to release the active up to 40 days, indicating potential application in active food packaging, extending the shelf life of food.     

Palladium-platinum electrocatalysts for the ethanol oxidation reaction: comparison of electrochemical activities in acid and alkaline media

Journal of Solid State Electrochemistry - Carbon-supported Pd, Pt, Pt1Pd1, and Pt3Pd1 electrocatalysts were prepared by metal ion chemical reduction with borohydride. The electrocatalysts were...     

A comparison of the capacity of FA-ICP-MS and FA-INAA

The platinum-group elements (PGEs) are commonly determined by INAA and ICP-MS after a NiS fire assay preconcentration. The results of the initial round robin for the PGEs and gold were examined for geological Canadian reference materials (WGB-1, TDB-1, UMT-1, WPR-1, WMG-1, and WMS-1). The Au accuracy is generally within 15% for both methods. For Ir, Os, Pd, Pt and Rh the accuracy for most samples is better than 10% for FA-ICP-MS and FA-INAA (true only for sulfide-bearing samples in the case of FA-INAA). Ru is not very accurate by either methods. Ru and Au have problems with precision which is interpreted to be related to the loss of gold in the dissolution step and for Ru, the source of the problem is not yet understood. Kurtosis show that FA-INAA has higher clustering than FA-ICP-MS for most analytes. It suggests a slightly better precision for FA-INAA. This is explained by the robustness of INAA after the NiS preconcentration despite its lower instrumental precision versus the complex dissolution steps involved in ICP-MS. For samples richer in PGEs (sulfide- and/or oxide-bearing rocks) both methods perform adequately but for low PGEs concentration samples (crustal rocks) ICP-MS shows an advantage.     

A new approach to construct full-length glycosylphosphatidylinositols of parasitic protozoa and [4-deoxy-Man-III]-GPI analogues

A new [2 + 2 + 2] approach to construct GPI molecules through the efficient synthesis of glucosamine-inositol and tetramannose intermediates led to a total synthesis of a GPI-anchor of Trypanosoma cruzi, and also afforded a key intermediate for the synthesis of valuable [4-deoxy-Man-III]-GPI analogues.     

几种磺酸铜盐的合成、表征及其催化氯乙酸酯化反应性能比较

酯化反应广泛应用于有机、生物有机以及相关的精细化学品的合成。在各类催化剂中,Lewis酸兼有均相和多相催化剂的优点。但是部分Lewis酸仍存在一些不足,如铪盐和锆盐虽然具有较高的催化活性,     

A comparison of the performance of calorimeters

Isothermal calorimetry is finding extensive application in a number of research areas. This popularity is reflected in the number of commercially available instruments which are capable of yielding a variety of thermodynamic and kinetic parameters. Whilst there has been much discussion of ways in which to validate any values returned from these instruments very little has been done quantitatively to compare the relative performances of different instruments. This paper highlights the use of a test and reference reaction quantitatively to compare the performance of three instruments (Thermometric TAM, THT μRC and a Setaram HSDSC III); the specifications of these instruments provide a range from high-sensitivity, long equilibration time to lower-sensitivity, short equilibration time. The comparison is made through a statistical analysis of values returned for the rate constant, enthalpy of reaction and activation energy for the base catalysed hydrolysis of methyl paraben. The statistical analysis from the data set discussed here indicates that there is no significant difference between the returned thermodynamic and kinetic parameters from the TAM and μRC. The analysis revealed however that the HSDSC returns values for the rate constant which are significantly different from both the TAM and μRC, although it is noted that this instrument was not specifically designed to operate in a step-isothermal mode and that it was possible to apply a correction to the data. In all cases the enthalpy data returned from all instruments were statistically similar although the μRC and HSDSC returned values which were, for the rate constant and activation energy, less precise than those obtained from the TAM. As well as highlighting the importance of using test and reference reactions, this study also shows that proper instrument selection is an important factor when designing a calorimetric experimental series.     

A comparison of Lisp and Prolog

The computer languages Prolog and Lisp are described, followed by an evaluation and comparison.     

A comparison of the photoelectron spectra of the trimethyl- and triallyl-orthoformates

The Hel photoelectron spectra of trimethyl-orthoformate and triallyl-orthoformate show low ionization potential bands arising from oxygen lone-pair n electrons and ethylenic π-bond electrons. Analysis of the spectra assisted by the use of SPINDO calculations and a density-of-states analysis indicates that structural features of the two molecules can be identified with particular regions of each spectrum. Small n-n and π-π interactions are observed, but n-π interactions are probably negligible. A conformation of TGG is indicated for the trimethyl-orthoformate in disagreement with the conclusion of an infrared spectroscopic study.