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PubMed2003

Application of partial least-squares regression to near-infrared reflectance spectroscopic determination of shive content in flax.

Shive, the nonfiberous core portion of the stem, in flax fiber after retting is related to fiber quality. The objective of this study is to develop a standard calibration model for determining shive content in retted flax by using near-infrared reflectance spectroscopy. Calibration samples were prepared by manually mixing pure, ground shive and pure, ground fiber from flax retted by three different methods (water, dew, and enzyme retting) to provide a wide range of shive content from 0 to 100%. Partial least-squares (PLS) regression was used to generate a calibration model, and spectral data were processed using various pretreatments such as a multiplicative scatter correction (MSC), normalization, derivatives, and Martens' Uncertainty option to improve the calibration model. The calibration model developed with a single sample set resulted in a standard error of 1.8% with one factor. The best algorithm was produced from first-derivative processing of the spectral data. MSC was not effective processing for this model. However, a big bias was observed when independent sample sets were applied to this calibration model to predict shive content in flax fiber. The calibration model developed using a combination sample set showed a slightly higher standard error and number of factors compared to the model for a single sample set, but this model was sufficiently accurate to apply to each sample set. The best algorithm for the combination sample set was generated from second derivatives followed by MSC processing of spectral data and from Martens' Uncertainty option; it resulted in a standard error of 2.3% with 2 factors. The value of the digital second derivative centered at 1674 nm for these spectral data was highly correlated to shive content of flax and could form the basis for a simple, low-cost sensor for the shive or fiber content in retted flax.

Algorithms↗
PubMed2004

Influence of carbon and nitrogen sources and temperature on hyperproduction of a thermotolerant beta-glucosidase from synthetic medium by Kluyveromyces marxianus.

The effect of carbon source and its concentration, inoculum size, yeast extract concentration, nitrogen source, pH of the fermentation medium, and fermentation temperature on beta-glucosidase production by Kluyveromyces marxianus in shake-flask culture was investigated. These were the independent variables that directly regulated the specific growth and beta-glucosidase production rate. The highest product yield, specific product yield, and productivity of beta-glucosidase occurred in the medium (pH 5.5) inoculated with 10% (v/v) inoculum of the culture. Cellobiose (20 g/L) significantly improved beta-glucosidase production measured as product yield (YP/S) and volumetric productivity (QP) followed by sucrose, lactose, and xylose. The highest levels of productivity (144 IU/[L.h]) of beta-glucosidase occurred on cellobiose in the presence of CSL at 35 degrees C and are significantly higher than the values reported by other researchers on almost all other organisms. The thermodynamics and kinetics of beta-glucosidase production and its deactivation are also reported. The enzyme was substantially stable at 60 degrees C and may find application in some industrial processes.

Animals↗
PubMed2002

Increasing fermentation efficiency at high sugar concentrations by supplementing an additional source of nitrogen during the exponential phase of the tequila fermentation process.

In the tequila industry, fermentation is traditionally achieved at sugar concentrations ranging from 50 to 100 g x L(-1). In this work, the behaviour of the Saccharomyces cerevisiae yeast (isolated from the juices of the Agave tequilana Weber blue variety) during the agave juice fermentation is compared at different sugar concentrations to determine if it is feasible for the industry to run fermentation at higher sugar concentrations. Fermentation efficiency is shown to be higher (above 90%) at a high concentration of initial sugar (170 g x L(-1)) when an additional source of nitrogen (a mixture of amino acids and ammonium sulphate, different than a grape must nitrogen composition) is added during the exponential growth phase.

Alcoholic Beverages↗
PubMed2005

Nickel recovery from spent nickel catalyst.

A process for nickel recovery from a spent catalyst of definite composition has been developed using the hydro-metallurgical route. The processing steps includes direct sulphuric acid leaching followed by separation of iron as well as silica and other impurities. For a 152 microm particle size catalyst, extraction of about 98% nickel was achieved at 363 K in 2 h using a sulphuric acid concentration (v/v) of 8% and a pulp density of 10%. The dissolution of nickel followed diffusion-controlled leaching kinetics. Increase in temperature and sulphuric acid concentration resulted in increase in the nickel recovery. The activation energy for nickel dissolution was calculated to be 62.8 kJ mol(-1). Finally, nickel was recovered as value-added products such as sulphide and oxalate with overall recovery of 90 and 88% of nickel, respectively.

Catalysis↗
PubMed2002

[Recovery process of nitric acid, copper and nickel in deplating wastewater].

The recovery process of nitric acid, copper and nickel in deplating wastewater was developed by using the combined technique of distillation, solvent extraction and precipitation. The conditions of the separation of copper and nickel by solvent extraction using P507 in kerosene and stripping copper with H2SO4 were specially investigated and the optimal parameters were determined. The results of experiment indicated that the recovery ratio of nitric acid was 97.8%, and under the optimized conditions of extraction process, concentration of original effluence ranged in 15-20 mg/mL copper, 5-10 mg/mL nickel, pH 1-2, concentration of extractant was 35% (V/V), saponification degree was 60%, phase ratio was 1:1, reaction time was 2 min, temperature ranged in 20 degrees C-25 degrees C, the one stage extraction efficiency of copper was higher than 90%, the separation ratio of copper and nickel was up to 75; copper and nickel could be completely separated by a continuous countercurrent three-stage extraction. The nickel could be recovered from the water phase by precipitating with NaOH and the recovery ratio of nickel reached up to 99.9% by controlling pH in solution within 10-11. After these treatment, the effluent could meet the national standards of wastewater discharge.

Copper↗
PubMed2004

Identification of surface states on finely divided supported palladium catalysts by means of inelastic incoherent neutron scattering.

The purpose of the present investigation was to utilize the inelastic incoherent neutron scattering (INS) technique to reveal changes at the surface of technical catalysts under the influence of hydrogen in gas/solid interactions and during chemical reactions in a liquid-phase process. The formation and the properties of supported palladium hydride and changes of the hydrogen-related surface chemistry of the corresponding activated carbon supports in 20% Pd/C catalysts after short-term and long-term hydrogen cycling at different hydrogen pressures and temperatures were studied. The spectra indicate that hydrogenation of the activated carbon support by hydrogen spillover occurs to, partly, give a material that strongly resembles a-C:H (amorphous hydrogenated carbon). Indications for different relaxation phenomena and long-range phase coherence inside of supported particles of palladium hydride compared to hydrogenated palladium black were obtained. A 5% Pd/C catalyst after use in C-C coupling reactions, the Heck reaction of bromobenzene and styrene to stilbenes, was also studied after subsequent solvent extraction. Evidence for a preferential adsorption and accumulation of cis-stilbene at the catalyst surface was obtained. INS allows identification of a certain isomer from a complex reaction mixture preferentially adsorbed at the surface of a finely divided industrial heterogeneous catalyst.

Adsorption↗
PubMed2006

A multiplex real-time PCR assay for detection of Xanthomonas campestris from brassicas.

AIMS: To develop a sensitive real-time PCR-based protocol for the detection of Xanthomonas campestris pathovars from Brassica seed. METHODS AND RESULTS: A 5' nuclease real-time PCR assay was developed to screen Brassica spp. seed for the presence of X. campestris pathovars that cause black rot. The assay amplifies a 78-bp segment of the X. campestris hrpF gene and a 100-bp segment of the Brassica spp. 18S-25S internal transcribed spacer region. The Brassica spp. target provides an internal control for the amplification process to prevent false negatives that may arise from inhibitors that are often present in extracts from plant material. Whilst the primers were compatible with SYBR Green I assays, the use of fluorescently labelled probes in a 5' nuclease assay afforded greatest sensitivity and specificity. Seed batches carrying one artificially infected seed among 10,000 were readily detected using the assay. The multiplex real-time PCR assay permitted the rapid detection of pathogenic strains of X. campestris from bacterial colonies, Brassica seed and plants. CONCLUSIONS: Strains of X. campestris pathogenic to brassicas were readily detected from seed via a multiplex 5' nuclease real-time PCR assay. The real-time assay offers an improvement in sensitivity and a reduced turn-around time over the conventional multiplex PCR. SIGNIFICANCE AND IMPACT OF THE STUDY: Real-time PCR can be used to rapidly screen Brassica spp. seed batches for the presence of X. campestris pathovars. This assay provides a means for growers and the seed industry to be aware of the black rot status of their planting material, so that they may more effectively employ disease control measures or seed disinfection.

Brassica↗
PubMed2004

[Antioxidative activity of propolis from Dalmatia (Croatia)].

AIM: The aim of this study was to determine the antioxidative activity of propolis from ecologically clean parts of Dalmatia. METHODS: Phenol concentration in ethanolic propolis extracts was determined by Folin-Ciocalteu reagent using gallic acid as the standard. Flavonoid phenolic compounds were analyzed after precipitation with formaldehyde. The residual non-flavonoid phenolics were also determined by Folin-Ciocalteu method. By determining the change of peroxide number (PN), of tiobarbiture acid reactive species (TBARS), and of DPPH-radical activity, antioxidative efficiency of propolis was tested and compared with well known and widely used synthetic antioxidants. Values of PN and TBARS were determined at 60 degrees C in samples of trigyceride substrate (lard) without and with the addition of antioxidants. Compared was the efficiency of three antioxidants: propolis (alcoholic extract), vitamin E, and (+)-catechin in a concentration of 1%. PN was monitored during 50 days. By the method of Sedlacek, TBARS were measured during 30 days. Antioxidative activity of propolis extract was also measured in terms of hydrogen donating ability using stable radical alpha,alpha-diphenyl-beta-picril hidrazyl (DPPH*) and compared with commercial synthetic antioxidants of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and (+)-cathecin. Inhibition degree of DPPH* was calculated by the formula of Yen and Duh. RESULTS: Total phenol content, expressed as gallic acid, in propolis extracts varied from 75.2 to 90.2 g/kg propolis. The proportion of flavonoids in total phenols ranged from 62% to 65%. Values of TBARS were not increased only in samples with added propolis. The inhibition of DPPH-radical by propolis extracts ranged from 93% to 96%, by catechin 95%, by BHT 49%, and by BHA 64%. Compared to BHT and BHA, propolis extracts showed greater reducing activity against DPPH-radical. DISCUSSION: The chemical composition of propolis, and thus its biological activity depend on the plant from which it has been collected, and on the macro- and microclimatic conditions. Many compounds in propolis exert antioxidative activity. A belief was expressed that the biological activity of propolis is very probably based mostly on its antioxidative efficiency. Dalmatian propolis showed high efficiency in the prevention of oxidative processes. This could be explained by the high proportion of polyphenol constituents, especially flavonoids. A very low and equal degree of increase of PN, as a measure of oxidative processes, was noticed in the samples of triglyceride substrate with the addition of propolis and (+)-catechin. The greatest rise of TBARS was measured in the samples of pure lard. There was no increase of TBARS only in the samples with added propolis. Propolis and (+)-catechin showed great efficiency in the inhibition of DPPH-radical, greater than BHT and BHA, which are widely used in food industry. CONCLUSION: The results indicate that Dalmatian propolis could be an efficient protective agent against oxidative processes in food. The high antioxidative activity of propolis, its natural origin, and present knowledge about its biological properties, make it a very promising nutritional additive for human diet.

Antioxidants↗
PubMed2000

Changes of soil and plant tissue selenium status in an upland grassland contaminated by selenium-rich agricultural drainage sediment after ten years transformed from a wetland habitat.

A field survey was conducted in 1989, 1994, and 1999 in order to monitor the soil and vegetation Se concentrations at the Kesterson upland grassland contaminated by Se-rich drainage sediment. The rate of Se dissipation estimated by the change of soil Se concentration, via volatilization, found to be about 1.1% per year. Soil water-extractable Se increased in 1994, but greatly reduced in 1999. The increase of soil Se concentration in the top 15 cm of soil at the fresh-soil fill sites indicates that the plants were able to effectively take up the soluble soil Se from the lower soil profile and to deposit it on the top of the field. This process may reduce the rate of leaching of soil Se. Overall, the concentration of soil water-soluble Se was relatively low and it is unlikely that problems of transport of Se from the Kesterson soil to the adjacent uncontaminated environment by leaching can occur. Plant tissue Se concentration was found to coincide with the soil water-extractable Se concentration. The average plant tissue Se concentration and soil water-extractable Se detected in 1999 was about 10 microg Se g(-1) and 110 microg Se kg(-1) dry weight, respectively, and the estimated bioaccumulation value of this upland grassland is less than 10% of the previous wetland habitat. Therefore, the existing Kesterson grassland should not be at high risk to the environment.

Agriculture↗
PubMedDate not supplied

Gas chromatographic determination of fenpropimorph residues in citrus fruit.

A rapid gas chromatographic method for determining fenpropimorph residues in citrus fruit is reported. The fungicide is extracted with hexane after pH adjustment of the fruit homogenate. A short liquid-liquid partitioning process is performed before gas chromatography on an OV-17 column with nitrogen-phosphorus specific detection. The limit of detection of the method was 0.01 mg/kg, based on a 25 g sample. Recovery was always higher than 70%. Fenpropimorph residues in "Washington Navel" oranges and "Hernandina" clementine fruits dipped in a 1500 mg/L fungicide solution were determined. The fungicide remains mainly in the peel, with levels less than 0.1 mg/kg in the pulp. Fungicide residues in the peel decrease during storage, mainly in Washington Navel peel, where values decreased from 5.2 to 2.8 mg/kg.

Chromatography, Gas↗
PubMed2003

Microwave-assisted steam distillation with simultaneous liquid/liquid extraction of pentachlorophenol from organic wastes and soils.

An efficient method for extracting pentachlorophenol (PCP) from organic solid matrices and soils using direct excitation of the fresh samples by microwave energy is presented. Steam-volatile PCP partitioned into the organic solvent n-hexane within the closed extraction vessel, resulting in recovery rates of 85-92% from aged samples. Condensed water provided a boundary layer between extract and sample thus preventing any contact of PCP with and re-partitioning into the sample. Duration of the microwave-assisted process was 35 min. Crude extracts were derivatized by heptafluorobutyric anhydride (HFBA) to improve selectivity and sensitivity resulting in detection limits of 1-2 micro g kg(-1). GC-MS/MS analyses proved that suitable extracts with only minor impurities were obtained. Common sample treatment steps like drying, thorough grinding, frequent transferring, and tedious clean-up, and concentration procedures which all can cause certain losses of analyte were minimized. The efficiency of the method was verified by comparison with an established ultrasonic extraction procedure. This microwave-assisted pressurized steam distillation with simultaneous partition into an organic phase thus provides a streamlined and efficient strategy which requires no additional investment in standard equipment for microwave-assisted extractions (MAE). Degradation of analytes at longer extraction times must be taken into account.

Chemical Fractionation↗
PubMed2003

Commercialization of a novel fermentation concept.

Fermentation is the core of biotechnology where current methodologies span across technologies based on the use of either solid or liquid substrates. Traditionally, solid substrate fermentation technologies have been the widely practiced in the Far East to manufacture fermented foods such as soya sauce, sake etc. The Western World briefly used solid substrate fermentation for the manufacture of antibiotics and enzymes but rapidly replaced this technology with submerged fermentation which proved to be a superior technology in terms of automation, containment and large volume fermentation. Biocon India developed its enzyme technology based on solid substrate fermentation as a low-cost, low-energy option for the production of specialty enzymes. However, the limitations of applying solid substrate fermentation to more sophisticated biotechnology products as well as large volume fermentations were recognized by Biocon India as early as 1990 and the company embarked on a 8 year research and development program to develop a novel bioreactor capable of conducting solid substrate fermentation with comparable levels of automation and containment as those practiced by submerged fermentation. In addition, the novel technology enabled fed-batch fermentation, in situ extraction and other enabling features that will be discussed in this article. The novel bioreactor was christened the "PlaFractor" (pronounced play-fractor). The next level of research on this novel technology is now focused on addressing large volume fermentation. This article traces the evolution of Biocon India's original solid substrate fermentation to the PlaFractor technology and provides details of the scale-up and commercialization processes that were involved therein. What is also apparent in the article is Biocon India's commercially focused research programs and the perceived need to be globally competitive through low costs of innovation that address, at all times, processes and technologies that exhibit high degrees of conformance to the international standards of regulatory and good manufacturing practice.

Bioreactors↗
PubMed2001

Evaluation of parallel operated small-scale bubble columns for microbial process development using Staphylococcus carnosus.

Shake flasks and pH-controlled small-scale bubble columns were compared with respect to their usefulness as a basic tool for process development for human calcitonin precursor fusion-protein production with Staphylococcus carnosus. Parallel control of the pH (and making use of the base addition data) is necessary to study the effects of medium composition, to identify pH-optima and to develop a medium, which minimizes the acid excretion of S. carnosus. This medium with glycerol as energy source and yeast extract as carbon and nitrogen source resulted in cell dry weight concentration in shake flasks of 5 g l(-1), which were thus improved by a factor of 10. Cell dry weight concentrations of up to 12.5 g l(-1) were measured in the batch process with pH-controlled small-scale bubble columns due to their higher oxygen transfer capability. In contrast to shake flasks it was demonstrated, that the batch process performance of recombinant S. carnosus secreting the human calcitonin precursor fusion-protein was identical within the estimation error in pH-controlled small-scale bubble columns compared to the stirred-tank reactor.

Cell Division↗
PubMedDate not supplied

Fusion tails for the recovery and purification of recombinant proteins.

Several fusion tail systems have been developed to promote efficient recovery and purification of recombinant proteins from crude cell extracts or culture media. In these systems, a target protein is genetically engineered to contain a C- or N-terminal polypeptide tail, which provides the biochemical basis for specificity in recovery and purification. Tails with a variety of characteristics have been used: (1) entire enzymes with affinity for immobilized substrates or inhibitors; (2) peptide-binding proteins with affinity to immunoglobulin G or albumin; (3) carbohydrate-binding proteins or domains; (4) a biotin-binding domain for in vivo biotination promoting affinity of the fusion protein to avidin or streptavidin; (5) antigenic epitopes with affinity to immobilized monoclonal antibodies; (6) charged amino acids for use in charge-based recovery methods; (7) poly(His) residues for recovery by immobilized metal affinity chromatography; and (8) other poly(amino acid)s, with binding specificities based on properties of the amino acid side chain. Fusion tails are useful at the lab scale and have potential for enhancing recovery using economical recovery methods that are easily scaled up for industrial downstream processing. Fusion tails can be used to promote secretion of target proteins and can also provide useful assay tags based on enzymatic activity or antibody binding. Many fusion tails do not interfere with the biological activity of the target protein and in some cases have been shown to stabilize it. Nevertheless, for the purification of authentic proteins a site for specific cleavage is often included, allowing removal of the tail after recovery.

Amino Acids↗
PubMed2004

Optimization of extracellular endoxylanase, endoglucanase and peroxidase production by Streptomyces sp. F2621 isolated in Turkey.

AIMS: To determine the effect of environmental conditions on the production of extracellular lignocellulose-degrading enzymes by Streptomyces sp. F2621 and to assess the potential use of these enzymes in the hydrolysis of lignocellulose material. METHODS AND RESULTS: The production of extracellular lignocellulose-degrading enzymes, endoxylanase, endoglucanase and peroxidase during the growth of Streptomyces sp. F2621 in basal salts-yeast extract medium containing different carbon sources and the effect of a number of environmental parameters (e.g. carbon sources and concentrations, pH and temperature) were investigated. The highest endoxylanase (22.41 U ml(-1)) and peroxidase (0.58 U ml(-1)) activities were obtained after 2-4 days of incubation at 30 degrees C in a basal salts medium containing 0.4% (w/v) oat spelt xylan and 0.6% (w/v) yeast extract, corresponding to C : N ratio of 6 : 1. Cell-free extracellular enzyme preparations from the strain were capable of releasing both sugar and aromatic compounds during incubation with eucalyptus paper pulp, straw and xylan. Overall, 9.3% hydrolysis of xylan occurred after 24-h incubation. However the rates of hydrolysis of paper pulp and straw were approximately twofold less than xylan hydrolysis, although the total percentage hydrolysis of available substrate (24.5% and 16.3%, respectively) was greater than xylan hydrolysis. CONCLUSIONS: The high levels of enzyme production achieved under batch cultivation conditions, coupled with no significant production of endoglucanase during the growth phase of organism and the release of both sugar and aromatic compounds from paper pulp and straw signify the suitability for these enzymes for industrial applications such as pulp and paper production. SIGNIFICANCE AND IMPACT OF THE STUDY: The results highlight the environmental conditions for the production of extracellular lignocellulose-degrading enzymes by Streptomyces sp. F2621 and suggest the use of streptomycetes and/or their enzymes in industrial processes.

Carbohydrate Metabolism↗
PubMed2005

Hydrolytic enzyme activities of extracted humic substances during the vermicomposting of a lignocellulosic olive waste.

Humic substances and three hydrolytic enzymes (beta-glucosidase, phosphatase and urease) were extracted by neutral sodium pyrophosphate from an olive waste (dry olive cake), alone or mixed with municipal biosolids, during a nine month vermicomposting process. Easily degradable compounds decreased during the vermicomposting process because of microbial consumption. When municipal biosolids were added to dry olive cake, microbial activity increased and the amounts of compounds extracted by pyrophosphate were three times lower than olive cake alone. In both instances, beta-glucosidase, phosphatase and urease activities of the organic extracts either increased or remained the same after a nine month period of vermicomposting, thus suggesting that the humus enzyme complexes resisted microbial and earthworm attack. It is known that humus immobilised enzymes also remain active in soil environments, reactivating the nutrient cycles in soil. The use as amendments of vermicomposted olive cake, alone or when mixed with biosolids, could be a good alternative to reactivate the C, P and N-cycles in degraded soils for regeneration purposes.

Cellulose↗
PubMed2004

Selective adsorption of polychlorinated dibenzo-p-dioxins and dibenzofurans by the zeosils UTD-1, SSZ-24, and ITQ-4.

Zeosils are microporous solids with a pure silica framework. Due to their hydrophobic properties, zeosils are ideal host materials for the adsorption of hydrophobic guest molecules. We tested zeosils with different pore diameters (UTD-1, SSZ-24 and ITQ-4 as well as CIT-5) for the selective adsorption of the polychlorinated dibenzo-p-dioxins and dibenzofurans. This group of highly toxic substances contains 210 congeners that possess similar chemical properties, but differ in their size and shape. In the experiment, polychlorinated dibenzo-p-dioxins and dibenzofurans were extracted from fly ash of a waste incinerator, adsorbed on amorphous silica, then thermally desorbed and flushed over a sequential arrangement of the zeosils at elevated temperature by a stream of nitrogen. ITQ-4 with the smallest pore diameter was placed first, followed by SSZ-24 and, finally, by UTD-1 with the largest pore diameter. After the experiment, the zeosils were analysed for their contents of the different congeners. The results show that the sorption of the congeners occurs selectively and that it is governed by the size and the shape of the dioxin molecules, which in turn depend on the number of chlorine atoms and the pattern of chlorine substitution (regioisomers). Geometrical reasoning as well as molecular dynamics calculations on the zeosil structures and on the dioxin molecules were helpful in rationalising the results. This work represents an especially complex case of the molecular sieving effect and may lead to a selective on-line monitoring of the concentrations of dioxin molecules in waste gases of industrial combustion processes. The size- and shape-selective sorption of dioxin molecules may also bear some resemblance to the molecular recognition process that occurs in nature at the aryl hydrocarbon receptor.

Journal Article↗
PubMed2006

Tyrosinase extract from Agaricus bisporus mushroom and its in natura tissue for specific phenol removal.

Phenols are toxic pollutants found in industrial wastes imposing several risks to human health. Tyrosinase (EC 1.14.18.1) is an oxygenase oxyreductase found in several life forms, like the mushroom Agaricus bisporus. This enzyme is readily available from this fungal tissue leading to high activity extracts without extensive purification, thus suggesting its potential as a biocatalyst for applications involving biomodification of phenols or bioremediation of phenol-polluted waters. The purpose of this work was to employ a crude extract from the Agaricus bisporus mushroom and its biomass for the removal of phenol from polluted water. Experiments were carried out without pH control. The initial phenol concentration in all solutions was 100 mg l(-1). Four enzymatic concentrations (50, 100, 200 and 400 U ml(-1)) were tested. Reactions, with 200 U ml(-1) and 400 U ml(-1) enzymatic activity, led to 90% of phenol removal. Chitosan was used as a coagulant, but no significant improvement was observed. The in natura fungi was also able to remove 90% of phenol, demostrating its viability as a biocatalyst in bioremediation process.

Agaricus↗