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Inhibition of blueberry PPO activity by sodium benzoate, potassium sorbate and potassium metabisulfite and their influence on degradation of individual anthocyanins in an extract was studied. Maceration of blueberries was carried out at 55ºC for 1h with the addition of 0.1% sodium benzoate or with blanching pretreatment at 90ºC for 1min. After maceration pretreatments the extracted juice was processed with traditional hot fill pasteurization, high hydrostatic pressure (HHP) and pulsed electric field (PEF). Sodium benzoate and potassium metabisulfite were very effective PPO inhibitors in concentrations of 0.1% and 10ppm, respectively. Potassium sorbate was the weakest inhibitor, with 50% PPO remaining. Degradation of anthocyanins by PPO was dependent on their structure. Tri-phenolic anthocyanins experienced the most degradation, followed by diphenolic and monophenolic compounds, respectively. Sodium benzoate was the most effective at preventing anthocyanin degradation; potassium metabisulfite did not have any protective effect, while potassium sorbate increased anthocyanin degradation Blanching of blueberries inactivated native PPO, but also increased the degradation of anthocyanins, especially malvidin glycosides. Addition of 0.1% sodium benzoate decreased PPO activity when compared to frozen blueberries but not in respect to control maceration. Only 12% of anthocyanins and 33-41% of phenolics were extracted into juice from the frozen fruit. Hot fill pasteurization, high hydrostatic pressure and pulsed electric field did not significantly influence anthocyanins, phenolics and antioxidant activity in blueberry juice.

Ce travail concerne l'extraction d'enzymes de la famille des polyphénol oxydases à partir de champignons, leur caractérisation biochimique et leur immobilisation dans des matrices solides. Ces enzymes ont tout d'abord été extraites du champignon de Paris (Agaricus bisporus) puis partiellement purifiées. Une étude de leur activité enzymatique, de leur domaine de stabilité et de leur comportement thermique a été effectuée, ainsi que l'identification d'inhibiteurs. Cette approche a été étendue à la polyphénol oxydase de la truffe de désert (Terfezia leonis Tul.). Ces deux enzymes ont ensuite été piégées dans des gels de silice pour le dosage de la dopamine par un biocapteur optique et dans un gel d'alginate pour la dégradation du phénol. / This work is devoted to the extraction of enzymes belonging to the polyphenol oxidase family from mushrooms, their biochemical characterization and their immobilization in solid hosts. These enzymes were first extracted from Paris mushrooms (Agaricus bisporus) and partially purified. A study of their enzymatic activity, stability conditions and thermal behavior was performed, together with the identification of inhibitors. A similar approach was applied to polyphenol oxidase extracted from desert truffle (Terfezia leonis Tul.). These enzymes were then trapped in silica gels for dopamine determination using an optical biosensor and in an alginate gel for phenol degradation.

Copper metallothionein (Cu-MT) was extracted from the induced biomass of Aspergillus niger. The crude extract (FI), obtained by cell homogenization, was partially purified by heat treatment (FII) and ultrafiltration (FIII). Further purification of the Cu-MT extract by affinity chromatography resulted in three major fractions, FIVa, FIVb and FIVc, of which fraction FIVc was considered to be the Cu-MT extract fraction. Fraction FIVc was re-chromatography on affinity chromatography and the eluted fraction showed a single peak (FIVc'). Spectrophotometric analysis of fraction FIVc' demonstrated a maximum absorption peak at 268 nm. Native and denatured electrophoretic analysis of fraction FIVc ' showed the presence of a single band with an estimated molecular weight of 9.5 and 10.0 kDa, respectively. Inhibition of mushroom tyrosinase (PPO) by the Cu-MT extracts was investigated, using selected phenolic substrates, including catechin, chlorogenic acid, catechol, 4-methylcatechol, caffeic acid, L-3,4-dihydroxyphenylalanine, 3,4-dihydroxyphenylacetic acid, 3-( p-hydroxyphenyl) propionic acid, p-hydroxyphenylpyruvic acid, p- and m-cresol. The results showed that the inhibitory effect of the Cu-MT extract increased with the degree of purification. The results revealed that the Cu-MT extracts were effective inhibitors of PPO activity and the best inhibitory effect was demonstrated with catechin as substrate; however, PPO activity was not inhibited by the Cu-MT extract when p-hydroxyphenylpyruvic acid and p- and m-cresol were used as substrates. The results also showed that the Cu-MT extracts exhibited different types of inhibition, including mixed, competitive and uncompetitive on PPO activity. In addition, the experimental findings indicated that the nature and degree of enzymatic inhibitions by the Cu-MT extracts were dependent upon the structural nature of the substrates as well as the methods including, spectrophotometer and polarograph, used for the detection of enzyme

Two natural enzymatic browning inhibitors, copper-metallothionein (Cu-MT) and polyphenol esterase (PPE), were obtained from A. niger and investigated. Reflectance measurements, expressed as L (lightness variable) and a (red to green degree of color) were used to compare, over extended periods of time, the relative inhibitory effectiveness of Cu-MT and PPE to those observed with the use of selected chemicals including ascorbic acid (AA), citric acid (CA), ethylenediaminetetraacetic acid (EDTA), sodium bisulfite (NaHSO3) and 4-hexylresorcinol (4HR), in the prevention of browning on the cut surfaces of selected food products such as apple and potato slices as well as freshly prepared apple juice. Treatment of each food product required an optimum concentration of the selected inhibitor for the inhibition of browning. (Abstract shortened by UMI.)

A crude enzyme extract (FI) of polyphenol esterase (PPE), obtained from the microbial culture of Aspergillus niger, was partially purified by ammonium sulfate precipitation. The partially purified fraction (FII) was subjected to further purification by ion-exchange chromatography, which resulted in five separated fractions, FIIIa, FIIIb, FIIIc, FIIId and FIIIe), where FIIIa showed the highest PPE activity towards chlorogenic acid, as substrate. The biocatalysis of the PPE with a wide range of mono- and diphenols, as substrates, was shown to inhibit mushroom tyrosinase (PPO) activity. Fraction FIIIa exhibited an inhibitory effect, measured spectrophotometrically, on PPO activity with the monophenols, including 4-hydroxyphenylpyruvic acid and m- and p-cresols and the diphenols, including chlorogenic acid, catechin, 3,4-dihydroxyphenylacetic acid (DHPAA), L-3,4-dihydroxyphenylalanine (L-DOPA), 4-methylcatechol, catechol and caffeic acid; however, using the polarographic method, the inhibition of PPO activity by PPE biocatalysis occurred with the diphenols but not with the monophenols. The selected enzymatic fraction FIIIa was further purified, using size-exclusion chromatography, which resulted in three fractions FIVa, FIVb and FIVc. Although fraction FIVc contained the highest PPE activity, it showed a lack of enzyme stability. Fraction FIIIa was therefore, subjected to further purification by hydrophobic interaction chromatography thereby yielding fractions FVa, FVb, FVc, FVd, FVe, FVf and FVg, where fraction FVc showed the highest PPE activity. The denatured electrophoretic analysis of fraction FVc showed the presence of one major band, with a molecular weight of 60 kDa. The successive purification of PPE resulted in a marked increase in the inactivation of PPO activity with diphenols, as demonstrated by both the lower I50 and inhibition dissociation constant (Ki) values. The purified fraction FVc was shown to exhibit, spectrophotometrically, a competitive and un

The latest generation of fighter aircraft utilizes a 270Vdc power system [1]. Such high voltage DC power systems are difficult to protect with conventional circuit breakers because the current does not automatically go to zero twice per cycle during a fault like it does in an AC power system and thus arcing of the contacts is a problem. Solid state power controllers (SSPCs) are the solid state equivalent of a circuit breaker that do not arc and which can respond more rapidly to a fault than a mechanical breaker [2]. Present SSPCs are limited to lower voltages and currents by the available power semiconductors [8,9]. This dissertation presents design and experimental results for a SSPC that utilizes SiC power JFETs for the SSPC power switch to extend SSPC capability to higher voltages and currents in a space that is smaller than what is practically achievable with a Si power switch. The research started with the thermal analysis of the SSPCs power switch, which will guide the development of a SiC JFET multi-chip power module to be fabricated by Solid State Devices Inc. (SSDI) using JFETs from SiCED and/or Semisouth LLC. Multiple multi-chip power modules will be paralleled to make the SSPC switch. Fabricated devices were evaluated thermally both statically and dynamically and electrically both statically and dynamically. In addition to the SiC module research a detailed design of the high voltage SSPC control circuit capable of operating at 200andamp;ordm;C was completed including detailed analysis, modeling and simulations, detailed schematic diagrams and detailed drawings. Finally breadboards of selected control circuits were fabricated and tested to verify simulation results. Methods for testing SiC JFET devices under transient thermal conditions unique to the SSPC application was also developed.

A greenhouse experiment was carried out to screen for potential effects of transgenic aspen over-expressing a hybrid poplar leaf polyphenol oxidase gene on rhizosphere communities. Heterotrophic plate counts and cultivation-independent methods were used to compare bacterial and fungal populations associated with transgenic PPO over-expressing and unmodified control trees. Total community DNA extracted from rhizosphere soils was used to establish Iibraries containing partial gene sequences that were PCR-amplified from community members, and putative taxonomy was assigned to clones based on similarity to reference sequences. Gene libraries for the bacterial component of the rhizosphere were established using partial 16S rRNA and chaperonin-60 gene sequences, and the fungal community was characterized based on partial 18S rRNA gene sequences. Phylogenetic analysis revealed that bacterial 16S gene libraries were dominated by Alphaproteobacterial sequences, and the CPN-60 gene libraries were dominated by members of the Bacteroidetes/Chlorobi group, illustrating the biases potentially incurred by using a single gene locus to profile microbial diversity. In both CPN-60 and 16S rRNA libraries, only minor components of the bacterial community differed between transgenic and unmodified trees. Comparisons based on library coverage indicated that changes in bacterial community structure between transgenic and unmodified trees were minor in comparison to differences observed between individual trees of the same type, and no significant differences in terms of bacterial species diversity were revealed by the calculated diversity, dominance and evenness indices. In comparison to the bacterial gene libraries, higher coverage of the underlying population was achieved in the fungal 18S libraries. Members of the Zygomycota, Chytridiomycota, Ascomycota, and Basidiomycota were recovered from both libraries. Dominant groups of fungi associated with each tree type were highly similar, although there were some qualitative differences in the recovery of less abundant fungi as a result of the underlying heterogeneity of the fungal population. No clear differences in terms of fungal species richness were associated with transgenic or unmodified trees, although control libraries were characterized by a slightly higher level of dominance. In general, the methods employed revealed only minor differences between the bacterial and fungal communities associated with transgenic and unmodified trees, suggesting that impacts of the transgenic plants on the rhizosphere community were minimal.

Digestive proteases were isolated from the pancreas of the stomachless cunner fish (Tautogolabrus adspersus) and characterized in terms of their physicochemical properties, their ability to hydrolyze native pectin methylesterase (PME) from orange and polyphenol oxidases (PPO) from mushroom and the ability of the cunner enzyme(s) to maintain the stability of orange juice cloud. / The cunner trypsin fraction exhibited exceptional capacity to hydrolyze native proteins versus the bovine trypsin. Incubation of native PME with cunner or bovine trypsin resulted in a loss of 75% or 35% in PME activity respectively. Similarly, a 75% or 55% loss in PPO activity was observed after treatment with cunner and bovine trypsin respectively. Bovine trypsin, however, hydrolyzed the heat-denatured PME and PPO better than the cunner trypsin. Also, there was no reactivation of both PME and PPO activity after treatment with either the cunner or bovine enzyme during storage at 4$ sp circ$C for 3 weeks. However, PPO retained up to 20% or 50% of the initial activity after treatment with cunner or bovine trypsin, respectively. / A 3 x 3 factorial design involving the factors of temperature, enzyme concentration and incubation time carried out gave an r$ sp2$ of 0.92 and 0.95 for cunner and bovine trypsin treated PME respectively. On the other hand, an r$ sp2$ of 0.91 and 0.94 was obtained for the combined effects using cunner and bovine trypsin for PPO inactivation. Validation of the model of PME inactivation measured as the % cloud remaining revealed that the cunner trypsin fraction upheld the cloud stability of cloud juice better than bovine trypsin, with cunner trypsin retaining more than 90% of the cloud whereas the juice treated with bovine trypsin only resulted in a 70% retention of the juice cloud. (Abstract shortened by UMI.)

No description available.