Production of fructooligosaccharides by the mycelia of Aspergillus japonicus immobilized in calcium alginate

This work describes fructose oligosaccharide (FOS) production by the immobilized mycelia (IM) of a strain of Aspergillus japonicus, isolated from soil. The microorganism was inoculated into 50 mi of medium composed of sugar cane molasses (5.0% of total sugars); yeast powder; 2.0%; K2HPO4, 0.5%; NaNO3, 0.2%; MgSO4. 7H(2)O, 0.05%; KCl, 0.05%, final pH 5.0, and the flasks were agitated in an orbital shaker at 200 rpm for 60 h, at 30 degrees C. The beta-fructofuranosidase activity (Uf), transfructosylating activity (Ut), hydrolyzing activity (Uh), and FOS production were analyzed by high performance liquid chromatography. FOS production was performed in a batch process in a 2-l jar fermenter by IM in calcium alginate beads. The optimum pH and temperature were 5.0-5.6 and 55 degrees C, respectively No loss of activity was observed when the mycelium was maintaned at 60 degrees C for 60 min. Maximum production was obtained using 5.75% (cellular weight/volume) of mycelia (122.4 Ut g(-1)) and 65% sucrose solution (w:v) for 4 h of reaction when the final product reached 61.28% of fetal FOS containing GF(2) (30.56%), GF(3) (26.45%), GF(4) (4.27%), sucrose (9.6%) and glucose (29.10%). In the assay conditions, 23 batches were performed without loss of activity of the IM, showing that the microorganism and the process utilized have potential for industrial applications. (C) 1998 Elsevier B.V. Ltd. All rights reserved.

Determination of pesticide residues in coconut water by liquid-liquid extraction and gas chromatography with electron-capture plus thermionic specific detection and solid-phase extraction and high-performance liquid chromatography with ultraviolet detection

Two simple methods were developed to determine, 11 pesticides in coconut water, a natural isotonic drink rich in salts, sugars and vitamins consumed by the people and athletes. The first procedure involves solid-phase extraction using Sep-Pak Vac C-18 disposable cartridges with methanol for elution. Isocratic analysis was carried out by means of high-performance liquid chromatography with ultraviolet detection at 254 nm to analyse captan, chlorothalonil, carbendazim, lufenuron and diafenthiuron. The other procedure is based on liquid-liquid extraction with hexane-dichloromethane (1:1, v/v), followed by gas chromatographic analysis with effluent splitting to electron-capture detection for determination of endosulfan, captan, tetradifon and trichlorfon and thermionic specific detection for determination of malathion, parathion-methyl and monocrotophos. The methods were validated with fortified samples at different concentration levels (0.01-12.0 mg/kg). Average recoveries ranged from 75 to 104% with relative standard deviations between 1.4 and 11.5%. Each recovery analysis was repeated at least five times. Limits of detection ranged from 0.002 to 2.0 mg/kg. The analytical procedures were applied to 15 samples and no detectable amounts of the pesticides were found in any samples under the conditions described. (C) 2002 Elsevier B.V. B.V. All rights reserved.

New crystal forms of Diocleinae lectins in the presence of different dimannosides

Studying the interactions between lectins and sugars is important in order to explain the differences observed in the biological activities presented by the highly similar proteins of the Diocleinae subtribe. Here, the crystallization and preliminary X--ray data of Canavalia gladiata lectin (CGL) and C. maritima lectin (CML) complexed with Man(alpha 1-2)Man(alpha 1)OMe, Man(alpha 1-3)Man(alpha 1)OMe and Man(alpha 1-4)Man(alpha 1)OMe in two crystal forms [the complexes with Man(alpha 1-3)Man(alpha 1)OMe and Man(alpha 1-4)Man(alpha 1)OMe crystallized in space group P3(2) and those with Man(alpha 1-2)Man(alpha 1)OMe crystallized in space group I222], which differed from those of the native proteins (P2(1)2(1)2 for CML and C222 for CGL), are reported. The crystal complexes of ConA-like lectins with Man(alpha 1-4)Man(alpha 1)OMe are reported here for the first time.

Sugarcane molasses and yeast powder used in the Fructooligosaccharides production by Aspergillus japonicus-FCL 119T and Aspergillus niger ATCC 20611

Different concentrations of sucrose (3-25% w/v) and peptone (2-5% w/v) were studied in the formulation of media during the cultivation of Aspergillus japonicus-FCL 119T and Aspergillus niger ATCC 20611. Moreover, cane molasses (3.5-17.5% w/v total sugar) and yeast powder (1.5-5% w/v) were used as alternative nutrients for both strains' cultivation. These media were formulated for analysis of cellular growth, P-Fructosyltransferase and Fructooligosaccharides (FOS) production. Transfructosylating activity (U-t) and FOS production were analyzed by HPLC. The highest enzyme production by both the strains was 3% (w/v) sucrose and 3% (w/v) peptone, or 3.5% (w/v) total sugars present in cane molasses and 1.5% (w/v) yeast powder. Cane molasses and yeast powder were as good as sucrose and peptone in the enzyme and FOS (around 60% w/w) production by studied strains.

Glass transitions for freeze-dried and air-dried tomato

Glass transition temperatures of freeze-dried tomato conditioned at various water activities at 25 C were determined by differential scanning calorimetry (DSC). Air-dried tomato with and without osmotic pre-treatment in sucrose/NaCl solutions was also analyzed. Thermograms corresponding to the low water activity domain (0.11 less than or equal to a(w) less than or equal to 0.75) revealed the existence of two glass transitions, which were attributed to separated phases formed by sugars and water and other natural macromolecules present in the vegetable. Both transitions were plasticized by water and experimental data could be well correlated by the Gordon-Taylor equation in the low-temperature domain, and by the Kwei model in the high-temperature domain. For higher water activities, the low-temperature glass transition curve exhibited a discontinuity, with suddenly increased glass transition temperatures approaching a constant value that corresponds to the T-g of the maximally freeze-concentrated amorphous matrix. The unfreezable water content was determined through the melting enthalpy dependence on the moisture content. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Structural analysis of Canavalia maritima and Canavalia gladiata lectins complexed with different dimannosides: New insights into the understanding of the structure-biological activity relationship in legume lectins

Plant lectins, especially those purified from species of the Legummosae family, represent the best studied group of carbohydrate-binding proteins. The legume lectins from Diocleinae subtribe are highly similar proteins that present significant differences in the potency/ efficacy of their biological activities. The structural studies of the interactions between lectins and sugars may clarify the origin of the distinct biological activities observed in this high similar class of proteins. In this way, this work presents a crystallographic study of the ConM and CGL (agglutinins from Canavalia maritima and Canavalia gladiata, respectively) in the following complexes: ConM/ CGL:Man(alpha 1-2)Man(alpha 1-0)Me, ConM/CGL:Man(alpha 1-O)Man(alpha 1-O)Me and ConM/CGL:Man(alpha 1-4)Man(alpha 1-O)Me, which crystallized in different conditions and space group from the native proteins.The structures were solved by molecular replacement, presenting satisfactory values for R-factor and R-factor. Comparisons between ConM, CGL and ConA (Canavalia ensiformis lectin) binding mode with the dimannosides in subject, presented different interactions patterns, which may account for a structural explanation of the distincts biological properties observed in the lectins of Diocleinae subtribe. (C) 2007 Elsevier B.V. All rights reserved.

Difference variance dispersion graphs for comparing response surface designs with applications in food technology

Variance dispersion graphs have become a popular tool in aiding the choice of a response surface design. Often differences in response from some particular point, such as the expected position of the optimum or standard operating conditions, are more important than the response itself. We describe two examples from food technology. In the first, an experiment was conducted to find the levels of three factors which optimized the yield of valuable products enzymatically synthesized from sugars and to discover how the yield changed as the levels of the factors were changed from the optimum. In the second example, an experiment was conducted on a mixing process for pastry dough to discover how three factors affected a number of properties of the pastry, with a view to using these factors to control the process. We introduce the difference variance dispersion graph (DVDG) to help in the choice of a design in these circumstances. The DVDG for blocked designs is developed and the examples are used to show how the DVDG can be used in practice. In both examples a design was chosen by using the DVDG, as well as other properties, and the experiments were conducted and produced results that were useful to the experimenters. In both cases the conclusions were drawn partly by comparing responses at different points on the response surface.

Production of volatile compounds by Geotrichum fragrans using cassava wastewater as substrate

The cassava processing industry generates wastewater named manipueira with a high organic content. Although considered a pollutant, manipueira can be used as substrate for fermentative processes including the cultivation of Geotrichum fragrans. This aerobic microorganism isolated from cassava wastewater has cyanide resistant respiration. Under cassava wastewater cultivation, G. fragrans produced fruit aroma volatile compounds. This study evaluated volatile compounds produced by G. fragrans in cassava liquid waste. The waste had a sugar composition composed of dextrin (2.6%), maltose (1.4%), sucrose (32.1%), glucose (38.3%), and fructose (25.6%). The average value of total sugars was 58.2 g l(-1), composed of 38.0 g l(-1) reducing and 20.2 g l(-1) non-reducing sugars. The chemical oxygen demand (COD) average value was 60 000 mg l(-1). G. fragrans used sugars (fructose and glucose) for energy generation reducing the COD value of the cassava wastewater by 40%. Biomass production of G. fragrans cultivated for 12 h in natural cassava liquid waste was 12.8 g l(-)1. The volatile compounds identified in the cassava liquid waste after 72 h cultivation were: 1-butanol, 3-methyl 1-butanol (isoamylic alcohol), 2-methyl 1-butanol, 1-3 butanodiol and phenylethanol; ethyl acetate, ethyl propionate, 2-methyl ethyl propionate and 2-methyl propanoic. The effect of substrate supplementation with glucose (50 g l(-1)), fructose (50 g l(-1)) and aqueous yeast extract (200 ml l(-1)) did not affect the qualitative and quantitative profiles of volatile compounds. These results indicate that the carbon (C) source utilized by microorganism was glucose or fructose, while nitrogen (N) supplementation was not necessary because the agent did not exhaust all the nitrogen of the wastewater. (C) 2003 Elsevier B.V. Ltd. All rights reserved.

Concentration profiles and effective diffusion coefficients of sucrose and water in osmo-dehydrated apples

The spatial distribution of water and sugars in half-fresh apples dehydrated in sucrose solutions (30% and 50% w/w, 27 degrees C) for 2, 4 and 8 h, was determined. Each half was sliced as from the exposed surface. The density, water and sugar contents were determined for each piece. A mathematical model was fitted to the experimental data of the water and sucrose contents considering the overall flux and tissue shrinkage. A numerical method of finite differences permitted the calculation of the effective diffusion coefficients as a function of concentration, using material coordinates and integrating the two differential equations (for water and sucrose) simultaneously. The coefficients obtained were one or even two orders of magnitude lower than those for pure solutions and presented unusual concentration dependence. The behaviour of the apple tissue was also studied using light microscopy techniques to obtain images of the osmotically treated pieces (20%, 30% and 50% w/w sucrose solutions for 2, 4 and 8 h). (c) 2006 Elsevier Ltd. All rights reserved.

Peroxidase from peach fruit: Thermal stability

Peroxidase from peach fruit was purified 28.9-fold by DEAE-cellulose, Sephadex G-100 and hydroxylapatite chromatography. The purified enzyme showed only one peak of activity with an optimum pH of 5.0 and temperature of 40 degreesC. The calculated activation energy (Ea) for the reaction was 7.97 kcal/mol. The enzyme was heat-labile in the temperature range of 60 to 80 degreesC with a fast inactivation at 80 degreesC. PAGE of the inactivation course at 70 degreesC showed only one band of activity. Different sugars increased the heat stability of the activity in the following order: sucrose>lactose>glucose>fructose. Measurement of residual activity showed a stabilizing effect of sucrose at various temperature/sugar concentrations (10 to 40%, w/w) with the Ea for inactivation increasing with sucrose concentration from 0 to 20% (w/w). After inactivation at 70 degreesC and 75 degreesC the enzyme was able to be reactivated by up to 40% of the initial activity when stared at 30 degreesC.

Anthracnose control and mango quality (Mangifera indica L.) cv. van dyke after hydrothermal and chemical treatment

Anthracnose causes severe damage to the mango skin. Therefore, there is need to do the post-harvest phytosanitary treatment of the fruits through soaking in fungicide solution, which associated with the hydrothermal treatment has resulted into improved benefits. The present work has aimed at studying anthracnose control and post-harvest quality of mangoes (Mangifera indica L.), cv. Van Dyke after hydrothermal treatment associated with chemical through the physical, physicochemical, chemical and phytopathologic analyses. The mangoes coming from the commercial orchard of the town of Jana ba-MG, of the 2000 crop, were harvested at the commercial maturation stage. After selection according to color uniformity, size and absence of mechanic and physiologic injuries, they were shipped in plastic boxes to the EPAMIG - CTNM - Nova Porteirinha - MG, fruit post-harvest laboratory, where the samples were submitted to the hydrothermal treatment in cold bath in solution containing the fungicides Thiabendazole (Tecto 400 mL/100L), Proclaraz (Sportak 10L/100L) and Imazalil (Magnate 200mL/100L). After air-drying, the fruits were packed in plastic trays and stored at room temperature (25 +/- 2 degrees C and RH 70%) for a 12-day period and evaluated as to the intrinsic quality characteristics every 4 days. The experiment was conducted a completely randomized with 8 treatments, 4 replicates and experimental unit consisting of 4 fruits. The variations of pH, total soluble solids, total titrable acidity and total soluble sugars have not endangered the organoleptic characteristics of mangoes cv. Van Dyke stored under room temperature (25 +/- 2 degrees C and RH 70%) till 8 days' storage. The association of the hydrothermal treatment with the chemical was efficient in fruit anthracnose control for till 12 days' storage. The fungicide Prochloraz (Sportak 110 mL/100L), associated with the hydrothermal treatment, completely inhibited the appearance of anthracnose symptoms.

Reagentless biosensor for isocitrate using one step modified Pt-Ir microelectrode

The Pt-Ir microelectrode modified through one step, electropolymerization is proposed for the isocitrate amperometric biosensor construction. The enzyme (isocitrate dehydrogenase-ICDH), coenzyme (NADP(+)) and mediator (Meldola's Blue) were immobilized onto the microelectrode surface in one step from a PIPES buffer solution containing pyrrole. The optimized experimental conditions were 25 cycles of cyclic voltammetric in a solution containing 3.58 10(-5) mol l(-1) of mediator, 3.51 10(-4) mol l(-1) of coenzyme and 2.68 U ml(-1) of enzyme. In contrast to the biosensor for isocitrate reported in literature, just one enzyme was immobilized and no coenzyme addition in the solution of analysis was necessary. Catalytic currents were proportional to the isocitrate concentration between 7.7 10(-6) and 1.04 10(-4) mol l(-1), showing good repeatability. The detection limit of the proposed biosensor was 3.50 10(-6) mol l(-1), the response time was lower than 20 s, the lifetime was about 30 determinations and no significant interference of sugars and citric acid was verified. Orange juice samples were analysed by both methodology biosensor and spectrophotometric commercial kit, and the obtained results presented a good correlation. The data demonstrated that the developed biosensor is suitable for isocitrate determination in orange juice without matrix interferences. (C) 2001 Elsevier B.V. B.V. All rights reserved.

Flowering phenology, nectary structure and breeding system in Corymborkis flava (Spiranthoideae : Tropidieae), a terrestrial orchid from a Neotropical forest

Flowering phenology, breeding system and nectary structure of Corymborkis flava (Sw.) Kuntze were studied in a fragment of the Atlantic Forest in south-eastern Brazil. The flowering period extended from March (end of rainy season) to early June and seed dispersal occurred from June to September (dry season). Flowering peak occurred mainly in April, with up to 34 open flowers per plant being observed. The yellow, odourless and tubular flowers lasted similar to 7.8 days. The flowers present a perigonal nectary located in the basal lateral parts of the labellum; this is the first report on the nectary location and characterisation in the Tropidieae tribe. At the pre-anthesis stage, cells of both secretory parenchyma and epidermis of the nectary are filled with compound amyloplasts. However, starch grains were not observed in these tissues in senescent flowers, indicating that these starch grains are hydrolysed and used as source of sugars for nectar production. The nectar accumulates between the cuticle and the outer periclinal wall of the epidermal cells before. owing out into the nectar chamber. C. flava is a self-compatible species and spontaneous self-pollination does not occur because of hercogamy. The high pollinia removal (0.80) and insertion (0.82) per flower, as well as the high natural fruit-set indicate an efficient natural pollination system. The present study contributes for the knowledge of the diversity of reproductive strategies and nectary structures in Orchidaceae.

Action of auxins, boron and harvest season on the rooting of kiwi fruit stem cuttings (Actinidia chinensis Pl cv Monty)

Action of auxins on the rooting of stem cuttings of kiwi (Actinidia chinensis P. cv Monty). This work studies the effects of some synthetic auxins and B in the rooting of kiwi (Actinidia chinensis Planch cv Monty) stem cuttings. The treatments used were as follows: T1 (H2O); T2 (NAA 300 ppm); T3 (IBA 300 ppm); T4 (NAA 300 ppm + B); T5 (IBA 300 ppm + B); T6 (NAA 0,5%-talc) and T7 (IBA 0,5%-talc), applied to the bases of the cuttings. These were then placed in rooting dishes with pure vermiculite in a misty nebulization chamber until collection day (120 days). The evaluation of auxin and boric acid effects on kiwi stem cuttings were made based on the following observations: 1. The percentage of rooted stem cuttings; 2. reducing sugars and total sugar analyses (in g/100 g of dry matter); and 3. tryptophan analyses (in mu g/100 mg of dry matter). The results show that summer is the best season for rooting Actinidia chinensis Planch cv Monty stem cuttings. The use of IBA talc 0,5% on the bases of the cuttings shamed positive results too.

Purification and properties of buffalo (Bubalus bubalis) erythrocyte hexokinase

Buffalo erythrocytes contain one isozyme of hexokinase that apparently lacks microheterogeneity as shown by chromatographic properties. A single protein band was detected by means of Western blotting using an antibody raised in rabbits against homogeneous rat brain hexokinase I. The native protein has a molecular weight of 200,000 +/- 2880 by gel filtration. Partial purification of erythrocyte hexokinase by a combination of several procedures, including affinity chromatography, which was previously applied successfully to the purifica tion of other mammalian type I hexokinases, produced a partially purified enzyme that showed several contami nants after SDS-polyacrylamide gel electrophoresis. The affinity of buffalo erythrocyte hexokinase for glucose (K-m = 0.012 +/- 0.001 mM) is lower than most other mammal hexokinases type I. It phosphorylates other sugars, with considerably higher K-m values. This isozyme is able to use MgATP but does not use MgGTP, MgCTP or MgUTP. We used inhibition patterns, obtained with products to elucidate enzyme sequential mechanisms. Our results are clearly in agreement with a random sequential mechanism and in disagreement with an ordered sequential mechanism with either glucose or ATP as the obligatory first substrates. The ADP inhibition was of mixed type with both ATP and glucose as substrates. (C) 1997 Elsevier B.V.