Efeito do resfriamento sobre açúcares solúveis, taxa de respiração, fenóis totais, atividade de peroxidase e dormência de bulbos de cebola

Além de ser uma das hortaliças mais cultivada e consumida, bulbos de cebola são afetados, durante a armazenagem, por fatores fisiológicos, bioquímicos e tecnológicos, que podem afetar seus atributos de qualidade. Taxa de respiração (RR O2), açucares solúveis (SS), fenóis totais (TP) e atividade de peroxidase (POD) foram medidos em tecidos internos de brotos, durante uma quebra de dormência de bulbos de cebola, tratados por quatro semanas a 0ºC e armazenados no escuro a 20ºC. Bulbos controle foram armazenados simultaneamente na mesma condição. A quebra da dormência foi verificada através do aparecimento das primeiras folhas internas verdes, cortando 30 bulbos longitudinalmente. Depois de oito semanas a RR O2 de bulbos brotados foi 52% maior em relação a bulbos recentemente colhidos e bulbos dormentes. Os SS diminuíram uma semana após resfriamento, de 15 para 9 mg g-1 de peso fresco e depois apresentaram um pico, de 9 para 19 mg g-1 depois de três semanas. Para os bulbos controle também foi observado um pico similar depois de seis semanas. Para os brotos internos de cebolas tratadas com frio, foi observado um pequeno aumento de TP (de 0,17 a 0,2 mg g-1 de peso fresco), durante as duas primeiras semanas de resfriamento e, depois, um decréscimo para 0,11 mg g-1 depois de oito semanas. Para os brotos internos de bulbos controle, os TP também aumentaram pouco, de 0,17 para 0,2 mg g-1 depois de cinco semanas, decrescendo para 0,15 mg g-1 depois de sete semanas, quando começaram a brotar. A atividade de POD apresentou uma tendência similar em relação aos TP. Para bulbos tratados a frio, a atividade POD aumentou para 1,7 U g-1 de peso fresco depois de duas semanas, decrescendo para 1,1 U g-1 durante as últimas quatro semanas. Para as amostras controle, a atividade POD permaneceu estável por quatro semanas e decresceu progressivamente de 29% durante as últimas quatro semanas. O decréscimo da atividade de POD coincidiu com o decréscimo de TP e coincidiu com o início da brotação. Com o tratamento frio, os primeiros brotos foram observados durante a terceira semana, enquanto o brotamento total se deu depois de oito semanas. Em comparação, apenas 20% dos bulbos controle brotaram depois de oito semanas.Besides onions being one of the most cultivated and consumed vegetables, during storage onion bulbs are still affected by many physiological, biochemical and technological factors which can influence their quality. Respiration rate (RR O2), soluble sugars (SS), total phenolics (TP), and peroxidase (POD) activity were measured in inner bud tissues during a dormancy break of onion bulbs treated four weeks at 0ºC and stored in the dark at 20ºC. Control bulbs were stored simultaneously in the same condition. Breakage of dormancy was checked by the appearance of first green internal leaves by cutting longitudinally 30 bulbs. After eight weeks, RR O2 of sprouted onions was 52% higher than that of freshly harvested and dormant bulbs. One week after cooling SS decreased from 15 to 9 mg g-1 fresh weight, and then peaked from 9 to 19 mg g-1 after three weeks. For control bulbs, a similar peak was observed after six weeks. For inner buds of cold-treated onions, a slight increase of TP (from 0.17 to 0.2 mg g-1; fresh weight) was observed during the first two weeks of cooling, and then a decrease to 0.11 mg g-1 was observed after eight weeks. For inner buds of control bulbs, TP also increased slightly from 0.17 to 0.2 mg g-1 after five weeks, and decreased to 0.15 mg g-1 after seven weeks when bulbs began to sprout. POD activity showed a similar pattern in relation to TP. For cold-treated bulbs, POD activity increased to 1.7 U g-1 fresh weight after two weeks, and decreased to 1.1 U g-1 during the last four weeks. For control samples, POD activity was stable during 4 weeks and decreased progressively by 29% during the last four weeks. This decrease in POD activity coincided with the decrease in TP, and coincided with onset of sprouting. With cold treatment, first sprouts were observed during the third week, while total sprouting was observed after eight weeks. In comparison, only 20% of the control bulbs sprouted after the period of 8 weeks

Cloning and functional characterization of a fructan 1-exohydrolase (1-FEH) in edible burdock (Arctium lappa L.)

<p>Abstract</p> <p>Background</p> <p>We have previously reported on the variation of total fructooligosaccharides (FOS), total inulooligosaccharides (IOS) and inulin in the roots of burdock stored at different temperatures. During storage at 0°C, an increase of FOS as a result of the hydrolysis of inulin was observed. Moreover, we suggested that an increase of IOS would likely be due to the synthesis of the IOS by fructosyltransfer from 1-kestose to accumulated fructose and elongated fructose oligomers which can act as acceptors for fructan:fructan 1-fructosyltransferase (1-FFT). However, enzymes such as inulinase or fructan 1-exohydorolase (1-FEH) involved in inulin degradation in burdock roots are still not known. Here, we report the isolation and functional analysis of a gene encoding burdock 1-FEH.</p> <p>Results</p> <p>A cDNA, named <it>aleh1</it>, was obtained by the RACE method following PCR with degenerate primers designed based on amino-acid sequences of FEHs from other plants. The <it>aleh1 </it>encoded a polypeptide of 581 amino acids. The relative molecular mass and isoelectric point (<it>pI</it>) of the deduced polypeptide were calculated to be 65,666 and 4.86. A recombinant protein of <it>aleh1 </it>was produced in <it>Pichia pastoris</it>, and was purified by ion exchange chromatography with DEAE-Sepharose CL-6B, hydrophobic chromatography with Toyopearl HW55S and gel filtration chromatography with Toyopearl HW55S. Purified recombinant protein showed hydrolyzing activity against β-2, 1 type fructans such as 1-kestose, nystose, fructosylnystose and inulin. On the other hand, sucrose, neokestose, 6-kestose and high DP levan were poor substrates.</p> <p>The purified recombinant protein released fructose from sugars extracted from burdock roots. These results indicated that <it>aleh1 </it>encoded 1-FEH.</p

Three novel oligosaccharides synthesized using Thermoanaerobacter brockii kojibiose phosphorylase

<p>Abstract</p> <p>Background</p> <p>Recently synthesized novel oligosaccharides have been produced primarily by hydrolases and glycosyltransferases, while phosphorylases have also been subject of few studies. Indeed, phosphorylases are expected to give good results via their reversible reaction. The purpose of this study was to synthesis other novel oligosaccharides using kojibiose phosphorylase.</p> <p>Results</p> <p>Three novel oligosaccharides were synthesized by glucosyltransfer from β-D-glucose 1-phosphate (β-D-G1P) to xylosylfructoside [<it>O</it>-α-D-xylopyranosyl-(1→2)-β-D-fructofuranoside] using <it>Thermoanaerobacter brockii </it>kojibiose phosphorylase. These oligosaccharides were isolated using carbon-Celite column chromatography and preparative high performance liquid chromatography. Gas liquid chromatography analysis of methyl derivatives, MALDI-TOF MS and NMR measurements were used for structural characterisation. The ¹H and ¹³C NMR signals of each saccharide were assigned using 2D-NMR including COSY (correlated spectroscopy), HSQC (herteronuclear single quantum coherence), CH₂-selected E-HSQC (CH₂-selected Editing-HSQC), HSQC-TOCSY (HSQC-total correlation spectroscopy) and HMBC (heteronuclear multiple bond correlation).</p> <p>Conclusion</p> <p>The structure of three synthesized saccharides were determined, and these oligosaccharides have been identified as <it>O</it>-α-D-glucopyranosyl-(1→2)-<it>O</it>-α-D-xylopyranosyl-(1→2)-β-D-fructofuranoside (saccharide <b>1</b>), <it>O</it>-α-D-glucopyranosyl-(1→2)-<it>O</it>-α-D-glucopyranosyl-(1→2)-<it>O</it>-α-D-xylopyranosyl-(1→2)-β-D-fructofuranoside (saccharide <b>2</b>) and <it>O</it>-α-D-glucopyranosyl-(1→[2-<it>O</it>-α-D-glucopyranosyl-1]₂→2)-<it>O</it>-α-D-xylopyranosyl-(1→2)-β-D-fructofuranoside (saccharide <b>3</b>).</p

Characterization of recombinant β-fructofuranosidase from Bifidobacterium adolescentis G1

<p>Abstract</p> <p>Background</p> <p>We have previously reported on purification and characterization of β-fructofuranosidase (β-FFase) from <it>Bifidobacterium adolescentis </it>G1. This enzyme showed high activity of hydrolysis on fructo-oligosaccharides with a low degree of polymerization. Recently, genome sequences of <it>B. longum </it>NCC2705 and <it>B. adolescentis </it>ATCC 15703 were determined, and <it>cscA </it>gene in the both genome sequences encoding β-FFase was predicted. Here, cloning of <it>cscA </it>gene encoding putative β-FFase from <it>B. adolescentis </it>G1, its expression in <it>E. coli </it>and properties of the recombinant protein are described.</p> <p>Results</p> <p>Using the information of <it>cscA </it>gene from <it>Bifidobacterium adolescentis </it>ATCC 15703, <it>cscA </it>gene from <it>B. adolescentis </it>G1 was cloned and sequenced. The N-terminal amino acid sequence of purified β-FFase from <it>B. adolescentis </it>G1 was identical to the deduced amino acid sequences of <it>cscA </it>gene from <it>B. adolescentis </it>G1. To confirm the translated product of the <it>cscA </it>gene, the recombinant protein was expressed in <it>Escherichia coli</it>. Molecular mass of the purified recombinant enzyme was estimated to be about 66,000 by SDS-PAGE and 60,300 by MALDI TOF-MS. The optimum pH of the enzyme was 5.7 and the enzyme was stable at pH 5.0-8.6. The thermostability of the enzyme was up to 50°C. The <it>K</it>_m(mM), <it>V</it>_max(μmol/mg of protein/min), <it>k</it>₀(sec^-1) and <it>k</it>₀/<it>K</it>_m(mM^-1sec^-1) for 1-kestose, neokestose, nystose, fructosylnystose, sucrose and inulin were 1.7, 107, 107.5, 63.2, and 1.7, 142, 142.7, 83.9, and 3.9, 152, 152.8, 39.2, and 2.2, 75, 75.4, 34.3, and 38, 79, 79.4, 2.1, and 25.9, 77, 77.4, 3.0, respectively. The hydrolytic activity was strongly inhibited by AgNO₃, SDS, and HgCl₂.</p> <p>Conclusion</p> <p>The recombinant enzyme had similar specificity to the native enzyme, high affinity for 1-kestose, and low affinity for sucrose and inulin, although properties of the recombinant enzyme showed slight difference from those of the native one previously described.</p

構造研究に基づく南海トラフ（西部）地震発生帯のプレート形状および速度構造の3次元モデル

Great interplate earthquakes have repeatedly occurred in pairs along the Nankai Trough. In order to reduce a great deal of damage to coastal area from both strong ground motion and tsunami generation, it is necessary to understand rupture synchronization and segmentation of the Nankai megathrust earthquake. For a precise estimate of the rupture zone of the Nankai megathrust event based on the knowledge of realistic earthquake cycles and variations of magnitude, it is important to know the geometry and property of the plate boundary of the subduction seismogenic zone. To improve a physical model of the Nankai Trough seismogenic zone, the large-scale high-resolution wide-angle and reflection (MCS) seismic studies, and long-term observation have been conducted since 2008. Marine active source seismic data have been acquired along grid two-dimensional profiles having the total length of ~800km per year. A three-dimensional seismic tomography using active and passive seismic data observed both land and ocean bottom stations have been also performed. This study is part of 'Research concerning Interaction Between the Tokai, Tonankai and Nankai Earthquakes' funded by Ministry of Education, Culture, Sports, Science and Technology, Japan. The seismic survey was conducted off the Tokai area including the onshore survey across the eastern Kii Peninsula in 2012, the final year of this project. Compiling those studies provides a three-dimensional plate geometry and velocity structure models of the western Nankai Trough at the moment. Although their reliability and resolution should be evaluated, these models can be applied to a numerical simulation to examine if the observed rupture zone of the historical event can be reproduced. We will also try to construct more fine-scale model for the entire Nankai Trough area.SSS31-P15ポスター要旨 / 日本地球惑星科学連合2013年大会（2013年5月19日～5月24日, 幕張メッセ国際会議場） / 日本惑星科学連合の許諾に基づき本文ファイルを掲載http://www.godac.jamstec.go.jp/darwin/cruise/kairei/kr10-11/ehttp://www.godac.jamstec.go.jp/darwin/cruise/kairei/kr11-09/

Chemical Structure and Localization of Levan, the Predominant Fructan Type in Underground Systems of Gomphrena marginata (Amaranthaceae)

Gomphrena marginata Seub. (Amaranthaceae) is an endemic species from Brazilian campos rupestres with a fructan accumulating underground reserve system. Analyses of high performance anion exchange chromatography (HPAEC–PAD) revealed the presence of the soluble carbohydrates glucose, fructose, sucrose, 1-kestose, 6-kestose, nystose and fructans with degree of polymerization (DP) up to approximately 40 fructose units. Data of 1H and 13C Nuclear Magnetic Resonance (NMR) spectroscopy, including Heteronuclear Single-Quantum Correlation (HSQC) and Heteronuclear Multiple-Bonds Correlation (HMBC) showed the presence of β (2,6) linkages, characteristic of the linear molecule of levan-type fructan(2,6). These results confirmed previous studies suggesting that the reserve carbohydrate in the underground system of this species was levan-type fructans, similar to that of G. macrocephala. Structural analyses of the thickened underground system using light microscopy revealed a mixed origin system consisting mainly of a gemmiferous tuberous root with the upper region formed by short branched stems, both presenting vascular cylinders with unusual growth patterns. Fructan spherocrystals were visualized under polarized light and scanning electron microscopy (SEM) mostly in the cortex and vascular cylinder in both thickened stem and root. In addition to data reported in the literature concerning the occurrence of fructans in the Amaranthaceae, the results presented here suggest that fructans are a trait in this family while the levan-type fructan prevail in Gomphrena species

Structural analysis of three novel trisaccharides isolated from the fermented beverage of plant extracts

<p>Abstract</p> <p>Background</p> <p>A fermented beverage of plant extracts was prepared from about fifty kinds of vegetables and fruits. Natural fermentation was carried out mainly by lactic acid bacteria (<it>Leuconostoc </it>spp.) and yeast (<it>Zygosaccharomyces </it>spp. and <it>Pichia </it>spp.). We have previously examined the preparation of novel four trisaccharides from the beverage: <it>O</it>-β-D-fructopyranosyl-(2->6)-<it>O</it>-β-D-glucopyranosyl-(1->3)-D-glucopyranose, <it>O</it>-β-D-fructopyranosyl-(2->6)-<it>O</it>-[β-D-glucopyranosyl-(1->3)]-D-glucopyranose, <it>O</it>-β-D-glucopyranosyl-(1->1)-<it>O</it>-β-D-fructofuranosyl-(2<->1)-α-D-glucopyranoside and <it>O</it>-β-D-galactopyranosyl-(1->1)-<it>O</it>-β-D-fructofuranosyl-(2<->1)- α-D-glucopyranoside.</p> <p>Results</p> <p>Three further novel oligosaccharides have been found from this beverage and isolated from the beverage using carbon-Celite column chromatography and preparative high performance liquid chromatography. Structural confirmation of the saccharides was provided by methylation analysis, MALDI-TOF-MS and NMR measurements.</p> <p>Conclusion</p> <p>The following novel trisaccharides were identified: <it>O</it>-β-D-fructofuranosyl-(2->1)-<it>O</it>-[β-D-glucopyranosyl-(1->3)]-β-D-glucopyranoside (named "3^G-β-D-glucopyranosyl β, β-isosucrose"), <it>O</it>-β-D-glucopyranosyl-(1->2)-<it>O</it>-[β-D-glucopyranosyl-(1->4)]-D-glucopyranose (4¹-β-D-glucopyranosyl sophorose) and <it>O</it>-β-D-fructofuranosyl-(2->6)-<it>O</it>-β-D-glucopyranosyl-(1->3)-D-glucopyranose (6²-β-D-fructofuranosyl laminaribiose).</p

Initial Surgical Versus Conservative Strategies in Patients With Asymptomatic Severe Aortic Stenosis

AbstractBackgroundCurrent guidelines generally recommend watchful waiting until symptoms emerge for aortic valve replacement (AVR) in asymptomatic patients with severe aortic stenosis (AS).ObjectivesThe study sought to compare the long-term outcomes of initial AVR versus conservative strategies following the diagnosis of asymptomatic severe AS.MethodsWe used data from a large multicenter registry enrolling 3,815 consecutive patients with severe AS (peak aortic jet velocity >4.0 m/s, or mean aortic pressure gradient >40 mm Hg, or aortic valve area <1.0 cm2) between January 2003 and December 2011. Among 1,808 asymptomatic patients, the initial AVR and conservative strategies were chosen in 291 patients, and 1,517 patients, respectively. Median follow-up was 1,361 days with 90% follow-up rate at 2 years. The propensity score–matched cohort of 582 patients (n = 291 in each group) was developed as the main analysis set for the current report.ResultsBaseline characteristics of the propensity score–matched cohort were largely comparable, except for the slightly younger age and the greater AS severity in the initial AVR group. In the conservative group, AVR was performed in 41% of patients during follow-up. The cumulative 5-year incidences of all-cause death and heart failure hospitalization were significantly lower in the initial AVR group than in the conservative group (15.4% vs. 26.4%, p = 0.009; 3.8% vs. 19.9%, p < 0.001, respectively).ConclusionsThe long-term outcome of asymptomatic patients with severe AS was dismal when managed conservatively in this real-world analysis and might be substantially improved by an initial AVR strategy. (Contemporary Outcomes After Surgery and Medical Treatment in Patients With Severe Aortic Stenosis Registry; UMIN000012140

昭和南海地震震源域-四国沖～紀伊半島沖-の構造変化

BE12-54講演要旨 / ブルーアース2012（2012年3月22日～23日, 東京海洋大学品川キャンパス