Probing key DNA contacts in AraR-mediated transcriptional repression of the Bacillus subtilis arabinose regulon.

Nucleic Acid Research (2007) Vol.37 N. 14 4755-4766

Síntese e modificações de derivados heterocíclicos de D-arabinose: potenciais inibidores de glicose-6-fosfato isomerase e de glicosamina-6-fosfato sintase

The synthesis of -5-(D-arabino-1,2,3,4-tetrahydroxybutyl)tetrazole and -2-(D-arabino-1,2,3,4-tetra-acetoxybutyl)-5-methyl-1,3,4-oxadiazole from D-arabinose is described. Attempts at removing the protecting groups of the oxadiazole derivative were unsuccessful, leading to products resulting from the opening of the oxadiazole ring. The unprotected tetrazole derivative was selectively phosphorylated at the primary hydroxyl group with diethylphosphoryl chloride. The resulting 5-[D-arabino-4-(diethylphosphoryloxy)-1,2,3-trihydroxybutyl]tetrazole is a protected form of a potential inhibitor of the enzymes glucose-6-phosphate isomerase and glucosamine synthase.

Hydrogenation of L-arabinose, D-galactose, D-maltose and L-rhamnose

Under de senaste åren har intresset för utnyttjandet av förnybara resurser kraftigt ökat. I samband med detta utgör kolhydrater en viktig del av den tillgängliga förnybara biomassan och den har därefter blivit föremål för ett stort intresse inom hållbar kemi. Sockeralkoholer är en särskilt viktig grupp av molekyler som vanligtvis erhålls ur kolhydrater och som har mångsidiga tillämpningar som t.ex. lågkalorihaltiga sötningsmedel. Forskningen i doktorsarbetet omfattar hydreringen av naturligt förekommande sockerarter L-arabinos, D-galaktos, D-maltos och L-ramnos till respektive sockeralkoholer. Dessa sockeralkoholer kan användas bl.a. som hälsosamma sötningsmedel på samma sätt som xylitol. Grunden för detta arbete består av hydreringsexperiment som utfördes på en dispergerad ruteniumkatalysator i syfte att studera bildningskinetiken av de motsvarande sockeralkoholerna. Reaktionerna genomfördes vid temperaturer mellan 90 och 130 °C och vätetryck mellan 40 och 60 bar. Under dessa betingelser var det möjligt att åstadkomma sockeromvandlingar upp till 100 %. Reaktionshastigheterna modellerades matematiskt. Konkurrerande kinetiska modeller som baserades på Langmuir-Hinshelwood-konceptet föreslogs för att beskriva reaktionerna. Parametrar i hastighetsekvationerna bestämdes därefter genom icke-linjär regression. Dessa modeller kunde väl förutsäga hydreringsreaktionernas förlopp och de kan följaktligen användas för design av industriella anläggningar. Ytterligare hydreringsexperiment med sockerblandningar genomfördes för att fördjupa kunskaper i kinetik och reaktionsmekanismer av sockerhydreringen. Studierna genomfördes med syntetiska sockerblandningar av L-arabinos och D-galaktos (de viktigaste komponenterna i hemicellulosan arabinogalaktan). Fullständig omsättning uppnåddes med utmärkta selektiviteter som överskred 95 % och dessutom inverkade varken temperatur eller vätetryck på reaktionens förlopp på något oväntat sätt. Antagandet av konkurrerande adsorption för en samtidig reduktion av båda sockermolekylerna gav en kinetisk modell som noggrant beskrev de experimentella resultaten. Idén om att utforska potentiella sätt att påskynda bildningen av sockeralkoholer ledde till utföringen av hydreringsexperiment med L-arabinos och D-galaktos i närvaro av ultraljud. Det visade sig att ultraljudets inverkan var oberoende av sockerhalten och vätetrycket och att bestrålningen gynnade hydreringen av D-galaktos trots att den inte förhindrade Ru/C-katalysatorns deaktivering överhuvudtaget. En kinetisk modell som beaktade deaktiveringen utvecklades. Kontinuerlig hydrering av L-arabinos genomfördes med tre olika Ru-katalysatorer på tre olika bärare: tyg av aktivt kol, kolnanorör på svampliknande metalliska strukturer samt krossade partiklar av en kommersiell Ru/C-katalysator. Det visade sig att det var möjligt att omvandla L-arabinos till arabitol med höga selektiviteter med hjälp av Ru/koltyg-katalysatorn. Dessa experiment demonstrerade att hydreringen av de valda sockerarterna är en helt genomförbar rutt till framställning av fin- och specialkemikalier, som kan förverkligas i större skala.

Solubility and Seeded Metastable Zone width of functional sugar L-arabinose

L-arabinose is widely used in food, medicine, chemistry, and biology fields; however, solubility and seeded metastable zone width (MSZW) of L-arabinose have not been reported in the literature. In this paper, solubility and MSZW of L-arabinose in aqueous solution were determined. Solubility of L-arabinose was measured in the range of 20-68 °C by a conventional equilibrium solubility method and quantitation was determined using the ion chromatography technique. Seeded MSZW was determined in the range of 51-73% by the calorimetric method. The effect of two salts (potassium chloride and calcium chloride) on the solubility and MSZW of L-arabinose were also evaluated. Results showed that both potassium chloride and calcium chloride increased the solubility of L-arabinose, and this increase was intensified with temperature rise. The MSZW of L-arabinose was not constant but a spread. Potassium chloride increased the MSZW of L-arabinose. However, the effect of calcium chloride on MSZW of L-arabinose was concentration dependent. Conclusion: the L-arabinose solubility increased with the increase in temperature, and both potassium chloride and calcium chloride increased the solubility of L-arabinose in aqueous solution. The seeded MSZW of L-arabinose is not a constant; it increases in the presence of potassium chloride and varies with the change in calcium chloride concentration.

Arabinose and protocatechuate catabolism genes are important for growth of Rhizobium leguminosarum biovar viciae in the pea rhizosphere

Background and aims: To form nitrogen-fixing nodules on pea roots, Rhizobium leguminosarum biovar viciae must be competitive in the rhizosphere. Our aim was to identify genes important for rhizosphere fitness. Methods: Signature-tagged mutants were screened using microarrays to identify mutants reduced for growth in pea rhizospheres. Candidate mutants were assessed relative to controls for growth in minimal medium, growth in pea rhizospheres and for infection of peas in mixed inoculants. Mutated genes were identified by DNA sequencing and confirmed by transduction. Results: Of 5508 signature-tagged mutants, microarrays implicated 50 as having decreased rhizosphere fitness. Growth tests identified six mutants with rhizosphere-specific phenotypes. The mutation in one of the genes (araE) was in an arabinose catabolism operon and blocked growth on arabinose. The mutation in another gene (pcaM), encoding a predicted solute binding protein for protocatechuate and hydroxybenzoate uptake, decreased growth on protocatechuate. Both mutants were decreased for nodule infection competitiveness with mixed inoculants, but nodulated peas normally when inoculated alone. Other mutants with similar phenotypes had mutations predicted to affect secondary metabolism. Conclusions: Catabolism of arabinose and protocatechuate in the pea rhizosphere is important for competitiveness of R.l. viciae. Other genes predicted to be involved in secondary metabolism are also important.

Seleção de leveduras fermentadoras de D-xilose / L-arabinose

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A study on chemical constituents and sugars extraction from spent coffee grounds

Spent coffee grounds (SCG) the residual materials obtained during the processing of raw coffee powder to prepare instant coffee are the main coffee Industry residues In the present work this material was chemically characterized and subsequently submitted to a dilute acid hydrolysis aiming to recover the hemicellulose sugars Reactions were performed according to experimental designs to verify the effects of the variables H(2)SO(4) concentration liquid-to-solid ratio temperature and reaction time on the efficiency of hydrolysis SCG was found to be rich in sugars (45 3% w/w) among of which hemicellulose (constituted by mannose galactose and arabinose) and cellulose (glucose homopolymer) correspond to 367% (w/w) and 8 6% (w/w) respectively Optimal conditions for hemicellulose sugars extraction consisted in using 100 mg acid/g dry matter 10g liquid/g solid at 163 degrees C for 45 min Under these conditions hydrolysis efficiencies of 100% 774% and 895% may be achieved for galactan mannan and arabinan respectively corresponding to a hemicellulose hydrolysis efficiency of 874% (C) 2010 Elsevier Ltd All rights reserved

Xylooligosaccharides Production from Alkali-Pretreated Sugarcane Bagasse Using Xylanases from Thermoascus aurantiacus

Sugarcane bagasse hemicellulose was isolated in a one-step chemical extraction using hydrogen peroxide in alkaline media. The polysaccharide containing 80.9% xylose and small amounts of L-arabinose, 4-O-methyl-D-glucuronic acid and glucose, was hydrolyzed by crude enzymatic extracts from Thermoascus aurantiacus at 50 degrees C. Conditions of enzymatic hydrolysis leading to the best yields of xylose and xylooligosaccharides (DP 2-5) were investigated using substrate concentration in the range 0.5-3.5% (w/v), enzyme load 40-80 U/g of the substrate, and reaction time from 3 to 96 h, applying a 22 factorial design. The maximum conversion to xylooligosaccharides (37.1%) was obtained with 2.6% of substrate and xylanase load of 60 U/g. The predicted maximum yield of xylobiose by a polynomial model was 41.6%. Crude enzymatic extract of T. aurantiacus generate from sugarcane bagasse hemicellulose 39% of xylose, 59% of xylobiose, and 2% of other xylooligosaccharides.

Chemical composition changes in eucalyptus and pinus woods submitted to heat treatment

This study investigated the influence of heat treatment on the chemical composition of Eucalyptus saligna and Pinus caribaea var. hondurensis woods to understand its role in wood processing. E. saligna and P. caribaea var. hondurensis woods were treated in a laboratorial electric furnace at 120, 140, 160 and 180 degrees C to induce their heat treatment. The chemical composition of the resulting products and those from original wood were determined by gas chromatography. Eucalyptus and Pinus showed a significant reduction in arabinose, manose, galactose and xylose contents when submitted to increasing temperatures. No significant alteration in glucose content was observed. Lignin content, however, increased during the heat process. There was a significant reduction in extractive content for Eucalyptus. On the other hand, a slight increase in extractive content has been determined for the Pinus wood. and that only for the highest temperature. These different behaviors can be explained by differences in chemical constituents between softwoods and hardwoods. The results obtained in this study provide important information for future research and utilization of thermally modified wood. (c) 2008 Elsevier Ltd. All rights reserved.

Effect of cooking on non-starch polysaccharides of hard-to-cook beans

Experiments carried out to study changes induced by hard-to-cook (HTC) phenomenon in the non-starch polysaccharides of beans stored at 30 degrees C and 75% RH for 8 months showed that the development of HTC did not affect the amounts of soluble and insoluble fibre in cooked seeds but changed their carbohydrates physical properties. Aged beans non-starch polysaccharides presented lower water-solubility and underwent lower degradation of galacturonans and arabinose-rich polysaccharides when submitted to cooking. The decrease in non-starch polysaccharides water-solubility produced a shift in the polymers fractionation profile which resulted in an increase of weak and middle-alkali soluble polymers bulk as well as in their arabinose and uronic acid contents. Uronic acid contents were higher in polymers released by 1 M NaOH and in the cellulose-rich residues while the arabinose contents were higher in the mild-alkali soluble polymers of aged seeds. Methylation analysis showed no evident alterations in the xyloglucans and arabinans branching degree with beans ageing. However, both, the molecular mass of water-soluble pectins and CDTA-soluble pectins, increased. Even though changes in the non-starch polysaccharide solubility were not related to the decrease in the arabinan and xyloglucan degree of branching they may be related to the formation of new chemical interactions other than hydrogen bond. There was a correlation between acidic and neutral polysaccharides insolubilisation in beans ageing and probably in beans hardening. After processing, aged seeds present higher amounts of insoluble fibre when compared to normal beans. (C) 2008 Elsevier Ltd. All rights reserved.

Superprodução, purificação e caracterização parcial de complexos proteína-polissacáridos inovadores a partir de estirpes de basidiomicetos

O presente trabalho teve como objectivo o estudo de meios de cultura, tendo em vista a superprodução de complexos proteína-polissacáridos, intracelulares e extracelulares, a partir dos basidiomicetos Pleurotus ostreatus e Lentinula edodes. O meio de cultura suplementado com soro de leite apresentou diversas vantagens para a obtenção dessas macromoléculas biológicas, face aos restantes meios analisados, pelo que foi seleccionado para a produção em fermentador de laboratório em regime descontínuo. O crescimento da cultura foi efectuado a 28°C, 200 rpm e durante 10 dias, e as produtividades de complexos proteína-polissacáridos obtidas para o Pleurotus ostreatus foram de 0,008 ± 8,12 x 10-5 g/(L.dia) e de 0,324 ± 0,005 g/(L.dia), para os complexos precipitados a partir da biomassa e do caldo de fermentação, respectivamente. Posteriormente, os complexos intracelulares e extracelulares foram purificados por cromatografia de filtração em gel e parcialmente caracterizados, revelando concentrações de polissacáridos superiores à concentração proteica, pseudo-actividade de superóxido dismutase, e valores de Mr entre 64 e 9 000kDa. As amostras de complexos proteína-polissacáridos foram caracterizadas por HPLC e apresentaram picos de UV e IR com tempos de retenção de aproximadamente 6 e 12 minutos, respectivamente. Após hidrólise ácida, os complexos heteropolissacáridos obtidos apresentaram na sua constituição glucose, ramnose e arabinose. Adicionalmente, a análise dos complexos proteína-polissacáridos por FT-IR revelou bandas de absorção características destas macromoleculas biológicas, designadamente, a 846,5; 1032,3; 1186,4; 1471,0; 1648,8; 2739,6 e 3419,3 cm-1 para o Pleurotus ostreatus e 862,3; 1044,1; 1474,9; 1644,8; 2372,0; 2992,5; e 3415,4 cm-1 para Lentinula edodes.

Ocorrência e diversidade de leveduras no filoplano de plantas seleccionadas do Parque Natural da Serra da Arrábida

Dissertação apresentada para obtenção do Grau de Doutor em Biologia (especialidade Microbiologia), pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia

Hexose and pentose transport inascomycetousyeasts: an overview

FEMS Yeast Research, Vol. 9, nº 4

A multitask ATPase serving different ABC-type sugar importers in Bacillus subtilis

Journal of Bacteriology (Out 2010) 5312-5318

Two distinct arabinofuranosidases contribute to arabino-oligosaccharide degradation in Bacillus subtilis.

Microbiology 154 (2008) 2719-2729