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Valeria Monteze Guimaraes - One of the best experts on this subject based on the ideXlab platform.
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purification and characterization of aspergillus terreus α galactosidases and their use for hydrolysis of soymilk oligosaccharides
Applied Biochemistry and Biotechnology, 2011Co-Authors: Joana Gasperazzo Ferreira, Valeria Monteze Guimaraes, Lilian Da Silva Fialho, Angelica Pataro Reis, Daniel Luciano Falkoski, Sebastiao Tavares De RezendeAbstract:α-Galactosidases has the potential to hydrolyze α-1-6 linkages in raffinose family oligosaccharides (RFO). Aspergillus terreus cells cultivated on wheat bran produced three extracellular forms of α-galactosidases (E1, E2, and E3). E1 and E2 α-galactosidases presented maximal activities at pH 5, while E3 α-galactosidase was more active at pH 5.5. The E1 and E2 enzymes showed stability for 6 h at pH 4–7. Maximal activities were determined at 60, 55, and 50°C, for E1, E2, and E3 α-galactosidase, respectively. E2 α-galactosidase retained 90% of its initial activity after 70 h at 50°C. The enzymes hydrolyzed ρNPGal, melibiose, raffinose and Stachyose, and E1 and E2 enzymes were able to hydrolyze guar gum and locust bean gum substrates. E1 and E3 α-galactosidases were completely inhibited by Hg2+, Ag+, and Cu2+. The treatment of RFO present in soy milk with the enzymes showed that E1 α-galactosidase reduced the Stachyose content to zero after 12 h of reaction, while E2 promoted total hydrolysis of raffinose. The complete removal of the oligosaccharides in soy milk could be reached by synergistic action of both enzymes
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debaryomyces hansenii ufv 1 intracellular α galactosidase characterization and comparative studies with the extracellular enzyme
Journal of Agricultural and Food Chemistry, 2009Co-Authors: Pollyanna Amaral Viana, Sebastiao Tavares De Rezende, Flavia Maria Lopes Passos, Marcelo P Bemquerer, Jamil S Oliveira, Kadima N Teixeira, Alexandre Martins Costa Santos, Jose C Rosa, Marcelo M Santoro, Valeria Monteze GuimaraesAbstract:Debaryomyces hansenii cells cultivated on galactose produced extracellular and intracellular alpha-galactosidases, which showed 54.5 and 54.8 kDa molecular mass (MALDI-TOF), 60 and 61 kDa (SDS-PAGE) and 5.15 and 4.15 pI values, respectively. The extracellular and intracellular deglycosylated forms presented 36 and 40 kDa molecular mass, with 40 and 34% carbohydrate content, respectively. The N-terminal sequences of the alpha-galactosidases were identical. Intracellular alpha-galactosidase showed smaller thermostability when compared to the extracellular enzyme. D. hansenii UFV-1 extracellular alpha-galactosidase presented higher kcat than the intracellular enzyme (7.16 vs 3.29 s-1, respectively) for the p-nitrophenyl-alpha-D-galactopyranoside substrate. The Km for hydrolysis of pNPalphaGal, melibiose, Stachyose, and raffinose were 0.32, 2.12, 10.8, and 32.8 mM, respectively. The intracellular enzyme was a competitively inhibited by galactose (Ki = 0.70 mM), and it was inactivated by Cu(II) and Ag(I). Enzyme incubation with soy milk for 6 h at 55 degrees C reduced Stachyose and raffinose amounts by 100 and 73%, respectively.
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characterization and biotechnological application of an acid α galactosidase from tachigali multijuga benth seeds
Phytochemistry, 2008Co-Authors: Lilian Da Silva Fialho, Valeria Monteze Guimaraes, Carina Marin Callegari, Angelica Pataro Reis, Daianny Silveira Barbosa, Eduardo Euclydes De Lima E Borges, Maurilio Alves Moreira, Sebastiao Tavares De RezendeAbstract:Abstract Tachigali multijuga Benth. seeds were found to contain protein (364 mg g−1 dwt), lipids (24 mg g−1 dwt), ash (35 mg g−1 dwt), and carbohydrates (577 mg g−1 dwt). Sucrose, raffinose, and Stachyose concentrations were 8.3, 3.0, and 11.6 mg g−1 dwt, respectively. α-Galactosidase activity increased during seed germination and reached a maximum level at 108 h after seed imbibition. The α-galactosidase purified from germinating seeds had an Mr of 38,000 and maximal activity at pH 5.0–5.5 and 50 °C. The enzyme was stable at 35 °C and 40 °C, but lost 79% of its activity after 30 min at 50 °C. The activation energy (Ea) values for p-nitrophenyl-α- d -galactopyranoside (pNPGal) and raffinose were 13.86 and 4.75 kcal mol−1, respectively. The Km values for pNPGal, melibiose, raffinose, and Stachyose were 0.45, 5.37, 39.62 and 48.80 mM, respectively. The enzyme was sensitive to inhibition by HgCl2, SDS, AgNO3, CuSO4, and melibiose. d -Galactose was a competitive inhibitor (Ki = 2.74 mM). In addition to its ability to hydrolyze raffinose and Stachyose, the enzyme also hydrolyzed galactomannan.
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hydrolysis of oligosaccharides in soybean products by debaryomyces hansenii ufv 1 α galactosidases
Food Chemistry, 2007Co-Authors: Pollyanna Amaral Viana, Sebastiao Tavares De Rezende, Maurilio Alves Moreira, Daniel Luciano Falkoski, Thiago De Almeida Leite, Ines Charnel Jose, Valeria Monteze GuimaraesAbstract:α-Galactosides are abundant sugars found in legumes such as soybean. Since humans and monogastric animals lack α-galactosidase in the digestive tract, they are unable to digest these sugars, which induce flatulence. The use of α-galactosidases is promising as a means to overcome this problem, and to increase the consumption of soy products. Immobilized α-galactosidase, derived from Debaryomyces hansenii UFV-1, exhibited an activity of 40 U per g of silica and an activity yield of 50%. The optimum pH of free and immobilized α-galactosidase was 5.0 and the optima temperatures were 60 and 80 °C, respectively. The soymilk Stachyose was completely hydrolyzed by different enzyme forms after incubation for 4 h at 60 °C, while raffinose was reduced by 100%, 25% and 68% by free, immobilized enzymes and permeabilized cells, respectively. The soy molasses treatment with free enzyme for 6 h promoted reduction in Stachyose and raffinose contents by 100% and 50%, respectively.
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extracellular α galactosidase from debaryomyces hansenii ufv 1 and its use in the hydrolysis of raffinose oligosaccharides
Journal of Agricultural and Food Chemistry, 2006Co-Authors: Pollyanna Amaral Viana, Sebastiao Tavares De Rezende, Virginia M Marques, Larissa Mattos Trevizano, Flavia Maria Lopes Passos, Maria Goreti De Almeida Oliveira, Marcelo P Bemquerer, Jamil S Oliveira, Valeria Monteze GuimaraesAbstract:Raffinose oligosaccharides (RO) are the factors primarily responsible for flatulence upon ingestion of soybean-derived products. ROs are hydrolyzed by α-galactosidases that cleave α-1,6-linkages of α-galactoside residues. The objectives of this study were the purification and characterization of extracellular α-galactosidase from Debaryomyces hansenii UFV-1. The enzyme purified by gel filtration and anion exchange chromatographies presented an Mr value of 60 kDa and the N-terminal amino acid sequence YENGLNLVPQMGWN. The Km values for hydrolysis of pNPαGal, melibiose, Stachyose, and raffinose were 0.30, 2.01, 9.66, and 16 mM, respectively. The α-galactosidase presented absolute specificity for galactose in the α-position, hydrolyzing pNPGal, Stachyose, raffinose, melibiose, and polymers. The enzyme was noncompetitively inhibited by galactose (Ki = 2.7 mM) and melibiose (Ki = 1.2 mM). Enzyme treatments of soy milk for 4 h at 60 °C reduced the amounts of Stachyose and raffinose by 100%. Keywords: α-Galactosi...
Ralph L. Obendorf - One of the best experts on this subject based on the ideXlab platform.
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Cyclitol galactosides in low-raffinose, low-Stachyose soybean embryos after feeding d-chiro-inositol, myo-inositol or d-pinitol
Seed Science Research, 2013Co-Authors: Elizabeth M. Sensenig, Ralph L. Obendorf, Erin M. Byrt, Anna B. Owczarczyk, Minori Ohashi, Steven R. SchneblyAbstract:AbstractSucrose, raffinose and Stachyose accumulate as stored soluble carbohydrates in embryos during soybean [Glycine max L. (Merrill)] seed development and maturation. Raffinose and Stachyose in soybean feed products are not digested by humans, chickens or pigs, resulting in flatulence and reduced nutritional value. Soybean lines selected for low raffinose and low Stachyose (LRS) or low raffinose, low Stachyose and low phytin (LRSP1, LRSP2) concentrations in mature seeds were compared to a CHECK line with normal raffinose, Stachyose and phytin. To determine whether increasing the supply of free cyclitols to immature embryos of these lines results in increased accumulation of galactosyl cyclitols, isolated immature embryos free of maternal tissues were fed solutions containing either d-chiro-inositol, myo-inositol or d-pinitol, or a control solution without cyclitols, for 24 h. Embryos were precociously matured by slow drying for 14 d with daily transfers to stepwise lower relative humidities. Soluble carbohydrates were extracted from axis and cotyledon tissues of mature, dry embryos and analysed by high-resolution gas chromatography. Axis and cotyledons from LRS, LRSP1 and LRSP2 embryos had low concentrations of Stachyose compared to CHECK embryos after feeding a control solution without cyclitols. Feeding d-chiro-inositol to isolated embryos increased fagopyritol B1 accumulation in embryos of all lines. Feeding myo-inositol increased Stachyose accumulation in LRSP1 and LRSP2 cotyledons. Feeding d-pinitol increased free d-pinitol in cotyledons of all lines but increased galactopinitol A and galactopinitol B only in LRS cotyledons. Supplying additional d-chiro-inositol to immature embryos can enhance accumulation of fagopyritol B1 in mature embryos of low-raffinose and low-Stachyose or low-raffinose, low-Stachyose and low-phytin soybeans.
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free cyclitol unloading from seed coats on stem leaf pod explants of low raffinose low Stachyose low phytin soybean
Seed Science Research, 2010Co-Authors: Suzanne M Kosina, Steven R. Schnebly, Ralph L. ObendorfAbstract:Raffinose, Stachyose and phytin are undesirable compounds for soybean food and animal feed products. In seeds, raffinose and Stachyose are believed to contribute to desiccation and cold stress tolerance. Thus, removal of these compounds from soybean by genetic mutation has resulted in a more commercially desirable composition, but potentially less physiologically viable seeds. In an effort to develop a method to improve viability and seed storability in soybean, stem–leaf–pod explants of three low raffinose, low Stachyose lines, two of which were also low in phytin, and a check line were fed solutions containing d - chiro -inositol, myo -inositol or d -pinitol, free cyclitols which unload through the seed coat to the developing embryo where they accumulate as fagopyritols, galactinol and galactopinitols, respectively, during seed maturation. Increased galactopinitol and fagopyritol accumulation may substitute for the roles of raffinose and Stachyose in low raffinose, Stachyose and phytin seeds. Explants of all lines unloaded d - chiro -inositol, myo -inositol and d -pinitol. Fed d - chiro -inositol accumulated in leaf tissues demonstrating uptake into explants. Fed d - chiro -inositol and myo -inositol accumulated in pod wall and seed coat tissues of one or more lines. The results indicate that d - chiro -inositol was unloaded from the seed coat to the embryo in increased amounts after feeding. The potential use of increased maternal d - chiro -inositol for synthesis of fagopyritols in embryos to improve seed performance in low-Stachyose and low-phytin soybean seeds is supported. The seed coat cup unloading of fed free cyclitols may provide a model system to test effective unloading of upregulated maternally synthesized cyclitols.
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accumulation of soluble carbohydrates during seed development and maturation of low raffinose low Stachyose soybean
Crop Science, 2009Co-Authors: Ralph L. Obendorf, Elizabeth M. Sensenig, Suzanne M Kosina, Angela D Zimmerman, Qianyi Zhang, Alexander Castillo, Elizabeth G Bryant, Steven R. SchneblyAbstract:Soybean [Glycine max (L.) Merr.] seeds accumulate sucrose, raffinose family oligosaccharides (RFO), phytin, and small amounts of galactopinitols and fagopyritols during normal seed maturation. RFO and phytin are indigestible by non-ruminant animals and contribute to decreased feed efficiency, reduced mineral adsorption, and phosphorous pollution in manure. Low raffinose, Stachyose, and phytin seed may have imbibitional chilling sensitivity and reduced field emergence. The objective was to characterize the profiles of soluble carbohydrates in cotyledons, axis, and seed coats of low raffinose and Stachyose (LRS) seeds expressing the mutant stc1 phenotype; in low raffinose, Stachyose, and phytin (LRSP1, LRSP2) seeds expressing the mutant mips phenotype; and in normal raffinose, Stachyose, and phytin (CHECK) seeds expressing the Stc1 and Mips phenotype during 17 stages of soybean seed development, maturation, and desiccation. Seventy percent of RFO accumulated after maximum seed dry weight during seed desiccation. LRS, LRSP1, and LRSP2 seeds had low RFO, but LRS seeds accumulated higher galactinol and di- and tri-galactosides of myo-inositol, D-chiro-inositol, and D-pinitol earlier during seed maturation than CHECK, LRSP1, and LRSP2. LRSP1 and LRSP2 seed had low RFO and low galactosyl cyclitols during maturation and were reported to have imbibitional chilling sensitivity and reduced field emergence.
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soluble carbohydrates in mature soybean seed after feeding d chiro inositol myo inositol or d pinitol to stem leaf pod explants of low raffinose low Stachyose lines
Plant Science, 2008Co-Authors: Ralph L. Obendorf, Elizabeth M. Sensenig, Minori Ohashi, Timothy E Osullivan, Suzanne M Kosina, Steven R. SchneblyAbstract:Abstract Soybean [Glycine max L. (Merrill)] embryos accumulate sucrose, raffinose and Stachyose during seed development and maturation. Raffinose and Stachyose in soybean feed products are not digested by monogastric animals, resulting in flatulence and fewer nutrients for growth. Three lines of soybean selected for low-raffinose and low-Stachyose (LRS) or low-raffinose, low-Stachyose and low-phytin (LRSP1, LRSP2) concentrations in mature seed were compared to a CHECK line with normal concentrations of raffinose, Stachyose and phytin. To determine if increasing the supply of free cyclitols to developing seed of these lines results in increased accumulation of galactosyl cyclitols, soybean stem-leaf-pod explants of each line were fed solutions containing 10 mM d -chiro-inositol, 10 mM myo-inositol, or 10 mM d -pinitol, or a control solution without cyclitols for 7 days. Explants were air-dried for 14 days. Soluble carbohydrates were extracted from axis, cotyledon, and seed coat tissues of mature, dry seed and analyzed by high-resolution gas chromatography. Axis and cotyledons from mature seed of all modified lines had low concentrations of Stachyose compared to the CHECK line when fed a control solution without cyclitols. Feeding d -chiro-inositol to explants increased fagopyritol B1 accumulation in mature seed from all lines. Feeding myo-inositol to explants increased Stachyose accumulation in mature seed of LRSP1, but feeding d -pinitol to explants did not alter soluble carbohydrate composition of mature seed. These responses demonstrate that an increase in the supply of free d -chiro-inositol in maternal tissues can result in enhanced accumulation of fagopyritols in mature seed of low-raffinose and low-Stachyose or low-raffinose, low-Stachyose and low-phytin soybean.
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soluble saccharides and cyclitols in alfalfa medicago sativa l somatic embryos leaflets and mature seeds
Plant Science, 1995Co-Authors: Marcin Horbowicz, Bryan D. Mckersie, Ralph L. Obendorf, D R ViandsAbstract:Abstract Soluble carbohydrates were identified and quantified during development, maturation and desiccation of somatic embryos of alfalfa ( Medicago sativa L.) and compared to soluble carbohydrates in leaflets and mature seeds, to relate changes in soluble carbohydrates to maturation events. Somatic embryos have elevated levels of sucrose. However, in contrast to mature seeds, alfalfa somatic embryos do not accumulate d -pinitol or the galactosyl derivatives of d -pinitol such as galactopinitol A, galactopinitol B and ciceritol. Lower levels of Stachyose accumulate during maturation of somatic embryos, but Stachyose increases to levels in mature seeds during desiccation of somatic embryos. When Stachyose accumulation is limited in somatic embryos, galactinol and digalactosyl myo -inositol increase. Reducing sugars decline to low levels during desiccation of somatic embryos and sucrose: oligosaccharide ratio decreases from 2.7 to 0.9, approaching the ratio 0.2 to 0.3 in mature dry seeds. Except for the lack of pinitol and galactosyl pinitols, changes in soluble carbohydrates during the maturation and desiccation of alfalfa somatic embryos are typical of changes occurring in mature seeds that have been reported to be associated with desiccation tolerance and storability.
Steven R. Schnebly - One of the best experts on this subject based on the ideXlab platform.
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Cyclitol galactosides in low-raffinose, low-Stachyose soybean embryos after feeding d-chiro-inositol, myo-inositol or d-pinitol
Seed Science Research, 2013Co-Authors: Elizabeth M. Sensenig, Ralph L. Obendorf, Erin M. Byrt, Anna B. Owczarczyk, Minori Ohashi, Steven R. SchneblyAbstract:AbstractSucrose, raffinose and Stachyose accumulate as stored soluble carbohydrates in embryos during soybean [Glycine max L. (Merrill)] seed development and maturation. Raffinose and Stachyose in soybean feed products are not digested by humans, chickens or pigs, resulting in flatulence and reduced nutritional value. Soybean lines selected for low raffinose and low Stachyose (LRS) or low raffinose, low Stachyose and low phytin (LRSP1, LRSP2) concentrations in mature seeds were compared to a CHECK line with normal raffinose, Stachyose and phytin. To determine whether increasing the supply of free cyclitols to immature embryos of these lines results in increased accumulation of galactosyl cyclitols, isolated immature embryos free of maternal tissues were fed solutions containing either d-chiro-inositol, myo-inositol or d-pinitol, or a control solution without cyclitols, for 24 h. Embryos were precociously matured by slow drying for 14 d with daily transfers to stepwise lower relative humidities. Soluble carbohydrates were extracted from axis and cotyledon tissues of mature, dry embryos and analysed by high-resolution gas chromatography. Axis and cotyledons from LRS, LRSP1 and LRSP2 embryos had low concentrations of Stachyose compared to CHECK embryos after feeding a control solution without cyclitols. Feeding d-chiro-inositol to isolated embryos increased fagopyritol B1 accumulation in embryos of all lines. Feeding myo-inositol increased Stachyose accumulation in LRSP1 and LRSP2 cotyledons. Feeding d-pinitol increased free d-pinitol in cotyledons of all lines but increased galactopinitol A and galactopinitol B only in LRS cotyledons. Supplying additional d-chiro-inositol to immature embryos can enhance accumulation of fagopyritol B1 in mature embryos of low-raffinose and low-Stachyose or low-raffinose, low-Stachyose and low-phytin soybeans.
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free cyclitol unloading from seed coats on stem leaf pod explants of low raffinose low Stachyose low phytin soybean
Seed Science Research, 2010Co-Authors: Suzanne M Kosina, Steven R. Schnebly, Ralph L. ObendorfAbstract:Raffinose, Stachyose and phytin are undesirable compounds for soybean food and animal feed products. In seeds, raffinose and Stachyose are believed to contribute to desiccation and cold stress tolerance. Thus, removal of these compounds from soybean by genetic mutation has resulted in a more commercially desirable composition, but potentially less physiologically viable seeds. In an effort to develop a method to improve viability and seed storability in soybean, stem–leaf–pod explants of three low raffinose, low Stachyose lines, two of which were also low in phytin, and a check line were fed solutions containing d - chiro -inositol, myo -inositol or d -pinitol, free cyclitols which unload through the seed coat to the developing embryo where they accumulate as fagopyritols, galactinol and galactopinitols, respectively, during seed maturation. Increased galactopinitol and fagopyritol accumulation may substitute for the roles of raffinose and Stachyose in low raffinose, Stachyose and phytin seeds. Explants of all lines unloaded d - chiro -inositol, myo -inositol and d -pinitol. Fed d - chiro -inositol accumulated in leaf tissues demonstrating uptake into explants. Fed d - chiro -inositol and myo -inositol accumulated in pod wall and seed coat tissues of one or more lines. The results indicate that d - chiro -inositol was unloaded from the seed coat to the embryo in increased amounts after feeding. The potential use of increased maternal d - chiro -inositol for synthesis of fagopyritols in embryos to improve seed performance in low-Stachyose and low-phytin soybean seeds is supported. The seed coat cup unloading of fed free cyclitols may provide a model system to test effective unloading of upregulated maternally synthesized cyclitols.
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accumulation of soluble carbohydrates during seed development and maturation of low raffinose low Stachyose soybean
Crop Science, 2009Co-Authors: Ralph L. Obendorf, Elizabeth M. Sensenig, Suzanne M Kosina, Angela D Zimmerman, Qianyi Zhang, Alexander Castillo, Elizabeth G Bryant, Steven R. SchneblyAbstract:Soybean [Glycine max (L.) Merr.] seeds accumulate sucrose, raffinose family oligosaccharides (RFO), phytin, and small amounts of galactopinitols and fagopyritols during normal seed maturation. RFO and phytin are indigestible by non-ruminant animals and contribute to decreased feed efficiency, reduced mineral adsorption, and phosphorous pollution in manure. Low raffinose, Stachyose, and phytin seed may have imbibitional chilling sensitivity and reduced field emergence. The objective was to characterize the profiles of soluble carbohydrates in cotyledons, axis, and seed coats of low raffinose and Stachyose (LRS) seeds expressing the mutant stc1 phenotype; in low raffinose, Stachyose, and phytin (LRSP1, LRSP2) seeds expressing the mutant mips phenotype; and in normal raffinose, Stachyose, and phytin (CHECK) seeds expressing the Stc1 and Mips phenotype during 17 stages of soybean seed development, maturation, and desiccation. Seventy percent of RFO accumulated after maximum seed dry weight during seed desiccation. LRS, LRSP1, and LRSP2 seeds had low RFO, but LRS seeds accumulated higher galactinol and di- and tri-galactosides of myo-inositol, D-chiro-inositol, and D-pinitol earlier during seed maturation than CHECK, LRSP1, and LRSP2. LRSP1 and LRSP2 seed had low RFO and low galactosyl cyclitols during maturation and were reported to have imbibitional chilling sensitivity and reduced field emergence.
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soluble carbohydrates in mature soybean seed after feeding d chiro inositol myo inositol or d pinitol to stem leaf pod explants of low raffinose low Stachyose lines
Plant Science, 2008Co-Authors: Ralph L. Obendorf, Elizabeth M. Sensenig, Minori Ohashi, Timothy E Osullivan, Suzanne M Kosina, Steven R. SchneblyAbstract:Abstract Soybean [Glycine max L. (Merrill)] embryos accumulate sucrose, raffinose and Stachyose during seed development and maturation. Raffinose and Stachyose in soybean feed products are not digested by monogastric animals, resulting in flatulence and fewer nutrients for growth. Three lines of soybean selected for low-raffinose and low-Stachyose (LRS) or low-raffinose, low-Stachyose and low-phytin (LRSP1, LRSP2) concentrations in mature seed were compared to a CHECK line with normal concentrations of raffinose, Stachyose and phytin. To determine if increasing the supply of free cyclitols to developing seed of these lines results in increased accumulation of galactosyl cyclitols, soybean stem-leaf-pod explants of each line were fed solutions containing 10 mM d -chiro-inositol, 10 mM myo-inositol, or 10 mM d -pinitol, or a control solution without cyclitols for 7 days. Explants were air-dried for 14 days. Soluble carbohydrates were extracted from axis, cotyledon, and seed coat tissues of mature, dry seed and analyzed by high-resolution gas chromatography. Axis and cotyledons from mature seed of all modified lines had low concentrations of Stachyose compared to the CHECK line when fed a control solution without cyclitols. Feeding d -chiro-inositol to explants increased fagopyritol B1 accumulation in mature seed from all lines. Feeding myo-inositol to explants increased Stachyose accumulation in mature seed of LRSP1, but feeding d -pinitol to explants did not alter soluble carbohydrate composition of mature seed. These responses demonstrate that an increase in the supply of free d -chiro-inositol in maternal tissues can result in enhanced accumulation of fagopyritols in mature seed of low-raffinose and low-Stachyose or low-raffinose, low-Stachyose and low-phytin soybean.
Peter R. Gibson - One of the best experts on this subject based on the ideXlab platform.
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Measurement of Short-Chain Carbohydrates in Common Australian Vegetables and Fruits by High-Performance Liquid Chromatography (HPLC)
Journal of agricultural and food chemistry, 2009Co-Authors: Jane G. Muir, Kelly Liels, Jacqueline Susanne Barrett, Rosemary Rose, Susan Joy Shepherd, Ourania Rosella, Peter R. GibsonAbstract:Fermentable oligo-, di-, and monosaccharides and polyols (FODMAPs) are short-chain carbohydrates that can be poorly absorbed by the small intestine and may have a wide range of effects on gastrointestinal processes. FODMAPs include lactose, fructose in excess of glucose, fructans and fructooligosaccharides (FOS, nystose, kestose), galactooligosaccharides (GOS, raffinose, Stachyose), and sugar polyols (sorbitol, mannitol). This paper describes an analytical approach based on HPLC with ELSD that quantifies the major FODMAPs in 45 vegetables and 41 fruits. Sorbitol and/or mannitol were measured in 18 vegetables (range = 0.09-2.96 g/100 g of fw), raffinose and/or Stachyose in 7 vegetables (0.08-0.68 g/100 g of fw), and nystose and/or kestose in 19 vegetables (0.02-0.71 g/100 g of fw). Apple, pear, mango, clingstone peach, and watermelon all contained fructose in excess of glucose. Sorbitol was measured in 15 fruits (0.53-5.99 g/100 g of fw), mannitol was found in 2 fruits, and nystose or kestose was measured in 8 fruits. Understanding the importance of dietary FODMAPs will be greatly assisted by comprehensive food composition data.
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measurement of short chain carbohydrates in common australian vegetables and fruits by high performance liquid chromatography hplc
Journal of Agricultural and Food Chemistry, 2009Co-Authors: Jane G. Muir, Kelly Liels, Jacqueline Susanne Barrett, Rosemary Rose, Susan Joy Shepherd, Ourania Rosella, Peter R. GibsonAbstract:Fermentable oligo-, di-, and monosaccharides and polyols (FODMAPs) are short-chain carbohydrates that can be poorly absorbed by the small intestine and may have a wide range of effects on gastrointestinal processes. FODMAPs include lactose, fructose in excess of glucose, fructans and fructooligosaccharides (FOS, nystose, kestose), galactooligosaccharides (GOS, raffinose, Stachyose), and sugar polyols (sorbitol, mannitol). This paper describes an analytical approach based on HPLC with ELSD that quantifies the major FODMAPs in 45 vegetables and 41 fruits. Sorbitol and/or mannitol were measured in 18 vegetables (range = 0.09−2.96 g/100 g of fw), raffinose and/or Stachyose in 7 vegetables (0.08−0.68 g/100 g of fw), and nystose and/or kestose in 19 vegetables (0.02−0.71 g/100 g of fw). Apple, pear, mango, clingstone peach, and watermelon all contained fructose in excess of glucose. Sorbitol was measured in 15 fruits (0.53−5.99 g/100 g of fw), mannitol was found in 2 fruits, and nystose or kestose was measured ...
Pollyanna Amaral Viana - One of the best experts on this subject based on the ideXlab platform.
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debaryomyces hansenii ufv 1 intracellular α galactosidase characterization and comparative studies with the extracellular enzyme
Journal of Agricultural and Food Chemistry, 2009Co-Authors: Pollyanna Amaral Viana, Sebastiao Tavares De Rezende, Flavia Maria Lopes Passos, Marcelo P Bemquerer, Jamil S Oliveira, Kadima N Teixeira, Alexandre Martins Costa Santos, Jose C Rosa, Marcelo M Santoro, Valeria Monteze GuimaraesAbstract:Debaryomyces hansenii cells cultivated on galactose produced extracellular and intracellular alpha-galactosidases, which showed 54.5 and 54.8 kDa molecular mass (MALDI-TOF), 60 and 61 kDa (SDS-PAGE) and 5.15 and 4.15 pI values, respectively. The extracellular and intracellular deglycosylated forms presented 36 and 40 kDa molecular mass, with 40 and 34% carbohydrate content, respectively. The N-terminal sequences of the alpha-galactosidases were identical. Intracellular alpha-galactosidase showed smaller thermostability when compared to the extracellular enzyme. D. hansenii UFV-1 extracellular alpha-galactosidase presented higher kcat than the intracellular enzyme (7.16 vs 3.29 s-1, respectively) for the p-nitrophenyl-alpha-D-galactopyranoside substrate. The Km for hydrolysis of pNPalphaGal, melibiose, Stachyose, and raffinose were 0.32, 2.12, 10.8, and 32.8 mM, respectively. The intracellular enzyme was a competitively inhibited by galactose (Ki = 0.70 mM), and it was inactivated by Cu(II) and Ag(I). Enzyme incubation with soy milk for 6 h at 55 degrees C reduced Stachyose and raffinose amounts by 100 and 73%, respectively.
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hydrolysis of oligosaccharides in soybean products by debaryomyces hansenii ufv 1 α galactosidases
Food Chemistry, 2007Co-Authors: Pollyanna Amaral Viana, Sebastiao Tavares De Rezende, Maurilio Alves Moreira, Daniel Luciano Falkoski, Thiago De Almeida Leite, Ines Charnel Jose, Valeria Monteze GuimaraesAbstract:α-Galactosides are abundant sugars found in legumes such as soybean. Since humans and monogastric animals lack α-galactosidase in the digestive tract, they are unable to digest these sugars, which induce flatulence. The use of α-galactosidases is promising as a means to overcome this problem, and to increase the consumption of soy products. Immobilized α-galactosidase, derived from Debaryomyces hansenii UFV-1, exhibited an activity of 40 U per g of silica and an activity yield of 50%. The optimum pH of free and immobilized α-galactosidase was 5.0 and the optima temperatures were 60 and 80 °C, respectively. The soymilk Stachyose was completely hydrolyzed by different enzyme forms after incubation for 4 h at 60 °C, while raffinose was reduced by 100%, 25% and 68% by free, immobilized enzymes and permeabilized cells, respectively. The soy molasses treatment with free enzyme for 6 h promoted reduction in Stachyose and raffinose contents by 100% and 50%, respectively.
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extracellular α galactosidase from debaryomyces hansenii ufv 1 and its use in the hydrolysis of raffinose oligosaccharides
Journal of Agricultural and Food Chemistry, 2006Co-Authors: Pollyanna Amaral Viana, Sebastiao Tavares De Rezende, Virginia M Marques, Larissa Mattos Trevizano, Flavia Maria Lopes Passos, Maria Goreti De Almeida Oliveira, Marcelo P Bemquerer, Jamil S Oliveira, Valeria Monteze GuimaraesAbstract:Raffinose oligosaccharides (RO) are the factors primarily responsible for flatulence upon ingestion of soybean-derived products. ROs are hydrolyzed by α-galactosidases that cleave α-1,6-linkages of α-galactoside residues. The objectives of this study were the purification and characterization of extracellular α-galactosidase from Debaryomyces hansenii UFV-1. The enzyme purified by gel filtration and anion exchange chromatographies presented an Mr value of 60 kDa and the N-terminal amino acid sequence YENGLNLVPQMGWN. The Km values for hydrolysis of pNPαGal, melibiose, Stachyose, and raffinose were 0.30, 2.01, 9.66, and 16 mM, respectively. The α-galactosidase presented absolute specificity for galactose in the α-position, hydrolyzing pNPGal, Stachyose, raffinose, melibiose, and polymers. The enzyme was noncompetitively inhibited by galactose (Ki = 2.7 mM) and melibiose (Ki = 1.2 mM). Enzyme treatments of soy milk for 4 h at 60 °C reduced the amounts of Stachyose and raffinose by 100%. Keywords: α-Galactosi...
