• Title/Summary/Keyword: stachyose

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Molecular Characterization of the α-Galactosidase SCO0284 from Streptomyces coelicolor A3(2), a Family 27 Glycosyl Hydrolase

  • Temuujin, Uyangaa;Park, Jae Seon;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.26 no.9
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    • pp.1650-1656
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    • 2016
  • The SCO0284 gene of Streptomyces coelicolor A3(2) is predicted to encode an α-galactosidase (680 amino acids) belonging to glycoside hydrolase family 27. In this study, the SCO0284 coding region was cloned and overexpressed in Streptomyces lividans TK24. The mature form of SCO0284 (641 amino acids, 68 kDa) was purified from culture broth by gel filtration chromatography, with 83.3-fold purification and a yield of 11.2%. Purified SCO0284 showed strong activity against p-nitrophenyl-α-D-galactopyranoside, melibiose, raffinose, and stachyose, and no activity toward lactose, agar (galactan), and neoagarooligosaccharides, indicating that it is an α-galactosidase. Optimal enzyme activity was observed at 40℃ and pH 7.0. The addition of metal ions or EDTA did not affect the enzyme activity, indicating that no metal cofactor is required. The kinetic parameters Vmax and Km for p-nitrophenyl-α-D-galactopyranoside were 1.6 mg/ml (0.0053 M) and 71.4 U/mg, respectively. Thin-layer chromatography and mass spectrometry analysis of the hydrolyzed products of melibiose, raffinose, and stachyose showed perfect matches with the masses of the sodium adducts of the hydrolyzed products, galactose (M+Na, 203), melibiose (M+Na, 365), and raffinose (M+Na, 527), respectively, indicating that it specifically cleaves the α-1,6-glycosidic bond of the substrate, releasing the terminal D-galactose.

Changes of Physicochemical and Microbiological Properties of Gamma-Irradiated Miscellaneous Cereals (감마선 조사한 잡곡류의 물리화학적, 미생물적 특성 변화)

  • 손인숙;김미라
    • Journal of the East Asian Society of Dietary Life
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    • v.8 no.4
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    • pp.412-421
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    • 1998
  • Physicochemical and microbiological properties of red beans, soybeans, barleys, and corns irradiated by gamma-ray at 1.2 kGy, 10.1 kGy, or 30.5 kGy were investigated. Moisture content and crude lipid content of the irradiated cereals showed no significant difference from the nonirradiated group. TBA values of corns irradiated at 30.5 kGy increased. Gamma irradiation reduced the numbers of mesophilic and psychrotrophic bacteria in the cereals. Yeasts and molds were reduced below detection levels even at 1.2 kGy dose-irradiation. There was no significant difference in reducing sugars between the irradiated cereals and the nonirradiated ones except corns. Stachyose, raffinose, and sucrose of soy beans and sucrose of barleys increased by 30.5 kGy dose-irradiation. Loss of riboflavin content of the irradiated samples was not observed except red beans irradiated at 30.5 kGy. In Hunter's color, b value of the red bean powder increased but L value of the soybean powder decreased by the Irradiation. L, a, and b values of the barley powder slightly increased under 30.5 kGy dose-irradiation and b value of the corn powder decreased under 10.1 or 30.5 kGy-dose irradiation.

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Characterization of Extracellular $\alpha$-Galactosidase Produced by Bacillus licheniformis YB-42. ($\alpha$-Galactosidase를 생산하는 Bacillus lichennformis YB-42의 분리와 효소 특성)

  • 김현숙;이경섭;소재호;이미성;최준호;윤기홍
    • Microbiology and Biotechnology Letters
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    • v.32 no.2
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    • pp.128-134
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    • 2004
  • A bacterium producing the $\alpha$-galactosidase was isolated from Korean soybean paste. The isolate YB-42 has been identified as Bacillus licheniformis on the basis on its 16S rRNA sequence, morphology and biochemical properties. The $\alpha$-galactosidase activity was detected in both the culture supernatant and the cell extract of B. licheniformis YB-42. The partially purified extracellular $\alpha$-galactosidase was obtained from the culture supernatant by DEAE-Sepharose column and Q-Sepharose column chromatography. The enzyme showed the maximum activity for hydrolysis of para-nitrophenyl-$\alpha$-D-galactopyranoside (pNP-$\alpha$Gal) at pH 6.5 and $45^{\circ}C$. It was able to hydrolyze oligomeric substrates such as melibiose, raffmose and stachyose to liberate galactose residue, indicating that the a-galactosidase of B. licheniformis YB-42 hydrolyzed $\alpha$-1,6 linkage. The hydrolyzing activity of $\alpha$-galactosidase for both pNP-$\alpha$Gal and melibiose was dramatically decreased by galactose. Both glucose and mannose inhibited the activity for pNP-$\alpha$Gal less than galactose.

Mannanolytic Enzyme Activity of Paenibacillus woosongensis (Paenibacillus woosongensis의 만난분해 효소활성)

  • Yoon, Ki-Hong
    • Korean Journal of Microbiology
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    • v.46 no.4
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    • pp.397-400
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    • 2010
  • The activities of mannanase, ${\beta}$-mannosidase, and ${\alpha}$-galactosidase were detected in culture filtrate of Paenibacillus woosongensis showing mannanolytic activity for locust bean gum. Optimal conditions occurred at pH 5.5 and $60^{\circ}C$ for mannanase toward locust bean gum, pH 6.5 and $50^{\circ}C$ for ${\beta}$-mannosidase toward para-nitrophenyl-${\beta}$-D-mannopyranoside, and pH 6.0-6.5 and $50^{\circ}C$ for ${\alpha}$-galactosidase toward para-nitrophenyl-${\alpha}$-D-galactopyranoside in the culture filtrate, respectively. The mannanolytic enzyme of culture filtrate hydrolyzed mannobiose as well as manno-oligosaccharides including mannotriose, mannotetraose, mannopentaose and mannohexaose. It could also hydrolyze ${\alpha}$-1,6 linked galacto-oligosaccharides such as melibiose, raffinose and stachyose to liberate galactose residue. From these results, it is assumed that P. woosongensis produces three enzymes required for the complete decomposition of galactomannan.

Changes in Isoflavone and Some Characteristics of Chokong of Germinated Soybeans during Pickling in Vinegar (발아콩을 이용한 초콩의 제조 중 Isoflavone 및 특성 변화)

  • Eom, Kwon-Yong;Kim, Joo-Sook;Choi, Hee-Sook;Cha, Bo-Sook;Kim, Woo-Jung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.3
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    • pp.359-365
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    • 2006
  • Changes in isoflavone and oligosaccharides, and some physicochemical properties of Chokong, a pickled and dried soybeans, were investigated during pickling in brewed vinegar at $20^{\circ}C$. The used soybeans were 24 hr germinated soybeans, which was maximumly increased in isoflavone content during germination. The isoflavone contents were significantly increased by approx. 80% in both glycosides and aglycone type of isoflavone after 20 day of pickling at $20^{\circ}C$. The isoflavone values of germinated Chokong were significantly higher than those of ungerminated ones. Pickling the soybeans in vinegar resulted in a rapid initial decrease in oligosaccharides, Particularly in raffinose and stachyose. The pH and soluble solids contents in vinegar increased markedly and L values decreased during initial pickling of 24 hr.

Efficacy of Enzyme Treatment for the Quality Improvement of Soymilk (두유(豆乳)의 품질향상을 위한 효소제(酵素劑) 처리의 효과)

  • Yoo, Jeong-Seon;Lee, Su-Rae
    • Korean Journal of Food Science and Technology
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    • v.20 no.3
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    • pp.426-432
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    • 1988
  • This study was undertaken to investigate for the possible use of enzymes with ${\alpha}-galactosidase$ and protease activities to remove flatulence factors as well as to improve the yield and protein digestibility in soymilk preparation. The volume and protein yield were not increased significantly by enzyme treatment. The solids yield increased by raising treatment pH 6 to 10, the temperature $30^{\circ}C\;to\;60^{\circ}C$ Enzyme treatment brought about a remarkable increase in TCA-soluble nitrogen compounds and a decrease in the contents of flatulence factors raffinose and stachyose.

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Influence of Gibberellic Acid on α-D-Galactosidase Activity in the Grape Berry

  • Kang, Han-Chul;Lee, Seon-Hwa;Kim, Jong-Bum
    • Journal of Applied Biological Chemistry
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    • v.44 no.2
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    • pp.53-58
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    • 2001
  • Glycosidase activities in the grape flesh (Marguerite) were assayed, and the order of activity was marked as follows: ${\alpha}$-D-galactosidase>${\alpha}$-D-mannosidase>${\alpha}$-D-glucosidase>${\beta}$-D-galactosidase>${\beta}$-D-glucosidase. Of these glycosidases, ${\alpha}$- and ${\beta}$-D-galactosidases were prominently expressed by the treatment of gibberellic acid, resulting in 56 and 238% increase of activity, respectively. Most of ${\alpha}$-D-galactosidase was found in the flesh texture, and the activity increase by gibberellic acid occurred mostly in this tissue. The increase in ${\alpha}$-D-galactosidase activity was dependent on the concentration of gibberellic acid treated, showing a positive correlation. Gibberellic acid affected the content of total protein in the grape flesh, 49% increase by 75 ppm treatment. Above this concentration, higher gibberellic acid level did not influence the protein expression. Specific activity of the ${\alpha}$-D-galactosidase still increased, showing 24% increase in activity. Grape flesh subjected by gibberellic acid (100 ppm) resulted in the increased activity against a natural substrate, stachyose, showing 55% increase in activity from the grapes treated with 100 ppm of gibberellic acid. Other natural substrates, such as melibiose and raffinose, were also considerably hydrolyzed, and the extent was similar to that of stachyose hydrolysis. During postharvest storage, ${\alpha}$-D-galactosidase activity in the grape flesh increased by 51% after 20 days and then declined slowly.

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Kinetic Properties of $\alpha$-Galactosidase from Aspergillus niger ATCC 16513 and Soybean(Glycine max. L) (Aspergillus niger ATCC 16513과 대두(Glycine max. L) $\alpha$-galactosidase의 kinetic 성질)

  • Geum, Jong-Hwa;Lee, Jong-Su;Sin, Cheol-Seung
    • The Journal of Natural Sciences
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    • v.5 no.1
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    • pp.53-57
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    • 1992
  • This experiment was carried out to elucidate some kinetic properties of the $\alpha$-galactosidase which produced and purified from Aspergillus niger ATCC 16513 and soybean(Glycine max. L). The Km value of Asp. niger and soybean $\alpha$-galactosidase were 37.0mM and 50.0mM for raffinose and55.5mM and 55.5mM for stachyose, respectively. The activity of Asp. niger and soybean $\alpha$-galactosidase were inhibited by galactose. Among the amino acids in active sites of both Asp. niger and soybean $\alpha$-galactosidase, histidine was identified by chemical modification of diethyl pyrocarbonate. Number of amino acids residues per mole of Asp. niger and soybean $\alpha$-galactosidase were 902 and 286, respectively.

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Changes in Isoflavone and Oligosaccharides of Soybeans during Germination (콩의 발아 중 이소플라본과 올리고당의 변화)

  • Kim, Joo-Sook;Kim, Jong-Goon;Kim, Woo-Jung
    • Korean Journal of Food Science and Technology
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    • v.36 no.2
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    • pp.294-298
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    • 2004
  • Three Korean soybean varieties, Dawon, Taekwang, and Myeongiju-namul were investigated for changes in isoflavone and oligosaccharides contents, and dry weight during germination. Soybeans were soaked for 10 hr in water, followed by 8 days germination at $20^{\circ}C$ under dark condition, Highest isoflavone content measured was Myeongju-namul (1.228 mg/g), followed by Taekwang (0.671 mg/g) and Dawon (0.661 mg/g). Total isoflavone content generally increased during initial germination and decreased thereafter. Maximal increase in isoflavone was 20-50%, particularly in aglycone type such as daidzein and genistein. Raffinose and stachyose contents decreased rapidly during germination, while that of sucrose showed relatively slow decrease. Dry weights of soybeans steadly decreased.

Effects of fermented soybean meal supplementation on the growth performance in sows and piglets

  • Seok Han, Ra;Hyoung Churl, Bae;Myoung Soo, Nam
    • Korean Journal of Agricultural Science
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    • v.48 no.4
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    • pp.807-814
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    • 2021
  • This study sought to evaluate the effects of fermented soybean meal (FSBM) prepared by inoculating Bacillus coagulans NRR1207 and a Kefir starter on sows and Holstein cow's. FSBM has high nutritional value due to the hydrolysis of anti-nutritional factors such as trypsin inhibitors, hemagglutinin, raffinose and stachyose. In particular, it is widely used as a type of livestock feed due to its high protein content. The composition of FSBM is as follows: crude protein 55.15%, crude fat 2.12% and 0.2% KOH solubility 83.17%, it was higher than soybean meal (SBM). In particular, anti-nutritional factors such as trypsin inhibitor, raffinose and stachyose of FSBM were significantly reduced compared to the SBM. The number of lactic acid bacteria, including B. coagulans NRR1207, is 8.63 × 107 CFU·g-1, yeast is 1.1 × 106 CFU·g-1. Offspring numbers, the initial sucking number, sucking days, and weaned numbers of sows fed with FSBM all showed higher values compared to the control group. The average body weight and backfat thickness of sows fed with FSBM increased than those fed with SBM. The weight body of piglets fed with FSBM increased by 1.4 kg compared to the control group. The feed conversion ratio of piglets fed with FSBM was reduced by 10.69% compared to the control group. The results of this study indicate that FSBM can provide beneficial effects with regard to the feeding characteristics of sows and piglets.