• Title/Summary/Keyword: raffinose

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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 Chemical Components in Persimmon Leaves (Diospyros kaki Thunberg) during Growth (감잎의 성장시기에 따른 화학성분 변화)

  • 김종국;이원영
    • Journal of the East Asian Society of Dietary Life
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    • v.12 no.1
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    • pp.32-37
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    • 2002
  • In order to promote the utilization of persimmon leaf as food, chemical components in freeze dried Persimmon leaves were analyzed. The proximate composition was composed of moisture(79.65%), crude protein(17.97%), crude fat(1.33%), ash(2.37%), crude fiber(2.01%). During growth, moisture content was decreased and crude fat, crude fiber and ash were increased, respectively. Free sugar was composed of glucose, fructose, sucrose, raffinose. Nonvolatile organic acid contents were composed of oxalic acid, levulenic acid, magic acid and citric acid in the order. 18 amino acids of total amino acid in persimmon leaves were detected and major amino acid were glutamic acid, aspartic acid, leucine, phenylalanine, arginine and valine. The soluble tannin and vitamin C of persimmon leaves were 6859.37 mg% and 1487.12 mg%. During growth, its contents increased and then decreased after 20th June.

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Production and Characterization of α-Galactosidases from Two Bacillus licheniformis Isolates (Bacillus licheniformis 분리균 2종의 α-Galactosidase 생산성과 효소특성)

  • Jin, Hyun Kyung;Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.43 no.3
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    • pp.195-203
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    • 2015
  • Two bacterial strains, Bacillus licheniformis YB-1413 and YB-1414, producing extracellular α-galactosidase, were obtained from homemade Doenjang. On the basis of their biochemical properties, 16S rRNA sequences and random amplified polymorphic DNA patterns by polymerase chain reaction, they were found to be somewhat different from one another. α-Galactosidase productivities of the two isolates were increased by wheat bran, but drastically decreased by melibiose, raffinose and sucrose which were used as carbon sources. The enzyme productivities were increased by yeast extract as a nitrogen source with maximum levels of 1.87 U/ml for YB-1413 and 1.69 U/ml for YB-1414, respectively. The enzymes of both isolates exhibited maximum activity for hydrolysis of para-nitrophenyl-α-D-galactopyranoside (pNP-αGal) under reaction conditions of pH 6.0 and 45℃. Their hydrolyzing activities for pNP-αGal were drastically decreased by the addition of low concentrations of ribose and galactose. They were capable of hydrolyzing completely α-1,6 linked galactosyl residue in melibiose, raffinose and stachyose, which are known to be anti-nutritional factors in products of soybean and legume. In relation to the latter, the isolates YB-1413 and YB-1414 have potential applicability in improving soybean-fermented foods and the nutritional value of soybean feed.

Isolation and Identification of Korean type Streptococcus mutans (한국형 Streptococcus mutans의 분리 및 동정)

  • 현성희;장성렬;최영길
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.250-258
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    • 1989
  • S. mutans known as a causative causative agent of dental caries was isolated from a carious lesion of Korean in the present study. The physiological, biochemical characteristics and polysaccharide pattern of these isolates were compated with those of four laboratory strains ; AHT(a), FA-1F(b), LM7(e), and OMZ65(g). One hundred strains of oral streptococci were isolated from dental caries sites of Korean (one male and one female). Among these, 3 strains were identified as S. mutans. These strains were able to grow on selective media MS, MST, MSP, MSP1, MSB, MSBT and were stained dark pink when sprayed with solutions of mannitol and TTC. So, these strains were called strain 108, 110, and 120, respectively. Strain 108, 110, and 120 were bacitracin resistnt. As these strains contained particularly hippurate hydrolysis enzyme, they were distinguished from laboratory strains. Apart from laboratory strains, the strain 108 was not capable of fermenting lactose, the strain 110 was not able to ferment sorbitol, inulin, melibiose, raffinose and the strain120 was incapable of fermenting inulin, raffinose. All fractions of extracellular and ecll bound polysaccharide of the strain108, 110, and 120 were consisted of more glucan than fructan. Aside from laboratory strains, the isolated strains were composed of more water-insoluble glucans related adherence on solid surface than water-soluble. According to these results, the strain108, 110, and 120 had native characteristecs of S. mutans, but they were different from laboratory strains in some characteristics. Therefore, each of them was given a name to S. mutans KHC108, KHC110, and KHC120, respcetively.

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Purification and properties of soybean ${\alpha}-galactosidase$ (대두 ${\alpha}-galactosidase$의 정제 및 성질)

  • Keum, Jong-Hwa;Oh, Man-Jin;Kim, Seong-Yeol
    • Applied Biological Chemistry
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    • v.34 no.3
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    • pp.249-257
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    • 1991
  • To elucidate enzymatic properties of ${\alpha}-galactosidase$ (EC 3, 2, 1, 22) from germinated soybean, changes in the enzyme activities and oligosaccharide contents during germination of soybean were determined. ${\alpha}-Galactosidase$ from germinated soybean was purified by ammonium sulfate fractionation, ion exchange chromatography and gel filtration. Their chemical and enzymatic properties was investigated. ${\alpha}-galactosidase$ activity of sobeam was maximized when it was germinated at $25^{\circ}C$ for 120 hour. Raffinose and stachyose in soybean were decomposed completely after 96 hours and 120 hours of germination, respectively. Soybean ${\alpha}-galactosidase$ was purified by 6.6 fold by ammonium sulfate fractionation, ion exchange chromatography on DEAE-Cellulose and Sephadex A-50, and gel filtration on Sephadex G-150. Its specific activity was 825 Units/mg protein and the yield was 2.5% of the total activity of crude extracts. The purified ${\alpha}-galactosidase$ of soybean was found to be homogeneous by polyacrylamide gel electrophoresis and by HPLC. Isoelectric point of soybean ${\alpha}-galactosidase$ was determined analytical isoelectric focusing to be pH 4.8. The soybean ${\alpha}-galactosidase$ was monomeric and its molecular weight was estimated to be 30,000 by SDS-PAGE. The optimal temperature and pH for the soybeam ${\alpha}-galactosidase$ activity were $40^{\circ}C$ and pH 6.0 and 75% of its activity was lost by heating at $60^{\circ}C$ for 10 min. The enzyme was appeared to have higher affinity to raffinose than to stachyose. The Km value of soybean enzyme was 5.3 mM for ${\rho}-nitrophenyl-{\alpha}-D-galactopyranoside$ and the activation energy on PNPG was calculated to be 13.02 Kcal per mole.

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Changes in Oligosaccharides and Sensory Quality of Soymilk During Germination (대두발아에 의한 콩우유의 과당과 관능적품질의 변화)

  • Kim, Woo-Jung;Yoon, Suk-Kwon;Lee, Chun-Young
    • Korean Journal of Food Science and Technology
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    • v.18 no.5
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    • pp.382-387
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    • 1986
  • The effect of germination of soybeans on chemical and sensory qualities of soymilk were investigated. The soybeans were soaked and germinated at $25^{\circ}C$ for 5 days prior to grinding. The result showed that a rapid initial decrease in the contents of raffinose and stachyose and a slight increase in protein yield were found after 2 days of germination. Undesirable flavor such as beany and grassy was minimum and total acceptability was maximum for the soymilk prepared from soybeans germinated for two days. Changes in roasted nutty odor and taste were almost identical to the changes of total acceptability during-five day germination.

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Quantitative Analysis of Free Amino Acids and Free Sugars in Steamed and Roasted Green Tea by HPLC (HPLC에 의한 증제와 볶음 녹차 중의 유리 아미노산과 유리당의 정량)

  • Ko, Young-Su;Lee, In-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.3
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    • pp.301-304
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    • 1985
  • Changes of free amino acids and free sugars in steamed and roasted green tea were determined after heat treatment at $110^{\circ}C$. Sixteen kinds of free amino acids and four kinds of free sugar were analyzed by HPLC. Free amino acids isolated were aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, histidine, lysine and arginine. Free sugars were sucrose, glucose, fructose, and raffinose. After the heat treatment, free amino acids and free sugars were decreased considerably.

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Biochemical characterization of Bacillus thuringiensis, 23 serovars (Biochemical thuringiensis, 23 serovars의 생화학적 특성)

  • Lee, Hyung-Hoan;Park, Mi-Yeoun;Lee, Chang-Woon
    • Microbiology and Biotechnology Letters
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    • v.14 no.2
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    • pp.205-208
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    • 1986
  • The 23 serovars of Bacillus thuringiensis strain were commonly gram-positive and motile, formed endotoxin crystals, produced acid and alkali in the KIA media, and acid from glucose, hydrolyzed starch, and reduced nitrate but did not produce H$_2$S, oxidase and indole, did not decompose lysine, ornithine, phenylalanine, malonate, lactose, dulcitol, adonitol, inositol, sorbitol, arabinose, raffinose, rhamnose, maltose, and xylose. Eighteen serovars were positive in the MR tests and 15 in the VP tests. Four serovars used citrate. Five serovars produced urease, 5 $CO_2$ from glucose, 2 DNase, and 15 lecithinase. Twelve serovars decomposed arginine, 11 did sucrose, 2 manitol, and 9 salicin Serovar tohokuensis did not hemolyze, but the others did.

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Characterization of Extracellular \alpha-galactosidase Produced by Streptomyces sp. YB-4. (균체외 \alpha-galactosidase를 생산하는 Streptomyces sp. YB-4의 분리 및 효소 특성)

  • 김소영;조기행;김창진;박동진;윤기홍
    • Microbiology and Biotechnology Letters
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    • v.30 no.4
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    • pp.332-338
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    • 2002
  • A strain YB-4 producing the extracellular $\alpha$-galactosidase was isolated from soil, and has been identified as Streptomyces sp. on the basis of its cultural, morphological and physiological properties. The partially purified $\alpha$-galactosidase was most active on paranitrophenyl-$\alpha$-D-galactopyranoside at pH 6.0 and 6$0^{\circ}C$. The enzyme retained 90% of its maximum activity between pH 4.0 and pH 10.0 after pre-incubation for 1 h. The enzyme was able to hydrolyze oligomeric substrates such as melibiose, raffinose and stachyose to liberate galactose residue, indicating that the $\alpha$-galactosidase of Steptomyces sp. YB-4 hydrolyzed $\alpha$-1,6 linkage.

Interaction between Lactobacillus acidophilus and Saccharomyces uvarum on Utilization of Galacto-oligosaccharides in Soymilk (Lactobacillus acidophilus와 Saccharomyces uvarum의 혼합배양에 의한 두유의 발효 중 당이용에 미치는 작용)

  • 유주현;진효상;류인덕
    • Microbiology and Biotechnology Letters
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    • v.17 no.6
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    • pp.533-538
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    • 1989
  • The enhanced growth and lactic fermentation of L. acidophilus, when mixed with S. uvarum, was investigated. Spent medium of S. uvarum, at 10%, stimulated the growth and lactic fermentation of L. acidophilus, and also increased the content of monosaccharide while decreased the contents of sucrose, raffinose, and stachyose in soymilk. While single culture of L. acidophilus consumed only the monosaccharides in soymilk, the mixed culture of L. acidophilus and S. uvarum consumed almost all the oligosaccharides as well as the monosaccharides in soymilk. Thus it was assumed that S. uvarum converted the oligosaccharides into monosaccharides so that L. acidophilus can produce more lactic acid and cell mass by using the increased monosaccharides.

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