• Title/Summary/Keyword: dextransucrase

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Enzymatic Synthesis of Puerarin Glucosides Using Leuconostoc Dextransucrase

  • Ko, Jin-A;Ryu, Young Bae;Park, Tae-Soon;Jeong, Hyung Jae;Kim, Jang-Hoon;Park, Su-Jin;Kim, Joong-Su;Kim, Doman;Kim, Young-Min;Lee, Woo Song
    • Journal of Microbiology and Biotechnology
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    • v.22 no.9
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    • pp.1224-1229
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    • 2012
  • Puerarin (P), an isoflavone derived from kudzu roots, has strong biological activities, but its bioavailability is often limited by its low water solubility. To increase its solubility, P was glucosylated by three dextransucrases from Leuconostoc or Streptococcus species. Leuconostoc lactis EG001 dextransucrase exhibited the highest productivity of puerarin glucosides (P-Gs) among the three tested enzymes, and it primarily produced two P-Gs with a 53% yield. Their structures were identified as ${\alpha}$-$_D$-glucosyl-($1{\rightarrow}6$)-P (P-G) by using LC-MS or $^1H$- or $^{13}C$-NMR spectroscopies and ${\alpha}$-$_D$-isomaltosyl-($1{\rightarrow}6$)-P (P-IG2) by using specific enzymatic hydrolysis, and their solubilities were 15- and 202-fold higher than that of P, respectively. P-G and P-IG2 are easily applicable in the food and pharmaceutical industries as alternative functional materials.

Large Increase in Leuconostoc citreum KM20 Dextransucrase Activity Achieved by Changing the Strain/Inducer Combination in an E. coli Expression System

  • Ko, Jin-A;Jeong, Hyung-Jae;Ryu, Young-Bae;Park, Su-Jin;Wee, Young-Jung;Kim, Do-Man;Kim, Young-Min;Lee, Woo-Song
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.510-515
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    • 2012
  • A recombinant putative dextransucrase (DexT) was produced from Leuconostoc citreum KM20 as a 160 kDa protein, but its productivity was very low (264 U/l). For optimization, we examined enzyme activity in 7 Escherichia coli strains with inducer molecules such as lactose or IPTG. E. coli BL21-CodonPlus(DE3)-RIL exhibited the highest enzyme activity with lactose. Finally, DexT activity was remarkably increased by 12-fold under the optimized culture conditions of a cell density to start induction ($OD_{600}$) of 0.95, a lactose concentration of 7.5 mM, and an induction temperature of $17^{\circ}C$. These results may effectively apply to the heterologous expression of other large DexT genes.

Structural Characteristics of Novel Branched Oligosaccharides Syntesized by a Maltose Acceptor Reaction with Dextransucrase from Leuconostoc mesenteroides M-12 (Leuconostoc mesenteroides M-12 덱스트란수크라제의 말토스 억셉터 반응으로 합성된 새로운 분지 올리고당의 구조 특성)

  • 서현창
    • The Korean Journal of Food And Nutrition
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    • v.10 no.1
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    • pp.102-109
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    • 1997
  • The structures of novel branched oligosaccharides synthesized by the acceptor reaction with dextransucrase from Leuconostoc mesenteriodes M-12 were proposed in accordance with the results obtained from enzymatic hydrolyses and a partial acid hydrolysis. The structure of branched oligosaccharide B4 was shown to be 62-O-$\alpha$-D-kojibiosylmaltose. Branched oligosaccharide B5 was shown to be 63-O-$\alpha$-D-kojibiosylpanose. By reacting the acceptor reaction products with endodextranase a novel branched oligosaccharide (D4) could be produced. D4 was derived from the result of endodextranase hydrolysis of oligosaccarides synthesized by the second acceptor reaction with dextransucrase and was resistant to endodextranase and glucoamylase. The proposed structure of D4 was 62-O-$\alpha$-D-kojibiosylisomaltose. Formation pattern of the acceptor reaction products smaller than d.p. 6 with linear or branched linkage was also shown.

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Effect of Reaction Time on the Rheological Properties of Dextran Formed Solution Produced by Crude Dextransucrase from Leuconostoc rnesenteroides Sikhae (Leuconostoc mesenteroides Sikhae의 Crude Dextransucrase 반응시간에 따른 Dextran 생성액의 유변성)

  • 박춘상;이철호
    • Microbiology and Biotechnology Letters
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    • v.20 no.3
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    • pp.316-323
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    • 1992
  • Studies on the changes in rheological properties, molecular weight distribution and dextran yield after being reacted in lO%(w/w) sucrose concentration were performed with crude dextransucrase produced from Leuconostoc mesenteroides isolated from Sikhae. The reaction rate of dextran formation was monitored by sugar analysis with HPLC and by the changes in apparent viscosity. According to the periodate oxidation test, the dextran produced in this experiment was estimated to have 89% $\alpha$-(1->6) main linkages and 11% $\alpha$-(1->) side linkages. The rheological properties of the dextran solution formed changed with reaction time, and it was related to the changes in molecular weight distribution of dextran as determined by GPC analysis. As the reaction proceeded, the rheological behavior changed from Newtonian to non-Newtonian, showing Binghampseudoplastic and thixothropic flow behavior. The apparent viscosity of dextran formed solution increased with increasing reaction time, reached a maximum value of 2680 cP ($\gamma$=$33.75s^{-1}$, $25^{\circ}C$) by enzyme reaction for 8 hours, and then decreased. The temperature dependency of dextran formed solutions was well expressed by the Arrhenius equation and the activation energy reached a maximum value of 1.69 kcal/mole by enzyme reaction for 8 hours.

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Low Cariogenicity of Maltosyl-erythritol, Major Transglycosylation Product of Erythritol, by Bacillus stearothermophilus Maltogenic Amylase

  • Jeon, Eun-Joo;Jung, Il-Hun;Cho, Kil-Soon;Seo, Eun-Sung;Kim, Do-Man;Lee, Sung-Joon;Park, Kwan-Hwa;Moon, Tae-Wha
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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    • pp.815-818
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    • 2003
  • Maltosyl(G2)-erythritol, produced by the transglycosylation reaction of erythritol with maltotriose by Bacillus stearothermophilus maltogenic amylase, was not utilized either as a substrate for lactic acid production or for water-insoluble glucan synthesis. An inhibition assay of dextransucrase and mutansucrase showed that the dental caries suppression effect of G2-erythritol was greater than that of erythritol.

Synbiotic Synthesis of Oligosaccharides During Milk Fermentation by Addition of Leuconostoc Starter and Sugars

  • Seo, Dong-Mi;Kim, So-Young;Eom, Hyun-Ju;Han, Nam-Soo
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1758-1764
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    • 2007
  • Synthesis of oligosaccharides during milk fermentation was attempted by inoculating Leuconostoc citreum with Lactobacillus casei, Lb. delbrueckii subsp. bulgaricus, and Streptococcus thermophilus as starters. Dextransucrase of Ln. citreum worked as a catalyst for the transglycosylation reaction of sugars; sucrose was added as the glucose donor, and lactose or maltose acted as the acceptor compound for the reaction. When 4% sucrose was added in milk, glucosyl-lactose was synthesized (about 1%, w/v) after 1-2 days of fermentation at 15 or $25^{\circ}C$. Alternatively, when sucrose and maltose (2% each, w/v) were added, panose (about 1 %, w/v) and other isomaltooligosaccharides were made in a day at $15-35^{\circ}C$. Growth patterns of lactobacilli and streptococci starters were not affected by the coculture of leuconostoc starter, but the rate of acid synthesis was slightly slowed at every temperature. Addition of sugars in milk did not give any adverse effect on the lactate fermentation. Accordingly, the use of leuconostoc starter and addition of sugars in milk allowed the production of oligosaccharides-containing fermented milk, and application of this method will facilitate the extensive development of synbiotic lactate foods.

Quantitative Analysis of Leuconostoc mesenteroides and Lactobacillus plantarum Populations by a Competitive Polymerase Chain Reaction

  • Koh, Young-Ho;Kim, Myoung-Dong;Han, Nam-Soo;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.12 no.5
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    • pp.801-806
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    • 2002
  • A multiplex competitive polymerase chain reaction (PCR) method was developed for the rapid identification and quantification of Leuconostoc mesnteroides and Lactobacillus plantarum populations which are the key microorganisms in kimchi fermentation. The strain-specific primers were designed to selectively amplify the target genes encoding 165 rRNA of L. plantarum and dextransucrase of L. mesenteroides. There was a linear relationship between the band intensity of PCR products and the number of colony forming units of each model organism. The PCR quantification method was compared with a traditional plate-counting method f3r the enumeration of the two lactic acid bacteria in a mixed suspension culture and also applied to a real food system, namely, watery kimchi. The population dynamics of the two model organisms in the mixed culture were reliably predictable by the competitive PCR analysis.

Study on the Improvement of Quality in Jeung-pyun Prepared with Lactic Bacteria Having High Dextransucrase Activity as Starters (Dextransucrase 고활성 젖산균을 이용한 스타터 증편의 품질개선에 관한 연구)

  • Lee, A-Young;Park, Ju-Yeon;Hahn, Young-Sook
    • Korean Journal of Food Science and Technology
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    • v.38 no.3
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    • pp.400-407
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    • 2006
  • Twenty six strains of lactic acid bacteria were isolated from Jeung-pyun batter, among which Tetragenococcus halophilus 1-12 showed highest dextransucrase activity at 36.95 DSU/mg, followed by T. halophilus 5-8 (36.87 DSU/mg protein), T. halophilus 2-12 (32.66 DSU/mg protein), T. halophilus 3-3 (31.43 DSU/mg protein), T. halophilus 3-1 (30.73 DSU/mg protein), T. halophilus 5-12 (29.43 DSU/mg protein), and Leuconostoc mesenteroides subsp mesenteroides 2-9 (28.5 DSU/mg protein). L. mesenteroides subsp mesenteroides 2-9, T. halophilus 1-12, and L. mesenteroides subsp dextranicum 5-13, were selected as starters (0.1, 0.5, and 1.0%) for Jeung-pyun manufacturing. Specific volume of Jeung-pyun added with 1.0% L. mesenteroides subsp dextranicum 5-13 was highest at 2.00, and 1.0% T. halophilus 1-12-added Jeung-pyun was lowest at 033. Cross-sectional observation of Jeung-pyun showed Jeung-pyun added with 0.5% L. mesenteroides subsp mesenteroides 2-9 was uniformly formed. Number of air holes of Jeung-pyun increased with increasing amount of added T. halophilus 1-12. Increasing addition of L. mesenteroides subsp dextranicum 5-13 resulted in more uniform air holes and volumes. Hardness, gumminess, and chewiness of Jeung-pyun added with 0.5 and 2.0% L. mesenteroides subsp dextranicum 5-13 decreased significantly in compasion to control groups. Jeung-pyun added with 0.5% L. mesenteroides subsp dextranicum 5-13 showed excelIent overall sensory desirability ranking of 8.500.

Production and Characterization of New Structured-Oligosaccharides from Immobilized Mixed-enzyme Reactor (고정화 혼합효소를 이용한 새로운 구조의 올리고당 생산 및 특성 연구)

  • ;;;;;Seiya Chiba;Atsuo Kimura
    • KSBB Journal
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    • v.15 no.1
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    • pp.88-95
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    • 2000
  • We have produced new-structured oligosaccharides using immoobilized mixed-enzyme reactor of destransucrase from Leuconostoc mesenteroides B-512FMCM and $\alpha$-amylase from Aspergillus oryzae. The reactors of immobilized mixed-exzyme beads were more efficient for the production of oligosaccharides than that of each immobilized enzyme bead in stirred-tank reactior(STR) or in packed-bed reactor(PCR). In continuous flow reactor, the immobilized mixed-enzyme bead in PBR was more stable than in STR, and 52% of initial yield was maintained for 200 hr. New structured-oligosaccharides (NOS) reduced the change of pH in the culture of Streptococcus mutans. It also showed an inhibitory effect on the growth of Staphylococcus aureus.

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Characterization of Streptococcus mutans isolated from Human Dental Plaque 2. Streptococcal Polysaccharide. (충치에서 분리한 Streptococcus mutans에 관하여 2)

  • 이건주;이배함
    • Korean Journal of Microbiology
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    • v.18 no.4
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    • pp.180-187
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    • 1980
  • Cariogenic Strptococcus mutans produces a constitutive extracellular enzyme dextransucrase or glucosyltransferase that is capable of hydrolying sucrose and synthesizing the glucose polymer dextran. In this work we investigated to the dextrans produced by eight strains of Strptococcus mutans. After, 30hours the synthesized polysaccharide is 1.86mg to 4.41mg per ml on sucrose medium, and the polysaccharide is similar. Polysaccharide syntheiezd by enzyme in cell free medium is 11.4 mgto 2.36mg per ml after 10 hours.

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