• Title/Summary/Keyword: Insoluble glucan

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Purification and Characterization of Glucosyltransferase and Fructosyltransferase in Leuconostoc mesenteroides NRRL B-1149 (Leuconostoc mesenteroides NRRL B-1149의 Glucosyltransferase와 Fructosyltransferase의 분리와 특성 연구)

  • Lee Jin Ha;Park Jun Seong;Lee Hee Sun;Kim Do Man
    • KSBB Journal
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    • v.19 no.5
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    • pp.368-373
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    • 2004
  • The optimal condition for the production of a glucan and a fructan synthesizing enzymes from Leuconostoc mesenteroides NRRL B-1149 were studied based on the different medium compositions. Response surface methodology was applied to find the optimistic condition showing the relationship between the fermentation response (enzyme activities) and the fermentation variable concentrations of yeast extract, peptone concentration, K2HP04 concentration and sucrose. Optimum medium composition for both enzymes production was $0.75\%$ yeast extract, $0.72\%$ peptone, $1\%$ K2HP04 and $2.17\%$ sucrose. Using this medium, the activities produced in culture was 0.90 U/m~ for glucosyltransferase (GTase) and 0.96 U/ml for fructosyltransferase (FTase). After purification of 1149FTase by consecutive chromatographies using Sephadex G-150 and DEAE-Sepharose, a 1149FTase of 210 kDa on $7\%$ polyacrylamide gel was isolated and it synthesized soluble fructan. The 1149GTase showed a band of 180 kDa on $8\%$ polyacrylamide gel after purification using Bio-Gel P-100 gel chromatography and DEAE-Sepharose ion exchange chromatography and it synthesized insoluble glucan. The linkages of polymers were determined by methylation using Hakomori reagent and following NMR analysis. The glucan was composed of a(1~6) and a(1~3) linkages and the fructan was levan.

Enzymatic Modification of Cellulose Using Leuconostoc mesenteroides B-742CBM Dextransucrase

  • Kim, Do-Man;Kim, Young-Min;Park, Mi-Ran;Ryu, Hwa-Ja;Park, Don-Hee;Robyt, John F.
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.529-533
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    • 1999
  • In addition to catalyzing the synthesis of dextran from sucrose as a primary reaction, dextransucrase also catalyzes the transfer of glucose from sucrose to other carbohydrates that are present or are added to the reaction digest. We have synthesized new glucans having new structures and new characteristics, by transferring D-glucose of sucrose to $\alpha$-cellulose and by using the constitutive dextransucrase obtained from Leuconostoc mesenteroides B-742CBM. The final reaction products were composed of soluble- and insoluble-glucans. The yields of soluble- and insoluble-glucans were theoretically 21% $\pm$ 2.2 and 68% $\pm$ 5.1, respectively. The remainder of the reaction products was recovered as a mixture of olgiosaccharides that could not be precipitated by 67%(v/v) ethanol. Treating the modified glucans with endo-dextranase and/or cellulase, oligosaccharides were produced that were not formed from the hydrolysis of native cellulose or B-742CBM dextran. The modification of the cellulose was confirmed by methylation and acid hydrolysis of the soluble-and insoluble-glucan. Both (1->4) and(1->6) glycosidic linkages were found in both of the glucans.

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Dental caries suppression effect of highly branched and modified oligosaccharides

  • Seo, Eun-Seong;Park, Hyeon-Jeong;Kim, Ji-Hui;Lee, Gi-Yeong;Kim, Do-Man
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.239-242
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    • 2002
  • We have synthesized branched oligosaccharides (BOS) by the mixed-culture fermentation (MBOS), fructosyltransferase (FBOS) or glucosyltransferase (GBOS) with high concentration of sucrose (3M). MBOS was further modified as iron and sulfate-oligosaccharides. The modified MBOS were stable at high temperatures (up to $140^{\circ}C$) and low pHs (2 to 4). Most highly branched and modified oligosaccharide (0.34%, w/v) effectively inhibited fructose release from sucrose by Streptococcus mutans 6715 mutansucrase. FBOS, GBOS, iron-MBOS inhibited the mutansucrase activities from Streptococcus sobrinus about 46.8%, 49.2% and 43.1%, respectively. Most highly branched and modified oligo- saccharides (0.5%, w/v) effectively inhibited the fonnation of insoluble glucan and adherence of S. mutans or S. sobrinus cell in the presence of sucrose. Modified oligosaccharides affected the growth and acid production of oral pathogens. Cytotoxicity test showed that highly branched and modified oligosaccharides was non-toxic.

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Effect of Asarum sieboldii Extracts on the Growth, Acid Production, Adhesion, and Insoluble Glucan Synthesis of Streptococcus mutans (세신 추출물의 Streptococcus mutans에 대한 성장, 산생성, 부착 및 비수용성 글루칸 합성억제에 미치는 영향)

  • Yu Hyeon Hee;Seo Se Jeong;Kim Yeon Hwa;Lee Heung Soo;Kim Kang Ju;Jeon Byung Hun;You Yong Ouk
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.3
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    • pp.666-671
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    • 2003
  • Dental caries are the most commonly occurring chronic diseases in the dental field. Because of increasing sugar consumption and extension of average human life, these diseases are widely found all over the world as the most typical cause for a person to lose a tooth. Therefore, the development of more effective, substantial and safe preventive agents against dental caries is strongly required. Streptococcus mutans is known as the causative bacterial playing the most important role informing plaque and it is being noticed as major causative bacteria of dental caries. The present study was designed to investigate the effect of Asarum sieboldii Miquel(Aristolochiaceae) extracts on the growth, acid production, adhesion, and insoluble glucan synthesis of Streptococcus mutans(S. mutans). Both methanol and aqueous extracts showed concentration dependent inhibitory activity against the growth and acid production of S. mutans, and produced significant inhibition at the concentration of 100, 1,000 and 2,000 μg/ml compared to the control group(p<0.05 - p<0.01). The extracts markedly inhibited S. mutans adherence to HA treated with saliva, and cell adherence was repressed by more than 50% at the concentration of 10 μg/ml and complete inhibition was observed at the concentration of 2,000 μg/ml. On the activity of glucosyltransferase which synthesizes water insoluble glucan from sucrose, methanol and aqueous extracts showed more than 70% inhibition over the concentration of 1,000 μg/ml. Hence, we conclude that Asarum sieboldii might be a candidate of anticaries agent.

Anticaries Effect of Ethanol Extract of Terminalia chebula

  • Lee, Moonkyung;Hwang, Young Sun
    • Journal of dental hygiene science
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    • v.21 no.2
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    • pp.119-126
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    • 2021
  • Background: Dental caries is mainly composed of various cellular components and is deposited around the tooth surface and gums, causing a number of periodontal diseases. Streptococcus mutans is commonly found in the human oral cavity and is a significant contributor to tooth decay. The use of antibacterial ingredients in oral hygiene products has demonstrated usefulness in the management of dental caries. This study investigated the anticaries effect of the ethanol extract of Terminalia chebula (EETC) against S. mutans and their cytotoxicity to gingival epithelial cells. Methods: The EETC was prepared from T. chebula fruit using ethanol extraction. Disk diffusion, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and colony forming unit (CFU) were analyzed to investigate the antimicrobial activity of the EETC. Glucan formation was measured using the filtrate of the bacterial culture medium and sucrose. Gene expression was analyzed using reverse transcription-polymerase chain reaction (RT-PCR). Cytotoxicity was analyzed via the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Results: The antibacterial activity of the EETC was explored using disc diffusion and CFU measurements. The MIC and MBC of the EETC were 10 and 20 ㎍/ml, respectively. EETC treatment decreased insoluble glucan formation by S. mutans enzymes and also resulted in reduced glycosyltransferase B (gtf B), gtf C, gtf D, and fructosyltransferase (ftf), expressions on RT-PCR. In addition, at effective antibacterial concentrations, EETC treatment was not cytotoxic to gingival epithelial cells. Conclusion: These results demonstrate that the EETC is an effective anticaries ingredient with low cytotoxicity to gingival epithelial cells. The EETC may be useful in antibacterial oral hygiene products for the management of dental caries.

Physicochemical Properties and Cooking Quality of Naked Waxy Barley (Saechalssal bon) (새찰쌀보리의 물리화학적 특성 및 취반특성)

  • Jun, Hyun-Il;Cha, Mi-Na;Song, Geun-Seoup;Yoo, Chang-Sung;Kim, Yun-Tae;Kim, Young-Soo
    • Food Science and Preservation
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    • v.18 no.2
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    • pp.165-170
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    • 2011
  • The physicochemical properties and cooking quality of Saechalssal bori (25% pearled ratio), which is a naked waxy barley, were investigated. The amylose, insoluble dietary fiber (IDF), soluble dietary fiber (SDF), total dietary fiber (TDF), and ${\beta}$-glucan contents of Saechalssal bori were 5.2, 10.8, 9.5, 20.3, and 3.7%, respectively. Pasting temperature, peak and final viscosity, and setback of Saechalssal bori were $66.5^{\circ}C$, 383.2, 231.3, and 55.6 RVU, respectively. The water absorption, expansibility, and soluble solid of Saechalssal bori were 232.2, 405.3, and 3.5%, respectively. As the ratio of water to grain increased, L value increased, whereas a and b values were decreased. The sensory evaluation showed that wateriness and overall acceptability increased with increasing ratio of water to grain, resulting in determining 2.1 times as the optimum ratio of water to grain. The cooked Saechalssal bori prepared using optimum condition had hardness (1.2 kg), cohesiveness (4.0), springiness (1.0), gumminess (5.1), chewiness (5.3), adhesiveness (0.2 kg), and A/H (0.8), respectively.

Molecular Characterization of a ${\beta}$-1,4-Endoglucanase Gene from Bacillus subtilis H12

  • Oh, Jin-Hwan;Cha, Jeong-Ah;Yoon, Min-Ho
    • Applied Biological Chemistry
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    • v.51 no.4
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    • pp.299-304
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    • 2008
  • A ${\beta}$-1,4-endoglucanase gene from Bacillus subtilis H12 was cloned into Escherichia coli JM109 (pBC8) and sequenced. The endoglucanase gene with an insert DNA of 2.5 kb possessed an open reading frame of 1,500 bp encoding a mature protein of 499 amino acids with a calculated molecular mass of 55 kDa. The deduced amino acid sequence showed similarity to those of the known neutral cellulase genes of B. subtilis PAP115 (99.2%) and BSE616 (97.8%), as well as the alkaline gene of Bacillus sp. N4 (55.1%). The endoglucanase activity expressed by E. coli (pBC8) was localized in the periplasmic fraction (80%) and the cytoplasmic fraction (20%). An endoglucanase was purified from the periplasmic fraction by performing gel filtration and anion exchange chromatography. The molecular weight of the purified enzyme was estimated to be 31 kDa by SDS-PAGE, and the maximum activity occurred at pH 7 and $40^{\circ}C$. The enzyme easily hydrolyzed soluble substrates such as carboxymethyl cellulose and barely ${\beta}$-glucan, whereas the sigmacell and xylan, the known insoluble substrates, were not entirely hydrolyzed.

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.

A STUDY ABOUT THE INDUCTION OF MUTANASE FROM STREPTOMYCES (Streptomyces의 mutanase 유도에 관한 연구)

  • Yang, Kyu-Ho;Chung, Jin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.23 no.3
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    • pp.764-773
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    • 1996
  • The mutan containing $\alpha$-1,3 bond is an insoluble portion of glucan which is the main component of dental plaque. The secretion of mutanase was assessed with mutan-digesting Streptomyces isolated from soil, and the factors affecting its activity was studied, obtaining the following result. Mutan-digesting Streptomyces was identified as Streptomyces exfoliatus by its characteristics. The effect of dextranase was identified on the media containing blue dextran. A clear zone was produced by Streptomyces exfoliatus on the media containing blue mutan, so showing the secretion of mutanase. A clear zone was significantly produced on the media overlayed with agar containing blue mutan. A clear zone was produced at 2 days after the inoculation of Streptomyces exfoliatus on the media containing below a concentration of 0.025% glucose, at 3 days on the media containing 0.05 % glucose, and at 4 days on the media containing 0.1 % glucose. Mutan-digestion wasn't appeared early by adding other carbohydrates. The higher concentration of peptone, the later appearance of clear zone was on the media containing below a concentration of 0.1 % peptone. These results indicated that the secretion of mutanase was identified from mutan-digesting Streptomyces on the media containing blue mutan, and a clear zone was appeared lately on the media containing higher amount of glucose.

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Study on the development of preventive agent of dental caries from biological active materials Development of disc PAHA for an artificial tooth and preventive effect on dental caries from plant extracts (생물학적 활성물질에서 치아우식 예방제 개발에 관한 연구 I. 인조치아 disc PAHA의 제조 및 식물추출물들의 치아우식 예방효과)

  • 이기용;조효상윤정원허태련
    • KSBB Journal
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    • v.8 no.2
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    • pp.126-132
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    • 1993
  • The objective of this study was to develop an artificial dentin for easy handle and accurate observation of the mechanism on dental caries and to screen biologically active materials from the extracts of traditional plants and fruits for prevention of early dental cares. In order to produce disc PAHA (artificial dentin), the powdered hydroxylapatite was immobilized in a 20% polyacrylamide gel. The characteristics of disc PAHA was very similar to the surface, figure and lattice of human enamel. After decalcification in 0.1M citric acid based on observation with SEM. The critical point of decalcification of disc PAHA by acids was found to be pH 5.0-5.5, which was hi agreement with human enamel. The degree of decalcification from disc PAHA in 0.1M citric acid solution was sixfold higher than that of human enamel. This result suggested that disc PAHA would be useful as a substitute of human enamel for in vitro experiment. The extracts of garlic and Flower Apple A, B seemed to inhibit growth of S. mutans. Especially, when the 300$\mu\ell$ of its extracts added to the medium to incubate S. mutans, F. apple B showed strongly an inhibitory effect in both the growth of S. mutans and the synthesis of insoluble glucan.

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