• Title/Summary/Keyword: dextranase

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Optimization of an Extracellular Dextranase Production from Lipomyces starkeyi KCTC 17343 and Analysis of Its Dextran Hydrolysates (Lipomyces starkeyi KCTC 17343에 의한 extracellular dextranase 최적생산과 덱스트란 hydrolysates 분석)

  • Chang, Yoon-Hyuck;Yeom, Joong-Hyun;Jung, Kyung-Hwan;Chang, Byung-Chul;Shin, Jung-Hee;Yoo, Sun-Kyun
    • Journal of Life Science
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    • v.19 no.4
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    • pp.457-461
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    • 2009
  • We optimized dextranase culture conditions by batch fermentation using Lipomyces starkeyi KCTC 17343. Furthermore, dextranase was purified by an ultra-membrane, and then dextran hydrolyzates were characterized. Cell growth and dextranase production varied depending on the initial culture pH and temperature. The conditions of optimal dextranase production were met in a pH range of 4-5 and temperature between $25-30^{\circ}C$. At optimal fermentation conditions, total enzyme activity and specific enzyme activity were about 4.85 IU/ml and 0.79 IU/g cells, respectively. The specific growth rate was examined to be $0.076\;hr^{-1}$. The production of dextranase in culture broth was very stably maintained after mid-log phase of growth. The enzyme hydrolyzed dextran into DP (degree of polymerization) 2 to 8 oligodextran series. Analysis of the composition of hydrolysates suggested that the enzyme produced is an endo-dextranase.

Effect of dextranase and glucose-oxidase on the formation of plaque by Streptococcus mutans (Streptococcus mutans의 Plaque 형성에 미치는 Dextranase와 Glucose-oxidase 의 영향)

  • 김윤석;안재현;정광례;이기붕
    • Korean Journal of Microbiology
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    • v.27 no.4
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    • pp.430-435
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    • 1989
  • Dextranase and glucose-osidase was investigated as an anti-plaque agent and a component of dentifrice. In vitro synthesis of the water-insoluble glucan was decreased with increasing amount of dextranase and glucose-oxidase. Dextranase was effective on the decrease of viable S. mutans, and the formation of plaque decreased. But it is not effective on the degradatio of plaque. As a research for addition of enzyme to the dentifrice components, we formulated the Model Dentifrice for stabilization of enzyme. At the Model Dentifrice, we confirmed the stability of enzyme by evalution of activity for a long time.

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Isolation and Identification of Dextranase Production Strains and Enzyme Production (Dextranase 생산균주의 분리, 동정 및 효소생산)

  • Lee, Jong-Tae;Yi, Dong-Heui;Kwak, Yi-Seong;Kim, Young-Ho;Sung, Hyun-Soon;Kim, Chan-Jo
    • Microbiology and Biotechnology Letters
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    • v.23 no.4
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    • pp.405-410
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    • 1995
  • In order to screen dextranase with high dextranolytic activity from microbial origins, dextranase producing fungal isolates were isolated from soil of the Taeion area. 197 strains with dextranolytic activities were isolated, out of which 3 strains with high dextranolytic activities were selected in the first screening. A strain (GR-98) with a best dextranolytic activity was selected in the second screening. The strain was identified to be similiar Aspergillus ustus by the morphological and cultural characteristics. The optimum culture temperature and initial pH for the dextranase production of the strain was 30$\circ$C and 7.0, respectively. The optimum culture medium was composed of 2% dextran, 0.3% KNO$_{3}$, 0.05% K$_{2}$HPO$_{4}$, 0.02% MgSO$_{4}$-7H$_{2}$O, 0.05% KC1, and 2.5 $\mu$g/ml pyridoxamine, and the enzyme production was maximum when the strain was subcultured at 30$\circ$C for 7 days.

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The Effect of Dextranase-Containing Mouthwash in Human Experimental Gingivitis (실험적 치은염에서 dextranase 함유 구강 세정액의 양치 효과)

  • Son, Eun-Ju;Kim, Young-Jun;Kim, Do-Man;Chung, Hyun-Ju
    • Journal of Periodontal and Implant Science
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    • v.31 no.2
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    • pp.401-420
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    • 2001
  • A novel glucanhydrolase from a mutant of Lipomyces starkeyi(KSM 22)has been shown effective in hydrolysis of mutan, reduction of mutan formation by Streptococcus mutans and removal pre-formed sucrose-dependent adherent microbial film and Lipomyces starkeyi KSM 22 dextranase has been strongly bound to hydroxyapatitie. These in vitro properties of Lipomyces starkeyi KSM 22 dextranase are desirable for its application as a dental plaque control agent. This study was performed to determine oral hygiene benefits and safety of dextranase(Lipomyces starkeyi KSM 22 dextranase)-containing mouthwash in human experimental gingivitis. This 3-week clinical trial was placebo-controlled double-blind design evaluating 1U/ml dextranase mouthwash and 0.12% chlorhexidine mouthwash. A total 39 systemically healthy subjects, who had moderate levels of plaque and gingivitis were included. At baseline, 1, 2 and 3 weeks, subjects were scored for plaque(Silness and $L{\ddot{o}e$ plaque index and plaque severity index), gingivitis($L{\ddot{o}e$ and Silness gingival index), and at baseline and 3 weeks of experiment, subjects were scored for plaque(Turesky-Quingley-Hein's plaque index and plaque severity index), tooth stain(Area and severity index system by Lang et al). Additionally, oral mucosal examinations were performed and subjects questioned for adverse symptoms. Two weeks after pre-experiment examinations and a professional prophylaxis, the subjects provided with allocated mousewash and instructed to use 20-ml volumes for 30s twice dailywithout toothbrushing. All the groups showed significant increase in plaque accumulation since 1 week of experiment. During 3 weeks' period, the dextranase group showed the least increase in plaque accumulation of Silness and $L{\ddot{o}e$ plaque index, compared to the chlorhexidine and placebo groups, but chlorhexidine group showed the least increase inplaque accumulation of Turesky-Quingley-Hein's plaque index. As for gingival inflammation, all the groups showed significant increase during 3 weeks of experiment. The dextranase group also showed the least increase in gingival index score, compared to the chlorhexidine as well as the placebo groups. Whereas the tooth stain was increased significantly in the chlorhexidine group, compared to the baseline score and the placebo group since 3 weeks of mouthrinsing. It was significantly increased after 3 weeks in the dextranase group, still less severe than the chlorhexidine group. As for the oral side effect, the dextranase group showed less tongue accumulation, bad taste, compared to the chlorhexidine group. From these results, mouthrinsing with Lipomyces starkeyi KSM 22 dextranase was comparable to 0.12% chlorhexidine mouthwashin inhibition of plaque accumulation and gingival inflammation and local side effects were if anything less frequent and less intense than chlorhexidine, in human experimental gingivitis. All data had provided positive evidence for Lipomyces starkeyi KSM 22 dextranase as an antiplaque agent and suggested that further development of dextranase formulations for plaque control are warranted.

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Cloning and expression of Lipomyces starkeyi dextranase-encoding gene in yeasts

  • Kang, Hee-Kyoung;Park, Ji-Young;An, Joon-Seob;Kim, Seung-Heuk;Kim, Do-Man
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.402-406
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    • 2005
  • Lipomyces starkeyi produces a novel glucanhydrolase containing endo-dextranase and ${\alpha}-amylase$ activities. A cDNA from L. starkeyi encoding a dextranase was isolated and characterized. The 2,052 kb cDNA fragment (lsd1) carrying dextranase gene showed one open reading frame (ORF) composed of 1,824 bp flanked by a 41 bp 5'-UTR and a 184 bp 3'-UTR including a poly(A) tail of 27 bp. The ORF encodes for a 608 amino acid with a predicted molecular mass of 67.6 kDa. There was 77% deduced amino acid sequence identity between the LSD1 dextranase and the dextranase from Penicillium minioluteum. The primary structure of the dextranase from L. starkeyi has distant similarity with enzymes belonging to glycosyl hydrolase family 49. The lsd1 protein was expressed in the Saccharmyces cerevisiae under control of GAL1 promoter and active dextranase was produced.

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Characterization of Exo-dextranase from Aspergillus ustus (Aspergillus ustus의 Exo-dextranase의 특성에 관한 연구)

  • Lee, Kon-Joo;Lee, Hyung-Hoan
    • The Korean Journal of Mycology
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    • v.11 no.1
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    • pp.15-21
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    • 1983
  • Exodextranase from Aspergillus ustus was purified by chromatography and characterized by various conditions. The optimal pH of the purified dextranase was 6.5 and this enzyme was maximally activated at $40^{\circ}C$. The enzyme was stable at the temperature below $50^{\circ}C$. The enzyme was markedly inactivated by $Hg^{2+},\;Cu^{2+},\;KCN\;and\;Co^{2+},\;but\;Ba^{2+},\;Fe^{2+},$ cysteine, EDTA, and ascorbic acid enhanced the activity of the enzyme. The main products from the hydrolysis of dextran incubated with the dextranase were glucose, isomaltotriose and oligosaccharide. When dextran was incubated with the mixture of pullulanase and ${\alpha}-amylase$, it was hydrolyzed into glucose, isomaltose and oligosaccharide. Polysaccharides in the decade teeth powder were hydrolyzed by the dextranase into glucose, isomaltotriose and oligosaccharides. In the hydrolysis of the teeth powder with the mixture of dextranase, pullulanase and ${\alpha}-amylase$, were proved to be similar to the dextran hydrolysates.

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Rational Introduction of Disulfide Bond to Enhance Optimal Temperature of Lipomyces starkeyi $\alpha$-Dextranase Expressed in Pichia pastoris

  • Chen, Lin;Yu, Chao;Zhou, Xiangshan;Zhang, Yuanxing
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1506-1513
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    • 2009
  • $\alpha$-Dextranase, which can hydrolyze dextran, is largely used in the sugar industry. However, a thermostable $\alpha$-dextranase is needed to alleviate the viscosity of syrups and clean blocked machines. Thus, to improve the optimal temperature of Lipomyces starkeyi $\alpha$-dextranase expressed by Pichia pastoris, the rational introduction of a de novo designed disulfide bond was investigated. Based on the known structure of Penicillium minioluteum dextranase, L. starkeyi $\alpha$-dextranase was constructed using homology modeling. Four amino acids residues were then selected for site-directed mutagenesis to cysteine. When compared with the wild-type dextranase, the mutant DexM2 (D279C/S289C) showed a more than $13^{\circ}C$ improvement on its optimal temperature. DexM2 and DexM12 (T245C/N248C, D279C/S289C) also showed a better thermal stability than the wild-type dextranase. After the introduction of two disulfide bonds, the specific activity of DexM12 was evaluated and found to be two times higher than that of the wild-type. Moreover, DexM12 also showed the highest $V_{max}$.

Properties of Carbohydrase Prepared from Lipomyces starkeyi JLC26 (Lipomyces starkeyi JLC26에서 유래된 Carbohydrase의 특성)

  • Jun, Sun-Mee;Kim, Do-Man;Kim, Do-Won
    • KSBB Journal
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    • v.14 no.6
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    • pp.713-717
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    • 1999
  • We have isolated a dextranase and amylase constitutive and hyper-producing mutant, Lipomyces starkeyi JLC26, from Lipomyces starkeyi ATCC74054 after mutation using UV irradiation. After partial purification of dextranase and amylase (together DXAMase;both activities were always co-purified) by ammonium sulfate precipitation, CM-Sepharose column chromatography, the specific activities of amylase and dextranase were 5367 and 3045 unit/mg, respectively. The pH effects for activity and stabiligy of both enzymes were similar to each other: Optimum pH and temperature for activity sere at 5.5 and 37$^{\circ}C$ and optimum ranges for stability were at pH 2.5-5.5 and 4-55$^{\circ}C$, respectively. The reaction end products of dextranase and amylase activities were found to the typical for those of endo-dextranase and endo-amylase. When the carbohydrase and maltotriose were reacted, glucose, maltose, isomaltose, maltotriose, panose and ${\alpha}(1{\rightarrow}6)$glucosylmaltotriose were produced by disproportionation reaction.

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Cloning of a Gene Encoding Dextranase from Lipomyces starkeyi and its Expression in Pichia pastoris

  • Kang, Hee-Kyoung;Park, Ji-Young;Ahn, Joon-Seob;Kim, Seung-Heuk;Kim, Do-Man
    • Journal of Microbiology and Biotechnology
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    • v.19 no.2
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    • pp.172-177
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    • 2009
  • A gene(lsd1) encoding dextranase from Lipomyces starkeyi KSM22 has been previously cloned, sequenced, and expressed in Saccharomyces cerevisiae. The gene consisting of 1,824 base pairs and encoding a protein of 608 amino acids was then cloned into and secretively expressed in Pichia pastoris under the control of the AOX1 promoter. The dextranase productivity of the P. pastoris transformant(pPIC9K-LSD1, 134,000 U/I) was approximately 4.2-fold higher than that of the S. cerevisiae transformant(pYLSD1, 32,000 U/I) cultured in an 8-1 fermentor. Over 0.63 g/l of active dextranase was secreted into the medium after methanol induction. The dextranase of the P. pastoris transformant, as analyzed by SDS-PAGE and Western blotting, showed only one homogeneous band. This dextranase of the P. pastoris transformant showed a broad band near 73 kDa. Rabbit monoclonal antibodies against a synthetic LSD1 peptide mix also recognized approximately 73 kDa.

Penircillium속에 의한 Dextranase 생산에 관한 연구

  • 김옥희;민태익;한문희
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1975.07a
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    • pp.111.1-111
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    • 1975
  • Dextranase는 dextran의 $\alpha-1,6-glycoside$ 결합을 끊는 효소로써 치석을 형성하는데 요인이 되는 dextran을 분해 제거하는 역할을 한다. 본 실험에서는 이러한 dextranase 생산 곰팡이류 중에서 Penicillium 속에 속하는 균주를 선정하여 이 효소 생산의 최적조건을 실험 검토하였다.

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