• Title/Summary/Keyword: glucanase

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(1-3, 1-4)-$\beta$-Glucan and Starch Contents and Their Hydrolytic Enzyme Activities in Developing Barley Kernels (등숙 중인 보리 종실중 (1-3, 1-4)-$\beta$-Glucan과 전분 함량 및 이들의 가수분해효소 활성)

  • 윤성중;박상래;유남희
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.4
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    • pp.403-409
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    • 1997
  • To obtain information on the accumulation of (1-3, 1-4)-$\beta$-glucans during kernel maturation, (1-3, 1-4)-$\beta$-glucan contents and (1-3, 1-4)-$\beta$-glucanase activities were determined in developing kernels of the two Korean cooking barley varieties, Neulssalbori and Saessalbori. (1-3, 1-4)-$\beta$-Glucan contents in kernels at 5 and 10 days after anthesis(DAA) were very low and the contents increased rapidly in kernels at 15 to 25 DAA. (1-3, 1-4)-$\beta$-Glucan content in kernels at harvest was about 3.5 to 4% of kernel dry matter. (1-3, 1-4)-$\beta$-Glucanase activities were relatively higher in younger kernels but the levels of the activity were very low compared with those in germinating kernels. A significant negative correlation was observed between (1-3, 1-4)-$\beta$-glucan contents and (1-3, 1-4)-$\beta$-glucanase activities. Low levels of (1-3, 1-4)-$\beta$-glucanase activites in kernels at 15 to 30 DAA, however, may indicate that (1-3, 1-4)-$\beta$-glucanases have little effect on the final content of (1-3, 1-4)-$\beta$-glucans in barley kernels. Starch contents and $\alpha$-amylase activities were also determined in developing barley kernels. Starch contents increased rapidly as kernels matured and the content at harvest was about 60% of kernel dry matter. Relativley higher levels of $\alpha$-amylase activities in kernels at the earlier developmental stage decreased rapidly as kernels matured.

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Characterization of Bacillus licheniformis B1 ${\beta}$-1,4-Glucanase Overproduced in Escherichia coli (대장균에서 과잉생산된 Bacillus licheniformis B1의 ${\beta}$-1,4-Glucanase 특성)

  • Song, Hye-Jung;Kim, Hwang-Yeon;Hwang, Jae-Sung;Kim, Han-Bok
    • Korean Journal of Microbiology
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    • v.46 no.1
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    • pp.68-72
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    • 2010
  • The ${\beta}$-1,4-glucanase gene of Bacillus licheniformis B1 was expressed in Esherichia coli BL21, and a protein with a mass of 50 kDa that was soluble was overproduced. A protein with a mass of 37 kDa was secreted from B. licheniformis. It seems that the ${\beta}$-1,4-glucanase produced in E. coli contained the leader peptide and unprocessed carboxy-terminal region, but its processing occurred in the carboxyterminal in Bacillus. The optimal temperature of ${\beta}$-1,4-glucanase was $40^{\circ}C$. The enzyme still had 76% maximal activity at $60^{\circ}C$. The optimal pH of the enzyme was 7. The enzyme retained considerable activities over the weak-acidic, neutral, and weak-basic pH range. Acidic fungal cellulases are used in food, detergent, pulp, paper, textile industries. However, studies about neutral and alkaline cellulase are not enough. The cellulase developed in this study may be useful for industrial applications in the fields of biofuel development.

Molecular Cloning and Functional Expression of Extracellular Exo-β-(1,3)-Glucanase from Candida fermentati SI (Candida fermentati SI의 exo-β-(1,3)-glucanase유전자의 클로닝 및 그 특성)

  • Lim, Yu-Mi;Kim, Bong-Ki;Kim, Sang-Jun;So, Jai-Hyun;Kim, Won-Chan
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.317-323
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    • 2016
  • An isoflavone glucosidase that catalyzes the hydrolysis of isoflavone glucosides into glucose and corresponding aglycones was purified from Candida fermentati SI. The N-terminal sequence was determined to be GLNCDYCN. We designed degenerate primers on the basis of these amino acid sequences and successfully cloned the full structural gene sequence of the isoflavone glucosidase using inverse PCR. The exo-β-(1,3)-glucanase gene consists of 1227 base-pair nucleotides, encoding a 408-amino-acid sequence that shares 41–96% amino acid homology with other yeast exo-β-(1,3)-glucanases belonging to glycoside hydrolase family 5. The recombinant exo-β-(1,3)-glucanase was expressed in Pichia pastoris X-33, using a pPICZA vector system, and further characterized. The molecular mass of the purified exo-β-(1,3)-glucanase was estimated by SDS-PAGE to be 47 kDa. The optimal pH and temperature were pH 4.5 and 40℃, respectively. The Km values of the purified exo-β-(1,3)-glucanase for daidzin and genistin were 0.12 mM and 0.14 mM, respectively. The Vmax values of the purified isoflavone glucosidase were 945.03 U/mg for daidzin and 835.92 U/mg and for genistin.

Activities of Hydrolytic Enzymes in Barley Malts Prepared under Different Germination Conditions (발아조건별 당화용 엿기름의 분해효소 활성도)

  • Mok, Chul-Kyoon;Lee, Young-Tack
    • Applied Biological Chemistry
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    • v.42 no.4
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    • pp.324-329
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    • 1999
  • Barley malts were prepared at 15, 18 and $21^{\circ}C$ for $3{\sim}6$ days, and assayed for ${\beta}-glucanase$, ${\alpha}-amylase$ and ${\beta}-amylase$ activities. ${\beta}-Glucanase$ activity increased markedly during earley germination and reached maximum at the 6th day of germination. ${\beta}-Glucanase$ activity in six-rowed barley malt was much higher than that in two-rowed malt. ${\beta}-Glucanase$ activity was associated with reduction in ${\beta}-glucan$ content during germination. ${\beta}-amylase$ activity was also considerably higher in two-rowed barley, and increased continuously during 6-day germination. ${\beta}-Amylase$ activity was the lowest at $15^{\circ}C$, the highest at $18^{\circ}C$, and intermediate at $21^{\circ}C$ of germination temperature. Considerable amount of ${\beta}-amylase$ was detected in ungerminated raw barley, and this enzymatic activity tended to increase during 6-day germination. Diastatic power, measure of starch-saccharifying enzyme, in six-rowed malt was $1.4{\sim}1.6$ fold higher than in two-rowed malt. Germination at $18^{\circ}C$ for $5{\sim}6$ days was suggested to be the optimum condition for manufacturing good quality malts, in terms of enhanced starch-degrading enzymatic activity.

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Breeding of Ethanol-producing and Ethanol-tolerant Saccharomyces cerevisiae using Genome Shuffling (Genome shuffling을 이용한 에탄올 생산 및 내성 효모 균주의 육종)

  • Park, A-Hwang;Kim, Yeon-Hee
    • Journal of Life Science
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    • v.23 no.10
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    • pp.1192-1198
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    • 2013
  • To improve yeast strains for bioethanol production, yeasts with ethanol tolerance, thermotolerance, and ${\beta}$-1,3-glucanase activity were bred using yeast genome shuffling. Saccharomyces cerevisiae $BY4742{\Delta}exg1$/pAInu-exgA, which has extracellular ${\beta}$-1,3-glucanase activity, and the Aspergillus oryzae and S. cerevisiae YKY020 strains, which exhibit ethanol tolerance and thermotolerance, were fused by yeast protoplast fusion. Following cell fusion, four candidate cells (No. 3, 9, 11, and 12 strains) showing thermotolerance at $40^{\circ}C$ were selected, and their ethanol tolerance (7% ethanol concentration) and ${\beta}$-1,3-glucanase activity were subsequently analyzed. All the phenotypes of the two parent cells were simultaneously expressed in one (No. 11) of the four candidate cells, and this strain was called BYK-F11. The BYK-F11 fused cell showed enhanced cell growth, ethanol tolerance, ${\beta}$-1,3-glucanase activity, and ethanol productivity compared with the $BY4742{\Delta}exg1$/pAInu-exgA and YKY020 strains. The results prove that a new yeast strain with different characters and the same mating type can be easily bred by protoplast fusion of yeasts.

Using Enzyme Supplemented, Reduced Protein Diets to Decrease Nitrogen and Phosphorus Excretion of White Leghorn Hens

  • Jacob, Jacqueline P.;Ibrahim, Sami;Blair, Robert;Namkung, Hwan;Paik, In Kee
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.12
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    • pp.1743-1749
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    • 2000
  • An experiment was conducted to investigate the effect of supplementation of commercial phytase and ${\beta}-glucanase$ to wheat-soybean meal based layer diets. Control (17% CP) and reduced protein (13.5% CP) diets were compared with and without phytase and/or ${\beta}-glucanase$. Reducing dietary crude protein levels reduced the amount of N excreted by laying hens with no adverse affect on egg production or overall feed conversion ratio. There was, however, a slight reduction in average egg weight. When phytase was added to the control protein diets it was possible to reduce the level of dicalcium phosphate in the diet without a loss in performance and daily P output was reduced significantly. When phytase was added to the reduced protein diets, however, there was a dramatic loss in performance in the last four weeks of the study. Supplementation of ${\beta}-glucanase$ to wheat based layer diet did not appear to have beneficial affects in terms of laying performance and reducing nitrogen or phosphorus excretion. Combination of phytase and ${\beta}-glucanase$ had no positive effects on laying performance or reduction of DM, N and P.

The Probiotic and Adherence Properties of Lactobacillus reuteri Pg4 Expressing the Rumen Microbial β-Glucanase

  • Yu, B.;Liu, J.R.;Hsiao, F.S.;Lee, T.T.;Chiou, P.W.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.9
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    • pp.1324-1329
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    • 2008
  • This study was conducted to evaluate the potential of the transformed Lactobacillus reuteri Pg4 (T-Pg4) harboring the ${\beta}$-glucanase gene as a poultry probiotic. The probiotic properties of the T-Pg4 strain were evaluated in vitro by their adherence capability and acid and bile salt tolerance, and were evaluated in vivo by their survival and adhesion in the gastrointestinal tract (GIT) of specific-pathogen-free (SPF) chickens. The results showed that the T-Pg4 strain exhibited resistance to acidic conditions and contact with bile salt, and adhered efficiently to the crop and intestinal epithelial cells of chickens in vitro. The T-Pg4 strain also could survive and colonize the gastrointestinal epithelium of the experimental SPF chickens in vivo. In addition, radial enzyme diffusion was used to demonstrate that the Lactobacillus spp. randomly isolated from the GIT of the SPF chickens fed T-Pg4 possessed ${\beta}$-glucanase secretion capability. These findings have demonstrated that the transformed L. reuteri Pg4 survives transit through the stomach and intestine, and may secrete ${\beta}$-glucanase in the chicken GIT. Therefore, it is suggested that this organism could be used as a multifunctional poultry probiotic.

Effects of the Addition of \beta-Glucanase to Barley-based Layer Diet (산란계 사료에 \beta-Glucanase 첨가가 보리의 이용성에 미치는 효과)

  • 이정호;이규호;이영철;오상집
    • Korean Journal of Poultry Science
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    • v.21 no.3
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    • pp.195-205
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    • 1994
  • Effects of the addition of \beta-glucanase to barley-based layer diets were examined by feeding 200 Leghorn layers with corn-based (Control) and \beta-glucanase supplemented diets (Barley+ Enzyme). The results obtained are sumrrarized as follows. 1. There were no siginificant (P>0.05) differences in hen-day egg production(%) and average egg weight between two treatments, indicating that the \beta-glucanase supplemented barley could successfully replace the commonly used corn in the layer diets. 2. Although there was no statistical difference (P>0.05) between two treatments, the daily feed consumption was numerically high in layers fed the barly diet compared to the corn-based diet. 3. Availabilities of crude fat and crude fiber of the barley diet were significantly poor (P<0.05) as compared to corn diet. 4. The \beta-glucarase supplementation depressed the viscosity of barley diets and excreta from therm. 5. Both serum and egg yolk cholesterol were not significantly affected by the addition of \beta-glucarase in the barley based diet. Our data indicate that the barley grain supplemented with \beta-glucarase can be sucessfully used as an energy source of layer diet when there is a price advantage. Although some possibilities to produce low cholesterol egg were recognized in this study, further studies pertaining to long-term feeding experiment and elucidaton of the metabolic interrelationship between serum and yolk cholesterol, are required to confirm the result.

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The role and characterization of .betha.-1, 3-glucanase in biocontrol of fusarium solani by pseudomonas stutzeri YPL-1

  • Lim, Ho-Seong;KiM, Sang-Dal
    • Journal of Microbiology
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    • v.33 no.4
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    • pp.295-301
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    • 1995
  • An antifungal Pseudomonas stutzeri YPL-1 produced extracellular chitinase and .betha.-1, 3-glucanase that were key enzymes in the decomposition of fungal hyphal walls. These lytic extracellular enzymes markedly inhibited mycelial growth of the phytopathogenic fungus Fusarium solani. A chitinase from P. stutzeri YPL-1 inhibited fungal mycelial growth by 87%, whereas a .betha.-1, 3-glucanase from the bacterium inhibited growth by 53%. Furthermore, co-operative action of the enzymes synergistically inhibited 95% of the fungal growth. The lytic enzymes caused absnormal swelling and retreating on the fungal hyphal walls in a dual cultures. Scanning electron microscopy clearly showed hyphal degradation of F. solani in the regions interacting with P. stutzeri YPL-1. In an in vivo pot test, P. stutzeri YPL-1 proved to have biocontrol ability as a powerful agent in controlling plant disease. Planting of kidney bean (Phaseolus vulgaris L.) seedlings with the bacterial suspension in F. solani-infested soil significantly suppressed the development of fusarial root-rot. The characteristics of a crude preparation of .betha.-1, 3-glucanase produced from P. stutzeri YPL-1 were investigated. The bacterium detected after 2 hr of incubation. The enzyme had optimum temperature and pH of 40.deg.C and pH 5.5, respectively. The enzyme was stable in the pH range of 4.5 to 7.0 and at temperatures below 40.deg.C, with a half-life of 40 min at 60.deg.C.

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Expression Patterns of Enzymes in Different Tissues of Oil Seed Rape (Brassica napus L.) Seedling (유료용 유채 유식물의 조직내 효소의 발현 패턴)

  • Song, Yong-Su;Seo, Dong-Jun;Lee, Bok-Rye;Jung, Woo-Jin
    • Journal of Applied Biological Chemistry
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    • v.52 no.2
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    • pp.51-57
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    • 2009
  • To investigate expression patterns of chitinase, ${\beta}$-1,3-glucanase and peroxidase involved in biological control of phytopathogens, three oil seed rapes (Capitol, Pollen and Saturnin) were used. Activities of the enzymes in old leaves were $9.7{\sim}11.8$ unit/mg protein in chitinase, $11.1{\sim}17.3$ unit/mg protein in ${\beta}$-1,3-glucanase and $0.6{\sim}1.7$ unit/mg protein in peroxidase. Activities of the enzymes in roots were $39.2{\sim}49.0$ unit/mg protein in chitinase, $49.9{\sim}62.0$ unit/mg protein in ${\beta}$-1,3-glucanase and $2.4{\sim}3.8$ unit/mg protein in peroxidase. Chitinase and ${\beta}$-1,3-glucanase activity were the highest level in Saturnin leaves and in Capitol roots while activities of those were the lowest level in Capitol leaves. Also, chitinase and ${\beta}$-1,3-glucanase and peroxidase activity were the lowest level in Saturnin roots. Active bands of chitinase isoform in leaves (73, 51, 40, 34, and 29 kDa) and in roots (100, 57 34, and 29 kDa) tissues showed in the SDS-PAGE gel. Active bands of ${\beta}$-1,3-glucanase isoform in leaves and roots (75 and 55 kDa) tissues showed on the SDS-PAGE gel. Active staining of peroxidase showed the strongest level in leaves and roots of Pollen. Active bands of peroxidase isoform in leaves (122, 114, and 93 kDa) and in roots (135, 122, 114, and 93 kDa) tissues showed on the Native-PAGE gel. These results indicated that establishment of expression pattern of enzymes in rape tissues could play as an important role with respect to resistance of plant pathogens in rape.