• Title/Summary/Keyword: ${\beta}$-glucanase

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Comprehensive Analysis of the Expression of Twenty-Seven β-1, 3-Glucanase Genes in Rice (Oryza sativa L.)

  • Hwang, Du Hyeon;Kim, Sun Tae;Kim, Sang Gon;Kang, Kyu Young
    • Molecules and Cells
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    • v.23 no.2
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    • pp.207-214
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    • 2007
  • lant ${\beta}$-1, 3-glucanases are involved in plant defense and in development. Very little data are available on the expression of rice glucanases both in developmental tissues and under various stresses. In this study, we cloned and characterized twenty-seven rice ${\beta}$-1, 3-glucanases (OsGlu) from at total of 71 putative glucanases. The OsGlu genes were obtained by PCR from a cDNA library and were classified into seven groups (Group I to VII) according to their DNA or amino acid sequence homology. Analysis of the expression of the twenty-seven OsGlu genes by Northern blotting revealed that they were differentially expressed in different developmental tissues as well as in response to plant hormones, biotic stress, high salt etc. OsGlu11 and 27 in Group IV were clearly expressed only in stem and leaf and were also induced strongly by SA (5 mM), ABA ($200{\mu}M$), and M. grisea. OsGlu1, 10, 11, and 14 were induced earlier and to higher levels in incompatible M. grisea interaction than in compatible one. Taken together, our findings suggest that the twenty-seven rice OsGlu gene products play diverse roles not only in plant defense but also in hormonal responses and in development.

Antioxidant and α-Glucosidase Inhibitory Effect of Tartary Buckwheat Extract Obtained by the Treatment of Different Solvents and Enzymes (용매 종류와 효소 처리에 따른 쓴 메밀 추출물의 항산화 활성 및 α-Glucosidase 저해 활성의 변화)

  • Kim, Ji-Eun;Joo, Sung-Il;Seo, Ji-Hyun;Lee, Sam-Pin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.8
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    • pp.989-995
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    • 2009
  • Extract yield of tartary buckwheat treated with water, 70% ethanol or methanol were about 13.6%, 7.0% and 6.6%, respectively. Extract yield was greatly increased by the treatment of $\alpha$-amylase indicating 95.1% yield. $RC_{50}$ value of DPPH radical scavenging activity with methanol and 70% ethanol extracts were 34.0 $\mu g$/mL, 40.5 $\mu g$/mL, respectively. The DPPH radical scavenging activity increased when it was treated with $\beta$-glucosidase and cellulase, showing $RC_{50}$ value of 24.7 $\mu g$/mL and 25.0 $\mu g$/mL, respectively. In ABTS radical scavenging activity, methanol extract (100 $\mu g$/mL) showed 30% inhibition. In DPPH or ABTS radical scavenging activities, the treatment of $\beta$-glucanase and $\alpha$-amylase shows the highest and the lowest activities, respectively. In $\alpha$-glucosidase inhibitory effect, 70% ethanol extract showed $RC_{50}$ value of 59.9 $\mu g$/mL, but water extract was not inhibitory effective. The $\alpha$-glucosidase inhibitory effect was the highest in multi enzyme treatment. Content of rutin and quercetin in methanol extract showed higher value with 4400.3 mg% and 71.9 mg%, respectively. The 70% ethanol extract of buckwheat contained rutin of 3459.8 mg% and quercetin of 56.9 mg%. In the treatment of $\beta$-glucanase, the rutin content of ethanol extract increased with 5057.4 mg% and multi-enzyme treatment resulted in the modification of rutin glycoside.

Diversity, Saccharification Capacity, and Toxigenicity Analyses of Fungal Isolates in Nuruk (누룩곰팡이 분리균의 다양성 및 당화능 분석과 독소생산능 조사)

  • Kim, Min Sik;Kim, Sinil;Ha, Byeong-Seok;Park, Hye-Young;BaeK, Seong-Yeol;Yeo, Soo-Hwan;Ro, Hyeon-Su
    • The Korean Journal of Mycology
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    • v.42 no.3
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    • pp.191-200
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    • 2014
  • Nuruk samples collected from various regions in Korea were investigated in terms of fungal contents and diversity. In measurement of colony forming unit (CFU) in Nuruk suspensions on DRBC agar, Nuruk samples MS4, MS8, and MS10 were among the highest fungal density, with $1,278.9{\pm}21.6$ (${\times}10^4$), $1,868.0{\pm}27.7$ (${\times}10^4$), and $775.1{\pm}19.2$ (${\times}10^4$) were among the samples showing the highest fungal density. CFU per 20 mg Nuruk, respectively. The majority of fungal components were yeasts, including Pichia anomala, P. kudriavzevii, Kluyveromyces marxianus, and Saccharomycopsis fibuligera, whereas Aspergillus oryzae and Rhizopus oryzae, the representative Nuruk fungi, were predominant only in the low fungal density Nuruks (MS2, MS5, and MS11). Saccharification capability of the fungal isolates was assessed by measurement of amylase activity in the culture broth. The highest amylase activity was found in A. niger and A. luchuensis, followed by S. fibuligera. A. oryzae and R. oryzae showed fair amylase activity but significantly lower than those of the three fungal species. R. oryzae was suggested to play an additional role in degradation of ${\beta}$-glucan in crop component of Nuruk since R. oryzae was the only fungus that showed ${\beta}$-glucanase activity among the fungal isolates. To confirm the safety of Nuruk, aflatoxigenicity of the isolated Aspergillus was estimated using the DNA markers norB-cypA, aflR, and omtA. All of the isolates turned out to be non-aflatoxigenic as evidenced by the deletion of gene markers, norB-cypA and aflR, and the absence of aflatoxin in the culture supernatants shown by TLC analysis.

Effects of Malt Modification on ${\beta}$-Glucan Solubility and Beer Viscosity (보리의 발아정도가 맥아의 ${\beta}$-glucan 용해성 및 맥주의 점도에 미치는 영향)

  • Lee, Young-Tack
    • Korean Journal of Food Science and Technology
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    • v.40 no.3
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    • pp.360-363
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    • 2008
  • Two barley malt samples were selected at two different stages of germination, a well-modified malt germinated for 96 hr and a poorly-modified malt for 60 hr, and were analyzed for total, insoluble, and soluble ${\beta}$-glucan contents. The total ${\beta}$-glucan content in raw barley was 3.96%, and the content was reduced during malting. The total ${\beta}$-glucan contents of the poorly- and well-modified malts were 1.02% and 0.18%, respectively. After 4 days of germination, approximately 95% of the ${\beta}$-glucan present in the barley was degraded. A significantly higher proportion of water-soluble ${\beta}$-glucan was found in the well-modified malt, suggesting that ${\beta}$-glucan solubility was dependent on cell wall modifications in the malt (${\beta}$-glucan breakdown). The proportion of water-soluble ${\beta}$-glucan was also affected by the extraction temperature. The two differently modified malts were mashed isothermally at 45, 55, 65, and 75oC for 2 hr. An increasing mashing temperature resulted in increased viscosity for the wort and the resulting beer. The viscosity of the wort from the well-modified malt was significantly low, due to its low initial malt ${\beta}$-glucan with increased solubility as well as a presumably sufficient ${\beta}$-glucanase activity during mashing.

The Synergistic Action of the AL-Protease from Arthrobacter luteus on the Lysis of Yeast Cell Walls (Arthrobacter luteus가 생산하는 AL-Protease의 효모세포벽 용해 촉진작용)

  • Oh, Hong-Rock;Funatsu, Masaru
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.4
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    • pp.401-408
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    • 1985
  • The yeast cell wall lytic action of the alkaline AL-protease, which was found out of the crude Zymolyase that a kind of yeast cell wall lytic $endo-{\beta}-1$, 3-glucanase produced from Arthrobacter luteus, was investigated with the viable cells of S. sake and it's cell wall preparation. AL-protease on the lysis of the viable yeast cells showed very low activities with the alone, but the lytic activities were highly increased with the combination of AL-protease and Zymolyase. On the stepwise treatment of the viable yeast cells with AL-protease and Zymolyase, the cells were lysed highly only by the course having a treatment with Zymolyase after pretreatment with AL-protease. Thus synergistic action of AL-protease was not observed with any some commercial enzymes, known as a type of alkaline and serine protease such as AL-protease, and was also found to be affected greatly by the culture conditions and species of the yeast tested. AL-protease caused the release of some peptide and a lot of sugar from the cell wall preparation, but could not lysed the cell wall more than 66%. Whereas Zymolyase could lysed the cell walls almost completely with alone. On the basis of these results, the synergistic action of AL-protease on the lysis of S. sake cells is hypothesized that at first AL-protease bind to the yeast cell surface layer consisting of mannan and protein, and then changes their conformation to facilitate the penetration of Zymolyase from the outside to the inside framework layer constituted of alkali insoluble ${\beta}-1,\;3-glucan$.

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Immune-Enhancing Alkali-Soluble Glucans Produced by Wild-Type and Mutant Saccharomyces cerevisiae

  • Ha Chang-Hoon;Lim Ki-Hong;Jang Se-Hwan;Yun Cheol-Won;Paik Hyun-Dong;Kim Seung-Wook;Kang Chang-Won;Chang Hyo-Ihl
    • Journal of Microbiology and Biotechnology
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    • v.16 no.4
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    • pp.576-583
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    • 2006
  • The alkali-soluble glucan of the yeast cell wall contains $\beta-(1,3)-$ and (1,6)-D-linkages and is known to systemically enhance the immune system. In the previous study [6], in order to isolate cell wall mutants, a wild-type strain was mutagenized by exposure to ultraviolet light, and the mutants were then selected via treatment with laminarinase $(endo-\beta-(1,3)-D-glucanase)$. The mass of alkali- and water-soluble glucans produced by the mutant was measured to be 33.8 mg/g of the dry mass of the yeast cell. Our results showed that the mutants generated the amount of alkali-soluble glucan 10-fold higher than that generated by the wild-type. Structural analysis showed that the alkali-soluble glucan from the mutants was associated with a higher degree of $\beta-(1,6)-D-linkage$ than was observed in conjunction with the wild-type. Yeast cell wall $\beta-glucan$ was shown to interact with macrophages via receptors, thereby inducing the release of tumor necrosis factor alpha $(TNF-\alpha)$ and nitric oxide. Alkali-soluble $\beta-glucans$, both from water-soluble and water-insoluble glucan, exhibited a higher degree of macrophage activity with regard to both the secretion of tumor necrosis factor alpha $(TNF-\alpha)$ and nitric oxide and direct phagocytosis, than did the positive control ($1{\mu}g$ of lipopolysaccharide).

Preparation and Analysis of Yeast Cell Wall Mannoproteins, Immune Enhancing Materials, from Cell Wall Mutant Saccharomyces cerevisiae

  • Ha Chang-Hoon;Yun Cheol-Won;Paik Hyun-Dong;Kim Seung-Wook;Kang Chang-Won;Hwang Han-Joon;Chang Hyo-Ihl
    • Journal of Microbiology and Biotechnology
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    • v.16 no.2
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    • pp.247-255
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    • 2006
  • Yeast cell wall matrix particles are composed entirely of mannoprotein and ${\beta}-glucan$. The mannoproteins of yeast cell wall can systemically enhance the immune system. We previously purified and analyzed alkali-soluble ${\beta}-glucans$ [${\beta}$-(1,3)- and ${\beta}$-(1,6)-glucans] [10]. In the present study, a wild-type strain was first mutagenized with ultraviolet light, and the cell wall mutants were then selected by treatment with 1.0 mg/ml laminarinase (endo-${\beta}$-(1,3)-D-glucanase). Mannoproteins of Saccharomyces cerevisiae were released by laminarinase, purified by concanavalin-A affinity and ion-exchange chromatography. The results indicated that the mutants yielded 3-fold more mannoprotein than the wild-type. The mannoprotein mass of mutant K48L3 was 2.25 mg/100 mg of yeast cell dry mass. Carbohydrate analysis revealed that they contained mannose, glucose, and N-acetylglucosamine. Saccharomyces cerevisiae cell wall components, mannoproteins, are known to interact with macrophages through receptors, thereby inducing release of tumor necrosis factor alpha ($TNF-{\alpha}$) and nitric oxide. Mannoprotein tractions in the present study had a higher macrophage activity of secretion of $TNF-{\alpha}$ and nitric oxide and direct phagocytosis than positive control ($1{\mu}g$ of lipopolysaccharide). In particular, F1 and F3 fractions in mannoproteins of K48L3 enhanced and upregulated the activity of nitric oxide secretion and macrophage phagocytosis by approximately two- and four-fold, respectively.

Study on the Anti-HT-29 Human Colon Cancer Activity of $\beta$-Glucans and Their Enzymatically Hydrolyzed Oligosaccharides from Agalicus blazei Murill (아가리쿠스로부터 분리한 $\beta$-glucan과 그 올리고당류의 HT-29 인체 대장암 세포에 대한 항암 활성에 관한 연구)

  • Lee, Mi-Young;Kim, Ki-Hoon;Kim, Yea-Woon;Chang, Hun-Gil;Lee, Dong-Seok
    • Korean Journal of Microbiology
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    • v.42 no.4
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    • pp.319-325
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    • 2006
  • [ $\beta$ ]-Glucans (AG) were prepared from Agaricus blazei cultured in the medium fortified with the roots of Pueraria spp. by repeated extraction with hot water, gel filtration chromatography and DEAE ion exchange chromatography. Oligosaccharides (AO) were derived from the hydrolysis of AG by an endo-$\beta$-(1$\rightarrow$6)-glucanase from Bacillus megaterium. The anti-HT-29 human colon cancer activity of AG or AO was investigated using MTT assay, apoptosis assay, cell cycle analysis, and cDNA microairay. AG and AO both inhibited proliferation and growth of HT-29 cells, and stimulated apoptosis of the cells in a dose-dependent manner. In cell cycle analysis, treating HT-29 cells with AG or AO resulted in the increase of cells in the G0 (sub-G1) and G1 phase. Especially, AO was more effective in inducing G0/G1 cell cycle arrest than AG. To screen the genes involved in the increase of apoptosis, the gene expression profile of the HT-29 cells treated with AO was examined by cDNA microarray. While several genes involved in cell cycle progression (CCND2 and CDK2) were down-regulated, many genes involved in apoptosis (TNFSF9, TNFRSF9, FADD, CASP8, BAD, CRADD, CASP9 etc), cell cycle inhibitor (CDKN2A), immune response (IL6, IL18, IL6R etc), and tumor suppressor (CEACAM1, TP53BP2, IRF1, and PHB) were up-regulated. These results suggest that AO could inhibit the proliferation and growth of HT-29 cells by G0/G1 cell cycle arrest and induction of apoptosis.

Disease Development in Resistant Tobacco Plants Infected with Tobacco Mosaic Virus and Expression of Pathogenesis-Related Genes in Different Temperature Conditions (TMV에 감염된 저항성 담배 식물체의 온도 조건에 따른 병발생 특성 및 PR 유전자 발현)

  • 김영호;박은경;윤해근;최도일;채순용;강신웅
    • Journal of the Korean Society of Tobacco Science
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    • v.18 no.1
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    • pp.21-29
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    • 1996
  • Tobacco plants resistant (cvs. Xanthi-nc and Samsun-NN) and susceptible (cv. NC 82) to tobacco mosaic virus (TMV) were inoculated with TW to obtain basic information about the characteristics of resistance expression in tobacco plants by examining the viral populations, symptom development and gene expression of pathogenesis-related proteins (PR-proteins) such as PR-1 and $\beta$-1, 3-glucanase in different temperature conditions. TMV populations in resistant plants increased more at 37$^{\circ}C$ than at 27$^{\circ}C$, while the viral populations increased continuously and were not significantly influenced by the temperature conditions in the susceptible tobacco plants. Infection sites of resistant tobacco leaves were remarkably expanded in proportion with increased time at the high temperature.

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Multicarbohydrase Enzymes for Non-ruminants

  • Masey O'Neill, H.V.;Smith, J.A.;Bedford, M.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.2
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    • pp.290-301
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    • 2014
  • The first purpose of this review is to outline some of the background information necessary to understand the mechanisms of action of fibre-degrading enzymes in non-ruminants. Secondly, the well-known and understood mechanisms are described, i) eliminating the nutrient encapsulating effect of the cell wall and ii) ameliorating viscosity problems associated with certain Non Starch Polysaccharides, particularly arabinoxylans and ${\beta}$-glucans. A third, indirect mechanism is then discussed: the activity of such enzymes in producing prebiotic oligosaccharides and promoting beneficial cecal fermentation. The literature contains a wealth of information on various non starch polysaccharide degrading enzyme (NSPase) preparations and this review aims to conclude by discussing this body of work, with reference to the above mechanisms. It is suggested that the way in which multi- versus single-component products are compared is often flawed and that some continuity should be employed in methods and terminology.