• Title/Summary/Keyword: Glucan

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A MALDI-MS-based Glucan Hydrolase Assay Method for Whole-cell Biocatalysis

  • Ahn, Da-Hee;Park, Han-Gyu;Song, Won-Suk;Kim, Seong-Min;Jo, Sung-Hyun;Yang, Yung-Hun;Kim, Yun-Gon
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.69-77
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    • 2019
  • Screening microorganisms that can produce glucan hydrolases for industrial, environmental, and biomedical applications is important. Herein, we describe a novel approach to perform glucan hydrolase screening-based on analysis of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) spectra-which involves degradation of the oligo- and polysaccharides. As a proof-of-concept study, glucan hydrolases that could break down glucans made of several glucose units were used to demonstrate the MALDI-MS-based enzyme assay. First, the enzyme activities of ${\alpha}$-amylase and cellulase on a mixture of glucan oligosaccharides were successfully discriminated, where changes of the MALDI-MS profiles directly reflected the glucan hydrolase activities. Next, we validated that this MALDI-MS-based enzyme assay could be applied to glucan polysaccharides (i.e., pullulan, lichenan, and schizophyllan). Finally, the bacterial glucan hydrolase activities were screened on 96-well plate-based platforms, using cell lysates or samples of secreted enzyme. Our results demonstrated that the MALDI-MS-based enzyme assay system would be useful for investigating bacterial glucoside hydrolases in a high-throughput manner.

Effect of Various β-1,3-glucan Supplements on the Performance, Blood Parameter, Small Intestinal Microflora and Immune Response in Laying Hens (β-Glucan 제제들이 산란계의 생산성, 혈액 성상과 소장내 미생물 균총 및 면역 체계에 미치는 영향)

  • Park, K.W.;Rhee, A.R.;Lee, I.Y.;Kim, M.K.;Paik, I.K.
    • Korean Journal of Poultry Science
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    • v.35 no.2
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    • pp.183-190
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    • 2008
  • This study was conducted to investigate the effect of feeding diets supplemented with ${\beta}-glucan$ products on the performance, small intestinal microflora and immune response in laying hens. The ${\beta}-glucan$ products used in the experiment were $BetaPolo^{(R)}$ ; soluble ${\beta}-glucan$ of microbial cell wall origin, $HiGlu^{(R)}$ ; microbial cell wall origin, $OGlu^{(R)}$ ; oat origin, $BGlu^{(R)}$ ; barley origin. A total of 720 Hy-Line Brown laying hens of 40wks old were divided into 5 dietary treatments : T1 ; Control( C), T2 ; $BetaPolo^{(R)}$, T3 ; $HiGlu^{(R)}$, T4 ; $OGlu^{(R)}$, T5 ; $BGlu^{(R)}$. Each treatment was replicated 4 times with 36 birds/replicate housed in 2 bird cages, and arranged according to completely randomized block design. Feeding trial lasted 40ds under 16 h lighting regimens. There were significant differences among treatments in hen-house egg production feed intake and feed conversion. HiGlu treatment was significantly higher than OGlu treatments in hen-house egg production. ${\beta}-glucan$ supplemented treatments were lower than the control in feed intake and feed conversion ratio. All ${\beta}-glucan$ supplemented treatments were significantly higher than the control in eggshell strength. Eggshell color and Haugh unit tended to be lower in the supplemented group than the control. IgY concentration was not significantly affected by treatments. At $5^{th}$ week of experiment, however, IgY concentration tended to increase in the supplemented groups. Among the leucocytes parameters, WBC, heterophil, lymphocytes, monocyte and eosinophil concentration were lower in the supplemented groups than those of the control. Among erythrocytes, HCT(hematocrit) and MCV(mean corpuscular volume) were significantly affected by treatment. MCV of supplemented groups were higher than that of the control. Immunoglobulin concentrations in the birds were not significantly different among treatments. However, IgA concentration tended to be low in the supplemented groups than the control. The cfu of small intestinal microflora were not significantly different among treatments, but that of Cl. perfringens tended to be lower than the control. The result of this experiment indicateted that feeding ${\beta}-glucan$ to laying hens improve feed conversion ratio and eggshell strength. Also intestinal microflora and immune responses are modified.

Preventive Mechanisma of ${\beta}$-Glucan on the Experimental Atherosclerosis in Rats (실험적으로 동맥경화증을 유발시킨 랫드에서 ${\beta}$-Glucan의 동맥경화 방어기전에 대한 연구)

  • 김형욱;이영순
    • Biomolecules & Therapeutics
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    • v.1 no.1
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    • pp.109-120
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    • 1993
  • Mechanisms for the hypocholesterolemic effetcs of $\beta$-glucan remain unclear. Rats were divided into 3 groups; normal control group, atherogenic group(oral administration of cholesterol 40 mg/kg/day and vitamin $D_2$320,000 IU/kg/day),${\beta}$-glucan treatment group(oral administration of atherogenic treatment and ${\beta}$-glucan 0.135 g/kg/day). The ${\beta}$-glucan treatment group showed moderate increases of serum lipids concentration compared with atherogenic group. In histopathological examination, aortas showed no critical lesions. The total fecal neutral sterols and bile acids excreted for 6 days was increased compared with both normal and atherogenic group. These result\ulcorner suggest that mechanisms for the hypocholesterolemic effect of ${\beta}$-glucan on rats were due to the inhibition of cholesterol absorption in the intestinal lumen and acceleration of cholesterol catabolism in the liver.

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Development of Soluble Glucan Production Process with Continuous Stirred Tank Fermentor (연속 발효조를 이용한 soluble glucan 생산 공정 개발)

  • Moon, Chan-Jun;Lee, Jung-Heon
    • Journal of Life Science
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    • v.16 no.6
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    • pp.938-941
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    • 2006
  • Continuous fermentation process for the production of soluble glucan using mutant Agrobacterium sp. ATCC31750 has been developed in this research. When the concentration of soluble glucan was higher than 6 g/l, the viscosity of the fermented broth was too high and it needs complex separation process to separate from culture broth. Mathematical models which describe the cell growth and glucan production was developed and they kinetic parameters were estimated with experimental data. They are used for the optimization of continuous fermentation process and calculate optimal dilution rate for easy separation of glucan 4 g/l. With continuous fermentation, glucan production rate was increased 1.8 times more than that with batch fermentation.

Physicochemical Properties of Depolymerized Barley β-Glucan by Alkali Hydrolysis (알칼리 가수분해에 따른 보리 β-Glucan의 이화학적 특성)

  • Lee, Sang-Hoon;Jang, Gwi-Yeong;Kim, Kee-Jong;Lee, Mi-Ja;Kim, Tae-Jip;Lee, Jun-Soo;Jeong, Heon-Sang
    • The Korean Journal of Food And Nutrition
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    • v.26 no.3
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    • pp.601-607
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    • 2013
  • This study was performed to investigate the changes of total and soluble ${\beta}$-glucan contents, purity and physicochemical characteristics of alkali hydrolyzed barley varieties: Saessalbori (SSB), Saechalssalbori (SCSB) and Hinchalssalbori (HCSB). The barleys were hydrolyzed at different concentrations of sodium hydroxide (0.2~1.0 N) for 12 hours. Total ${\beta}$-glucan contents of raw SSB, SCSB and HCSB were 8.40, 7.77 and 8.28%, and soluble ${\beta}$-glucan contents were 4.80, 4.16 and 4.61%, respectively. The total ${\beta}$-glucan contents after alkali hydrolyzed at 1.0 N NaOH were 7.54, 6.89 and 7.54%, also soluble ${\beta}$-glucan contents were 4.82, 4.30 and 4.55%, respectively. The degree of purity of soluble ${\beta}$-glucan in SSB, SCSB, and HCSB were 35.79, 30.91 and 33.90%, respectively. They were increased to 74.02, 75.28 and 81.41% after hydrolyzed at 1.0 N NaOH, respectively. The molecular weight and viscosity of soluble ${\beta}$-glucan solutions were decreased as sodium hydroxide concentration was increased. The re-solubility of raw barley ${\beta}$-glucan was about 50%; however, it was increased to approximately 87% as sodium hydroxide concentration was increased.

Comparative Study of Immune-Enhancing Activity of Crude and Mannoprotein-Free Yeast-Gluean Preparations

  • Kim, Hye-Nam;Lee, Jung-Nam;Kim, Gi-Eun;Ha-Lee, Young-Mie;Kim, Chan-Wha;Sohn, Jeong-Won
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.826-831
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    • 1999
  • ${\beta}-Glucan$, one of the major cell wall components of Saccharomyces cerevisiae, is known to enhance the immune function, especially by activating macrophages. Accordingly, in an effort to develop a safe and efficient immune stimulatory agent, we prepared crude ${\beta}-glucan$ (glucan-p1) and partially purified ${\beta}-glucan$ that was free of mannoproteins (glucan-p2), and evaluated their effect on both the macrophage function and resistance to E. coli-induced peritonitis. To investigate the function of the macrophages, phagocytosis, $TNF-{\alpha}$ secretion, oxygen burst, and the expression of cytokine genes such as $IFN-{\gamma}$ and IL-12 were analyzed. Glucan-p2 markedly stimulated the macrophages with all these parameters. Glucan-p1, however, did not stimulate phagocytosis, yet it induced $TNF-{\alpha}$ secretion, oxygen burst, and the expression of $IFN-{\gamma}$ and IL-12, although less efficiently than glucan-p2. Finally, to test the in vivo protective effect of {\beta}-glucan against infection, the survival of mice from E. coli-induced peritonitis was investigated. After 24 h of the peritoneal challenge of E. coli, all of the mice treated with glucan-p2 survived whereas none survived in the control group. Glucan-p1 showed only a marginal effect in protecting the mice. These results suggest that mannoprotein-free gluean-p2, but not gluean-p1, can serve as an effective immune-stimulating agent.

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Effect of Barley ${\beta}-Glucan$ on Dynamic Viscoelasticity of Barley Starch (${\beta}-Glucan$이 보리 전분의 동적점탄성에 미치는 영향)

  • Choi, Hee-Don;Seog, Ho-Moon;Kim, Yun-Sook;Lee, Cherl-Ho
    • Korean Journal of Food Science and Technology
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    • v.35 no.6
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    • pp.1022-1027
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    • 2003
  • The effect of ${\beta}$-glucan, prepared from waxy barley, on the dynamic viscoelasticity of nonwaxy and waxy barley starch during gelatinization and gelation was studied. Although no significant effect was observed on waxy starch, there were drastic changes in the dynamic viscoelasticity of nonwaxy starch. The gelatinization onset temperature of nonwaxy starch shifted to a higher temperature and showed a drastic increase in storage modulus and loss modulus at the range of $80{\sim}90^{\circ}C$. During the gelation of nonwaxy starch, ${\beta}$-glucan increased the rate of gel formation and weakened the network of starch and amylose by prohibiting their association. Therefore, we proved that there was no specific interaction between amylose and ${\beta}$-glucan. The addition of ${\beta}$-glucan to waxy starch seemed to have no effect of waxy starch.

Comparison of β-glucan Contents of Lentinula edodes Cultivated on Sawdust according to Medium Composition and Fruiting Temperature (표고 톱밥재배에서 배지조성과 버섯발생 온도에 따른 β-glucan 함량 비교)

  • Park, Young-Ae;Bak, Won-Chull;Ka, Kang-Hyeon;Koo, Chang-Duck
    • The Korean Journal of Mycology
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    • v.44 no.4
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    • pp.296-299
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    • 2016
  • Lentinula edodes is an edible mushroom that contains a ${\beta}$-glucan called lentinan, which has antitumor and immune-enhancing properties. In the present study, the ${\beta}$-glucan contents of L. edodes mushrooms cultivated on sawdust with different nutritional supplements and fruiting temperatures were measured using a commercial ${\beta}$-glucan assay kit purchased from Megazyme (Bray, Ireland). The weight loss of sawdust media and the yield of fruiting bodies showed similar trends, but the yield was more closely associated with the nutritional supplements used than the weight loss of sawdust media was. The ${\beta}$-glucan contents of L. edodes were 39.5-42.1%, except in the bean curd refuse + $CaCl_2$ supplementation group (50.4%). Furthermore, the ${\beta}$-glucan content decreased with increasing temperatures and was 42.4% at a low fruiting temperature.

Effect of ${\beta}-Glucan$ on Gelatinization of Barley Starch (${\beta}-Glucan$이 보리 전분의 호화에 미치는 영향)

  • Choi, Hee-Don;Seog, Ho-Moon;Kim, Sung-Ran;Park, Yong-Kon;Lee, Cherl-Ho
    • Korean Journal of Food Science and Technology
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    • v.35 no.4
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    • pp.545-550
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    • 2003
  • The effect of ${\beta}-glucan$ on gelatinization of barley starch was studied. By the rapid visco-analyzer measurement, gelatinization of starch became rapid and viscosity increased largely on the RVA pattern by addition of ${\beta}-glucan$ to starch. The results of differential scanning calorimeter showed that molecular structure of starch was getting stabilized through shifting up of gelatinization temperature and increase in enthalpy by addition of ${\beta}$-glucan. X-ray diffraction pattern also showed the same results as differential scanning calorimeter. But it was revealed that addition of ${\beta}$-glucan to starch didn't affect characteristics such as microscopic observation, solubility, swelling power, and iodine binding properties during gelatinization of starch.

Studies on Rheological Characterization of Barley ${\beta}-Glucan$ [mixed-linked $(1-3),(1-4)-{\beta}-D-Glucan$] (보리 ${\beta}-Glucan$ [mixed-linked $(1-3),(1-4)-{\beta}-D-Glucan$의 리올로지 특성)

  • Kim, Mi-Ok;Cha, Hee-Sook;Koo, Sung-Ja
    • Korean Journal of Food Science and Technology
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    • v.25 no.1
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    • pp.15-21
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    • 1993
  • Crude ${\beta}-glucan$ extracted from Barley was purified by stepwise enzyme treatment (Thermostable ${\alpha}-amylase$, amyloglucosidase, protease). The Intrinsic Viscosity $[{\eta}]$ of the purified ${\beta}-glucan$ was determined by Cannon Fenske Capillary Viscometer (size 50, Cannon Instruments, State, College pa.) at different pH (2, 4, 7, 9, 11) and various salt concentration (0.01 M, 0.03 M, 0.05 M, 0.07 M, 0.1 M and 0.2 M). The $[{\eta}]$ of purified ${\beta}-glucan$ was ranged from $0.997{\sim}2.290\;dl/g$. The $[{\eta}]$ of purified ${\beta}-glucan$ at both alkali, acid condition were lower than those at pH 7. However, the alkali condition of puified ${\beta}-glucan$ solution showed less $[{\eta}]$ than the acid condition of this solution. From 0 M to 0.2 M salt concentration, the $[{\eta}]$ of purified ${\beta}-glucan$ solution was decreased to 0.03 M then increased to 0.05 M NaCl and remained constant to 0.2 M NaCl. The chain stiffness parameter of purified ${\beta}-glucan$ was not affected by temperature from $15^{\circ}C$ to $65^{\circ}C$. The shear rates of various ${\beta}-glucan$ conditions were determined by Bohlin Rheometer (Lund, Sweden). The ${\beta}-glucan$ concentration of 1.0 g/dl and 2.0 g/dl behaved as Newtonian fluid. However, above the concentration of 3.0 g/dl ${\beta}-glucan$ solution, it showed thixotropic and psedoplastic characteristics. Barley ${\beta}-glucan$ appears a damping at 0.5 frequency for the 4.0 g/dl solution. Below 0.5 frequency, it appears a viscous behavior property and above 0.5 frequency, it appears a elastic behavior property.

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