• Title/Summary/Keyword: $\beta$-Glycosidase

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Characterization of Two Algal Lytic Bacteria Associated with Management of the Cyanobacterium Anabaena flos-aquae

  • Kim, Jeong-Dong;Lee, Choul-Gyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.5
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    • pp.382-390
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    • 2006
  • Various microorganisms were isolated from the surface waters and sediments of eutrophic lakes and reservoirs in Korea to enable an investigation of bacteria having algal lytic activities against Anabaena flos-aquae when water blooming occurs and to study enzyme profiles of algal lytic bacteria. Two bacterial strains, AFK-07 and AFK-13, were cultured, characterized and identified as Acinetobacter johnsonii and Sinorhizobium sp., respectively. The A. johnsonii AFK-07 exhibited a high level of degradatory activities against A. flos-aquae, and produced alginase, caseinase, lipase, fucodian hydrolase, and laminarinase. Moreover, many kinds of glycosidase, such as ${\beta}-galactosidase,\;{\beta}-glucosidase,\;{\beta}-glucosaminidase,\;and\; {\beta}-xylosidase$, which hydrolyzed ${\beta}-O-glycosidic$ bonds, were found in cell-free extracts of A. johnsonii AFK-07. Other glycosidases such as ${\alpha}-galactosidase,\;{\alpha}-N-Ac-galactosidase,\;{\alpha}-mannosidase,\; and\;{\alpha}-L-fucosidase$, which cleave ${\alpha}-O-glycosidic$ bonds, were not identified in AFK-07. In the Sinorhizobium sp. AFK-13, the enzymes alginase, amylase, proteinase (caseinase and gelatinase), carboxymethyl-cellulase (CMCase), laminarinase, and lipase were notable. No glycosidase was produced in the AFK-13 strain. Therefore, the enzyme system of A. johnsonii AFK-07 had a more complex mechanism in place to degrade the cyanobacteria cell walls than did the enzyme system of Sinorhizobium sp. AFK-13. The polysaccharides or the peptidoglycans of A. flos-aquae may be hydrolyzed and metabolized to a range of easily utilized monosaccharides or other low molecular weight organic substances by strain AFK-07 of. A. johnsonii, while the products of polysaccharide degradation or peptidoglycans were more likely to be utilized by Sinorhizobium sp. AFK-13. These bacterial interactions may offer an alternative effective approach to controlling the water choking effects of summer blooms affecting our lakes and reservoirs.

Biotransformation of Ginsenoside by Lactobacillus brevis THK-D57 Isolated from Kimchi (김치에서 분리한 Lactobacillus brevis THK-D57에 의한 인삼 사포닌의 생물학적 전환)

  • Yi, Eun-Ji;Lee, Jung-Min;Yi, Tae-Hoo;Cho, Seok-Cheol;Park, Yong-Jin;Kook, Moo-Chang
    • The Korean Journal of Food And Nutrition
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    • v.25 no.3
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    • pp.629-636
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    • 2012
  • Ginsenosides, ginseng saponin, are the principal components responsible for the pharmacological and biological activities of ginseng. In order to improve absorption and biological activities, the biotransformation of major ginsenoside to minor ginsenoside, as the more active compound, is required. In this study, we isolated Lactobacillus brevis THK-D57, which has high ${\beta}$-glycosidase activity, from Kimchi. The major ginsenoside Rb1 was converted to the minor ginsenoside 'compound K' during the fermentation of L. brevis THK-D57. The results propose that the biotransformation pathway to produce compound K is as follows: ginsenoside $Rb_1{\rightarrow}ginsenoside$ $Rd{\rightarrow}ginsenoside$ $F_2{\rightarrow}ginsenoside$ compound K.

Effects of IAA on the Elongation and Cell Wall Glycosidase Activities in Excised Rape (Brassica napus L. cv. Yongdang) Hypocotyl Segments (유채 하배축 분절의 신장과 세포벽 분해효소의 활성에 미치는 IAA의 효과)

  • Jun, Sung-Soo
    • Journal of Plant Biology
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    • v.27 no.2
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    • pp.43-50
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    • 1984
  • Effects of IAA on the elongation and cell wall hlysocidase activities were investigated in excised rape (Brassica napus L. cv. Yongdang) hypocotyl segments. IAA promoted the elongation of rape hypocotyl segments. In rape hypocotyls, the first 10-mm segments from the hook exhibited maximal elongation and the capacity of elongation was gradually decreased with increasing distance of each 10-mm from the hook. A good correlation has been obtained between the magnitude of endogenous growth and the activities of $\alpha$, $\beta$-glucosidase and $\alpha$, $\beta$-galactosidase. However, exogenous application of IAA did not seem to enhance the tissue with IAA resulted in acidification of the incubation medium. From these data, we can conclude that IAA seems to enhance elongation of the tissue segments, at least in part, by releasing hydrogen ion into cell wall, some of which may participate in the cell wall extension process, but does not seem to trigger the activation of $\alpha$, $\beta$-glucosidase and $\alpha$, $\beta$-galactosidase.

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Purification and Characterization of an α-D-Galactosidase from Grape Berry

  • Kang, Han-Chul;Kim, Tae-Su
    • Journal of Applied Biological Chemistry
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    • v.43 no.3
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    • pp.141-146
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    • 2000
  • Glycosidase activities were tested from the grape berries, Vitis labruscana B. Takasumi. Among various glycosidases, $\alpha$-D-galactosidase was found to be the most active in the flesh and other glycosidases were considerably active in the order of the following: $\alpha$-D-mannosidase>$\alpha$-D-glucosidase>$\beta$-D-glucosidase>$\beta$-D-galactosidase. In the seeds, $\alpha$-D-glucosidase activity was the highest and other glycosidases such as $\alpha$-D-galactosidase, $\beta$-D-glucosidase, and $\beta$-D-galactosidase were still significantly active. The $\alpha$-D-galactosidase in the grape flesh was purified over 83-folds through salting-out with $(NH_4)_2SO_4$ and a series of chromatographies employing Sephadex G-50, Octyl-Sepharose, Q-Sepha- rose, and Biogel P-100. The enzyme was a monomer of 45 kDs as determined through SDS-PAGE and Sephacryl S-200 chromatography. The purified enzyme showed a preference of $\alpha$-D-galactose to $\beta$-D-galactose as a substrate about 5.4 times. Sulfhydryl specific reagents such as N-ethylmaleimide and iodoacetamide significantly inhibited the enzyme activity to the extents of 48 and 52% of its initial activity, respectively. The optimumpH range of $\alpha$-D-galactosidase was around 6.5-7.0. The enzyme activity increased by 46% in the presence of 1mM $Fe^{2+}$.

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Effects of ${\beta}-Glucan$ from Agaricus blazei Murill on Blood Glucose and Lipid Composition in db/db Mice (db/db 마우스에서 아가리쿠스 버섯 ${\beta}-Glucan$이 혈당과 지질성분에 미치는 영향)

  • Choi, Jung-Mi;Koo, Sung-Ja
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1418-1425
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    • 2000
  • Obesity and diabetes mellitus are associated with common pathogenic mechanism, and ${\beta}-glucan$ of Agaricus blazei Murill is potent inhibitor of intestinal ${\alpha}-glycosidase$ and inhibit the digestion of starch and sucrose in the small intestine. In this studies, there was observed the anti-hyperglycemic effect in obese diabetic mice(C57BLKsJ db/db), which were supplied Agaricus and Acarbose for 5 weeks. In db/db mice, food intake and body weight gain were decreased significantly in Agaricus groups(p<0.05). Also these group exhibited lower fasting serum glucose level compared with control group. HbA1c level, triglyceride level, total cholesterol level, HDL cholesterol level, LDL cholesterol level and VLDL cholesterol level were lowered in db/db mice. The activity of disaccharidases on proximal and distal segments of small intestine was decreased. In conclusion, it was assumed that ${\beta}-glucan$ of Agaricus blazei Murill has anti-hyperglycemic and anti-obesitic effects by reducing food intake and body weight gain, and also decreasing serum glucose and lipid level through inhibiting the activity of small intestinal disaccharidases.

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Studies on Adivitie of $\beta$-Glucuronidase and Several Glycosidases of the Castrated Rat Epi-didymis Treated with Testosterone and Dibutyryl cAMP and the Cell Types of Epididymal Epithelium (Testosterone과 dibutyryl cyclic AMP가 거세한 흰쥐 부정소의 $\beta$ -glucosidase와 몇가지 glycosidase 활성에 미치는 영향 및 부정소 상피세포의 여러 유형에 관한 연구)

  • 최임순;정경순
    • The Korean Journal of Zoology
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    • v.32 no.3
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    • pp.290-303
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    • 1989
  • The activities of $\beta$-glucosidase, $\beta$-glucuronidase and N-acetyl-$\beta$-glucosaminidase were measured to investigate the relationships of them to sexual maturity. Peritoneal injections of testosterone and dibutyryl cAMP to rats were carried out. As a result, the activities of $\beta$-glucosidase and N-acetyl-$\beta$-glucosaminidase were significantly decreased from the third day and that of P -glucurondiase on the seventh day in the castrated groups. In addition, ihe activities of these three enzymes were significantly increased in the testosterone treated groups for 7 days. In case of dbcAMP injection, the activities of these three enzymes were similar to those of castrated groups or had a tendency to be decreased. On electron microscopic examination, principal cells, basal cells and narrow cells were observed in all regions of epididymis. Principal cells were general forms of columnar epithelial cells. Narrow cells had a number of small vesicles and light cells showed low electron density in comparison to other epithelial cells in cauda epididymis. Halo cells were migrating leucocytes btween epithelial cells.

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One-Step Enzymatic Synthesis of Blue Pigments from Geniposide for Fabric Dyeing

  • Cho, Y.J.;Kim, S.Y.;Kim, J.;Choe, E.K.;Kim, S.I.;Shin, H.J.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.3
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    • pp.230-234
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    • 2006
  • In this study, we describe a one-step chemoenzymatic reaction for the production of natural blue pigments, in which the geniposide from Gardenia extracts is transformed by glycosidases to genipin. Genipin is then allowed to react with amino acids, thereby generating a natural blue pigment. The ${\beta}-glycosidases$, most notably Isolase (a variant of ${\beta}-glucanase$), recombinant ${\beta}-glycosidases$, Cellulase T, and amylases, were shown to hydrolyze geniposide to produce the desired pigments, whereas the ${\alpha}-glycosidases$ did not. Among the 20 tested amino acids, glycine and tyrosine were associated with the highest dye production yields. The optimal molar ratio of geniposide to glycine, two reactants relevant to pigment production, was unity The natural blue pigments produced in this study were used to dye cotton, silk, and wool. The color yields of the pigments were determined to be significantly higher than those of other natural dyes. Furthermore, the color fastness properties of these dyes were fairly good, even in the absence of mordant.

Isolation, Identification, and Characterization of Pichia guilliermondii K123-1 and Candida fermentati SI, Producing Isoflavone β-Glycosidase to Hydrolyze Isoflavone Glycoside Efficiently, from the Korean Traditional Soybean Paste

  • Kim, Won-Chan;So, Jai-Hyun;Kim, Sang-In;Shin, Jae-Ho;Song, Kyung-Sik;Yu, Choon-Bal;Kho, Yung-Hee;Rhee, In-Koo
    • Journal of Applied Biological Chemistry
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    • v.52 no.4
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    • pp.163-169
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    • 2009
  • A total of 155 microbial strains were isolated from the Korean traditional soybean paste based on their morphological features on the growth of agar plate. Among the isolated strains, a total of 28 strains were capable of hydrolyzing isoflavone glycoside to isoflavone aglycone efficiently in the soybean paste. Finally, two strains, K123-1 and SI, were selected because of their resistance to 15% NaCl and ability to convert isoflavone glycoside to isoflavone aglycone efficiently during the fermentation of soybean paste. The isolated strains K123-1 and SI were identified to be Pichia guilliermondii and Candida fermentati, respectively, using the partial 26S rDNA sequence analysis and phylogenic analysis. Pichia guilliermondii K123-1 and Candida fermentati SI converted daidzin to daidzein up to 96% and 95%, respectively, and genistin to genistein up to 92% when soybean pastes were fermented at $30^{\circ}C$ for 20 days with a single isolated strain. Pichia guilliermondii K123-1 and Candida fermentati SI were able to grow in the presence of 15% NaCl on both liquid medium and agar plate. We think that Pichia guilliermondii K123-1 and Candida fermentati SI might be one of good candidates for making functional soybean paste because they are isolated from the Korean traditional soybean paste and have a good ability to convert isoflavone glycosides to isoflavone aglycones and a high salt tolerance.

Single-Chain Fv Fragment of Catalytic Antibody 4f4f with Glycosidase Activity: Design, Expression, and Purification

  • Jang, Chang-Hwan;Chung, Hyun-Ho;Yu, Jae-Hoon;Chang, Yung-Jin;Kim, Hyong-Bai;Paek, Se-Hwan;Shin, Dong-Hoon;Kim, Kyung-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.376-380
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    • 1999
  • Constructs, encoding a single-chain variable fragment of a catalytic antibody 4f4f (scFv-4f4f) with glycosidase activity, were made by combining the coding sequences for the heavy and light chain variable domains with a sequence encoding a linker (GGGGS). Using three different plasmid systems, single-chain antibodies were expressed separately in Escherichia coli, demonstrating significant differences in the expression level and amounts in soluble form of the recombinant protein. The protein expression from pET3a-scFv-4f4f was up to 20% of the total soluble proteins and, more importantly, the proteins were mostly found in a soluble form. An SDS-PAGE analysis of the purified single-chain proteins, yielding higher than 5mg from a 1-1 culture, showed a single band corresponding to its molecular weight of 29,100. A preliminary study shows that the expressed scFv-4f4f is catalytically active. The catalytic parameters for the hydrolysis of p-nitrophenyl-$\beta$-D-glucopyranoside by scFv-4f4f are being investigated.

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