• Title/Summary/Keyword: Glycosidase

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Total synthesis of 1,4-Dideoxy-1,4-Imino-D-Arabinitol(DABl)

  • Kim, In-Su;Jung, Young-Hoon
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.178.2-178.2
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    • 2003
  • Naturally occurring sugar mimics with a nitrogen in the ring are classified into five structural classes: polyhydroxylated pyrrolidines, piperidines, indolizidines. pyrrolizidine, and nortropanes. Glycosidase are involved in a wide range of important biological processes, such as intestinal digestion, post-translational processing of glycoproteins and the lysosomal catabolism of glycoconjugate. The realization that alkaloidal sugar mimics might have enormous therapeutic potential in many diseases such as viral infection, cancer and diabetes has led to increasing interest and demand for these compounds. (omitted)

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Enzymatic Characterization and Substrate Specificity of Thermostable $\beta-Glycosidase$ from Hyperthermophilic Archaea, Sulfolobus shibatae, Expressed in E. coli

  • Park, Na-Young;Cha, Jae-Ho;Kim, Dae-Ok;Park, Cheon-Seok
    • Journal of Microbiology and Biotechnology
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    • v.17 no.3
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    • pp.454-460
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    • 2007
  • Enzymatic properties and substrate specificity of recombinant $\beta-glycosidases$ from a hyperthermophilic archaeon, Sulfolobus shibatae (rSSG), were analyzed. rSSG showed its optimum temperature and pH at $95^{\circ}C$ and pH 5.0, respectively. Thermal inactivation of rSSG showed that its half-life of enzymatic activity at $75^{\circ}C$ was 15 h whereas it drastically decreased to 3.9 min at $95^{\circ}C$. The addition of 10 mM of $MnCl_2$ enhanced the hydrolysis activity of rSSG up to 23% whereas most metal ions did not show any considerable effect. Dithiothreitol (DTT) and 2-mercaptoethanol exhibited significant influence on the increase of the hydrolysis activity of rSSG rSSG apparently preferred laminaribiose $(\beta1\rightarrow3Glc)$, followed by sophorose $(\beta1\rightarrow2Glc)$, gentiobiose $(\beta1\rightarrow6Glc)$, and cellobiose $(\beta1\rightarrow4Glc)$. Various. intermolecular transfer products were formed by rSSG in the lactose reaction, indicating that rSSG prefers lactose as a good acceptor as well as a donor. The strong intermolecular transglycosylation activity of rSSG can be applied in making functional oligosaccharides.

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.

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.

Characteristics of the Cell Wall Lytic Enzyme of Anabaena cylindrica from Penicillium oxalicum(HCLF-34) (Penicillium oxalicum(HCLF-34)으로부터 분비되는 Anabaena cylindrica 세포벽 분해효소의 특성)

  • 현성희;최영길
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.231-236
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    • 1999
  • The fuugus(Penicil1ium oralicum; HCLF-34) secreted the cyanobacteria lytic enzyme which had a molecular weight of about 22 kDa, a optimum temperature of $20^{\circ}C$, a optimum pH of 3.5, and a temperature-stable up to $50^{\circ}C$. The chemical ions such as sodium, potassium, barium, magnesium. and mangan ions appeared positive activity. but calcium, iron, copper ions, EDTA, and PMSF displayed negative activity: this results were the same as the characterilics of other cell wall lytic enzymes. This extracellular enzyme showed lytic aclivily against SDS-insoluble peptidoglycan of Anabaenrr cylinrlrica. The cell wall lylic enzyme of Penicilliurn oxalicum(HCLF-34) seemed to be glycosidase-like enzyme in the fact that ihe concentration of rcducing sugar was increased when the peptidoglycan of Anabaena qlinrlricn md Micrococcus luteus reacted with this enzyme

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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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Development of An Anti-Diabetic Functional Drink (I) - Screening of the Manufacturing of Mulberry Leaf Extract - (상엽 추출물을 이용한 항당뇨 음료의 개발 (I) - 상엽 추출물의 제제화 탐색 -)

  • 구성자;윤기주;김근풍
    • Journal of the East Asian Society of Dietary Life
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    • v.12 no.5
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    • pp.364-369
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    • 2002
  • Mulberry leaves are believed to be a potent inhibitor of intestinal $\alpha$-glycosidase and the digestion of sucrose in the small intestine. Mulberry leaves are also blown to help prevent the postprandial hyperglycemia. The objective of this study was to elucidate the functionality and anti-hyperglycemic effect of mulberry leaves, and to develop a functional drink using mulberry leaf, silk peptide and oriental medicine. Several mixtures of mulberry lear extract silk peptide and oriental medicine were made for the purpose of manufacture and formulation of products. These mixtures were tested to check the $\alpha$-glycosidase inhibition effect to find the best formula. The optimum conditions for a mulberry leaf hot water extract were that the size of leaf was under 3mm, the amount of leaf needed 50~100 g/L, extraction temperature 9$0^{\circ}C$ and extraction time 2 hr. The yield of extraction was 20~25%. The anti-hyperglycemic effect of mulberry leaves was at maximum when the concentrations of mulberry leaf extract and silk peptide were 1% and 0.1%, respectively.

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Blood Glucose Lowering Activity and Mechanism of Supungsungihyan (SPSGH) in db/db Mouse (db/db 마우스에서 수풍순기환의 혈당강하 활성 및 기전연구)

  • 이성현;안세영;두호경;정성현
    • Biomolecules & Therapeutics
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    • v.7 no.4
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    • pp.335-341
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    • 1999
  • Antidiabetic activity and mechanism of Supungsungihyan(SPSGH) were examined in db/db mice, which is a spontaneously hyperglycemic, hyperinsulinemic and obese animal model. SPSGH and acarbose were administered orally for 4 weeks. Fasting and non-fasting serum glucose, glycated hemoglobin and trig-lyceride of SPSGH treated group were all reduced when compared with those of db/db control group. At 12th week after birth, SPSGH increased an insulin secretion although statistic significance was not seen. Total activities of sucrose, maltase and lactase in SPSGH treated group were not significantly different from those in db/db control. On the other hand, sucrase and maltase activities in acarbose treated groups were increased. Effect of SPSGH on mRNA expression of glucose transporter(GLUT-4) was also examined by RT-PCR and in vitro transcription with co-amplification of rat $\beta$-actin gene as an internal standard. Muscular GLUT-4 mRNA expression in SPSGH treated group was increased significantly. These results may suggest that SPSGH lowered blood glucose ascribing to upregulation of muscular GLUT-4 mRNA expression.

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Synthesis and Activity of a Potent ${\alpha}$-glucosidase inhibitor, (1R, 6R, 8S)-cis-1,6-dihydroxypyrrolizidine, and its isomer

  • Jung, Kyeong-Eun;Kang, Yong-Koo;Kim, Dong-Jin;Park, Sang-Woo
    • Archives of Pharmacal Research
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    • v.20 no.4
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    • pp.346-350
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    • 1997
  • The synthesis of cis- and trans-1,6-dihydroxypyrrolizidine starting from trans-4-hydroxy-L-proline and their evaluation as glycosidase inhibitors are reported. The cis-isomer was found to be a potent inhibitor against .alpha.-glucosidase and showed weak inhibitory effect against other glycosidases. The trans-isomer exhibited weak inhibitions of b-glucosidase and amylo-glucosidase and poor inhibition of other glycosidases.

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