• Title/Summary/Keyword: cellobiose

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Whole-Cell Biocatalysis for Producing Ginsenoside Rd from Rb1 Using Lactobacillus rhamnosus GG

  • Ku, Seockmo;You, Hyun Ju;Park, Myeong Soo;Ji, Geun Eog
    • Journal of Microbiology and Biotechnology
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    • v.26 no.7
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    • pp.1206-1215
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    • 2016
  • Ginsenosides are the major active ingredients in ginseng used for human therapeutic plant medicines. One of the most well-known probiotic bacteria among the various strains on the functional food market is Lactobacillus rhamnosus GG. Biocatalytic methods using probiotic enzymes for producing deglycosylated ginsenosides such as Rd have a growing significance in the functional food industry. The addition of 2% cellobiose (w/v) to glucose-free de Man-Rogosa-Sharpe broths notably induced β-glucosidase production from L. rhamnosus GG. Enzyme production and activity were optimized at a pH, temperature, and cellobiose concentration of 6.0, 40℃, and 2% (w/v), respectively. Under these controlled conditions, β-glucosidase production in L. rhamnosus GG was enhanced by 25-fold. Additionally, whole-cell homogenates showed the highest β-glucosidase activity when compared with disrupted cell suspensions; the cell disruption step significantly decreased the β-glucosidase activity. Based on the optimized enzyme conditions, whole-cell L. rhamnosus GG was successfully used to convert ginsenoside Rb1 into Rd.

Characterization of β-glucosidase from Brown Rot Fungus, Laetiporus sulphureus

  • Lee, Jae-Won;Park, Jun-Yeong;Gwak, Ki-Seob;Koo, Bon-Wook;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.35 no.5
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    • pp.100-108
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    • 2007
  • $\beta$-Glucosidase from Laetiporus sulphureus among the enzymes related to lignocellulosic biomass degradation to sugars for using alternative bioethanol production was characterized. The highest activity of $\beta$-glucosidase was obtained on cellobiose at shaking culture. For the characterization and purification of $\beta$-glucosidase culture solution was concentrated and then purified by FPLC using ion exchange and size exclusion column. According to the results of SDS-PAGE, native PAGE and microfluidic system of purified enzyme, protein band was observed at about 132 kDa. Optimal pH and temperature of purified $\beta$-glucosi-dase were 5.0 and $60^{\circ}C$, respectively. In the kinetic properties of $\beta$-glucosidase on various substrates such as sophorose, gentiobiose and cellobiose, $K_m$ was 0.81, 1.07 and 1.70 mM, respectively.

β-Glucosidase Formation In Cellulomonas sp. (Cellulomonas sp.의 β-글루코시다아제 생성)

  • Choi, Woo-Young
    • Korean Journal of Agricultural Science
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    • v.3 no.2
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    • pp.225-234
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    • 1976
  • To elucidate ${\beta}$-glucosidase formation on various carbon scurces by cellulolytic bact-eia, Cellulomonas sp. CS1-1, the strain was grown on Nutrient Yeast Broth, carboxymethyl cellulose, avicel and cellobiose using a Ouickfit FVIL fermentor operated in batch, and the growth characteristics on those substrates and ${\beta}$-glucosidase distribution of extra and intracellular enzyme components were studied. The results were: 1) ${\beta}$-glucosidase was always intracellular, and was formed under all growth conditions tested, ii) but levels of relative activities were higher when the culture was grown on cellobiose and on avicel, iii) the relative activities were always maximum during the growth phase of the organism irrespective of the substrate used.

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Penicillium verruculosum으로부터 Cellobiohydrolase의 정제 및 특성

  • 조남철;김강화;전순배;정기철
    • Proceedings of the Microbiological Society of Korea Conference
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    • 1991.04a
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    • pp.187-199
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    • 1991
  • Penicillium verruculosum 배양액으로부터 소단위체 분자량이 60,000(cellobiohydrolase I)과 66,000(cellobiohydrolase II) 및 76,000(cellobiohydrolase III)인 cellobiohydrolase를 분리 정제하여 그들의 일반적 특성을 검토하였다. 이들 세 효소들은 모두 당단백질로서 cellobiohydrolase I, II 그리고 III는 각각 $8.6\%$, $4.2\%$ 그리고 $8.5\%$의 당함량을 나타냈으며 세 효소 모두 pH 4.5 - 5.0, 온도 $50 - 60^{\circ}C$에서 최적조건을 나타냈다. Cellobiohydrolase I, II 및 III는 모두 Avicel, cotton, 여지 등의 결정성 섬유소 뿐만 아니라 carboxymethyl cellulose에도 활성도를 나타냈다. 정제된 cellobiohydrolase I, II 및 III의 Avicel에 대한 비 활성도는 각각 0.07, 0.10, 그리고 0.07 unit per mg. of protein 이었으며 Avicel 분해 생성물은 거의 cellobiose였다. 또한 p-Nitrophenyl-$\beta$-D-cellobioside를 기질로 하였을 때 이들 세 효소 모두 포도당에 의해 활성도가 저해되지 않은 반면 cellobiose에 의해서는 저해되었다. 아미노산조성, 트립신에 의한 펩타이드들의 용출양상 그리고 항체를 이용한 Immunoblotting 결과로부터 cellobiohydrolase II와 III는 동일 유전자산물이거나 1차 구조가 거의 유사한 단백질로 추정된다. Cellobiohydrolase II로부터 분리한 2 개의 펩타이드의 아미노산 서열은 Trichoderma cellobiohydrolase I과 상동성을 보였으며 또한 이 두 호소의 아미노산조성은 매우 유사하였다.

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Studies on the Clostridium bovis sp. nov., the predominant species isolated from the feces of Holstein cattle (홀스타인 젖소의 분변에서 우세균종으로 분리되는 새로운 Clostridium bovis 에 관한 연구)

  • Lee, Wan-kyu
    • Korean Journal of Veterinary Research
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    • v.34 no.1
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    • pp.99-105
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    • 1994
  • Clotridium bovis sp. nov, is described on the basis of 5 strains isolated from the feces of Holstein cattle. The isolate are gram-positive, motile, strict anaerobic spore forming rods. They differ from all the validly described related species of the genus Clostridium in carbohydrate fermentation pattern, G+C mol% and DNA homologies. Acid is produced from arabinose, xylose, glucose, mannose, fructose, galactose, sucrose, maltose, cellobiose, lactose, trehalose, melibiose, raffinose, inulin and salicin. Major end products in PYFG broth are large amounts of butyric acid and lactic acid, and trace amounts of acetic and succinic acids. The G+C mol% of DNA from the type strain is 26 mol%. The type strain of Clostridium bovis is Catt $66^T$ strain.

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Isolation and Identification of Vibrio vulnificus and Vibrio parahaemolyticus from Coast of Pusan and Daechon (부산과 대천 해안에서 Vibrio vulnificus와 Vibrio parahaemolyticus의 분리 및 동정)

  • Ju, Jin-Woo;Park, Min-Jung;Heo, Moon-Soo;Jung, Cho-Rok
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.4
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    • pp.309-316
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    • 2000
  • This study was focused on the isolation of pathogenic Vibrio species, V. vulnificus and V. parahaemolyticus from marine environment from May to July of 1999. Isolation sites were coast near by Pusan and Daechon. The results obtained were as follows: 1. Seventy strains of V. parahaemolyticus and 19 strains of V. vulnificus were isolated from a total of 120 specimens. 2. Nineteen strains of V. vulnificus did not fermented arabinose and salicin but fermented lactose and cellobiose. All of V. parahaemolyticus isolates did not fermented lactose and cellobiose. 47 strains of V. parahaemolyticus fermented arabinose but 53 strains did not fermented salicin. 3. V. vulnificus and V. parahaemolyticus isolates showed three different API index numbers with 5046105 and 4346107 dominant. 4. V. vulnificus did not grow on 0% and 8% NaCl containing medium. V. parahaemolyticus grew on 8% NaCl containing medium. 5. V. vulnificus isolates and V. parahaemolyticus revealed different outer membrane protein profiles on SDS-PAGE.

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Characteristics of Cellulomonas fimi $\beta$-glucosidase expressed in Escherichia coli (대장균에서 발현되는 Cellulomonas fimi $\beta$-glucosidase의 효소학적 특징)

  • Kim, Ha-Kun
    • The Journal of Natural Sciences
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    • v.8 no.2
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    • pp.57-61
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    • 1996
  • The $\beta$-glucosidase enzyme was purified from E. coli carrying Cellulomonas fimi $\beta$-glucosidase gene. SDS-PAGE and analytical gel filtration revealed that molecular weight of this enzyme was 56,000 dalton and consisted of a single polypeptide.Inhibition caused by heavy metals and activation by dithiothreitol suggest the existence of essential thiol group in the enzyme. The enzyme was not active on maltose (glucose $\alpha$-1,4-glucose) which has a $\alpha$-linkage, whereas it was active on lactose (glucose $\beta$-1,4-glucose), PNPG (p-nitrophenyl $\beta$-D-glucopyranoside) and PNPC (p-nitrophenyl $\beta$-D-cellobioside), although its reaction rates were different.

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Ethanol Production from Various Sugars and Cellulosic Biomass by White Rot Fungus Lenzites betulinus

  • Im, Kyung Hoan;Nguyen, Trung Kien;Choi, Jaehyuk;Lee, Tae Soo
    • Mycobiology
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    • v.44 no.1
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    • pp.48-53
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    • 2016
  • Lenzites betulinus, known as gilled polypore belongs to Basidiomycota was isolated from fruiting body on broadleaf dead trees. It was found that the mycelia of white rot fungus Lenzites betulinus IUM 5468 produced ethanol from various sugars, including glucose, mannose, galactose, and cellobiose with a yield of 0.38, 0.26, 0.07, and 0.26 g of ethanol per gram of sugar consumed, respectively. This fungus relatively exhibited a good ethanol production from xylose at 0.26 g of ethanol per gram of sugar consumed. However, the ethanol conversion rate of arabinose was relatively low (at 0.07 g of ethanol per gram sugar). L. betulinus was capable of producing ethanol directly from rice straw and corn stalks at 0.22 g and 0.16 g of ethanol per gram of substrates, respectively, when this fungus was cultured in a basal medium containing 20 g/L rice straw or corn stalks. These results indicate that L. betulinus can produce ethanol efficiently from glucose, mannose, and cellobiose and produce ethanol very poorly from galactose and arabinose. Therefore, it is suggested that this fungus can ferment ethanol from various sugars and hydrolyze cellulosic materials to sugars and convert them to ethanol simultaneously.

Enzymatic Synthesis of $\beta$ -Glucosides of Aromatic Alcohols and Monoterpene Alcohols (효소적 방법에 의한 방향족 알코올류 및 Monoterpene Alcohol류의 배당체 합성)

  • ;;;;Suzuki Yukio
    • Journal of the Korean Society of Tobacco Science
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    • v.25 no.1
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    • pp.70-79
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    • 2003
  • $\beta$ -Glucosidase-catalysed synthesis of glucosides with aromatic alcohols and monoterpene alcohols as accepters and cellobiose as a donor in the presence of various commercial $\beta$ -glucosidases were described. $\beta$ -Glucosidases from Aspergillus niger spp,. Trichoderma spp., Penicillium sup. and bitter almond have been shown to catalyze synthesis of $\beta$ -glucosides of benzyl alcohol, 2-hydroxybenzyl alcohol, 4-hydroxybenzyl alcohol, 2-phenylethyl alcohol, geraniol and citronellol in the presence of cellobiose as sugar donor. Among enzyme preparations tested, each $\beta$ -glucosides prepared from Aspergillus niger were isolated in the pure state by Diaion HP-20 and silica gel column chromatography. The products were identified as $\beta$ -glucosyl products of benzyl alcohol, 2-hydroxyhenzyl alcohol, 4-hydroxybenzyl alcohol, 2-phenyl ethyl alcohol, geraniol and citronellol by spectrometry (UV, IR, $^1$H-NMR, $^{13}$ C-NMR) and enzymatic hydrolysis with $\beta$ - glucosidase. Monoterpene alcohols with a sterically hindered hydroxyl group, such as linalool, $\ell$-menthol and $\alpha$-terpineol were not used as acceptors in transglycosylation reaction.

Factors Influencing Biohydrogenation and Conjugated Linoleic Acid Production by Mixed Rumen Fungi

  • Nam, In-Sik;Garnsworthy, Philip C.
    • Journal of Microbiology
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    • v.45 no.3
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    • pp.199-204
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    • 2007
  • The objective of this study was to evaluate the effect of soluble carbohydrates (glucose, cellobiose), pH (6.0, 6.5, 7.0), and rumen microbial growth factors (VFA, vitamins) on biohydrogenation of linoleic acid (LA) by mixed rumen fungi. Addition of glucose or cellobiose to culture media slowed the rate of biohydrogenation; only 35-40% of LA was converted to conjugated linoleic acid (CLA) or vaccenic acid (VA) within 24 h of incubation, whereas in the control treatment, 100% of LA was converted within 24 h. Addition of VFA or vitamins did not affect biohydrogenation activity or CLA production. Culturing rumen fungi at pH 6.0 slowed biohydrogenation compared with pH 6.5 or 7.0. CLA production was reduced by pH 6.0 compared with control (pH 6.5), but was higher with pH 7.0. Biohydrogenation of LA to VA was complete within 72 h at pH 6.0, 24 h at pH 6.5, and 48 h at pH 7.0. It is concluded that optimum conditions for biohydrogenation of LA and for CLA production by rumen fungi were provided without addition of soluble carbohydrates, VFA or vitamins to the culture medium; optimum pH was 6.5 for biohydrogenation and 7.0 for CLA production.