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

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Isolation, Identification and Mutant Development of Butanol Tolerance Bacterium (부탄올 내성 미생물의 분리, 동정 및 변이주의 개발)

  • Jung, Hyesook;Lee, Jinho
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.26-32
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    • 2013
  • Butanol-resistant bacteria were isolated from butanol solvent. The cell growth of isolated strains declined with increasing concentrations of butanol, and isolated strain BRS02 displayed more resistance to 12.5 g/L of butanol than other isolated strains. In addition, strain BRS251, which was resistant to even higher concentrations of butanol, was developed by the mutation of BRS02 using UV. BRS251 could grow in LB medium containing up to 17.5 g/L of butanol, 32.5 g/L of propanol, or 6 g/L of pentanol, whereas the control strain Escherichia coli was found to be tolerant to 7.5 g/L of butanol, 20 g/L of propanol, or 2 g/L of pentanol. The isolated BRS02, a Gram(+) bacterium seen to have a cocci form under the microscope, grew in 6.5% NaCl. According to biochemical tests, BRS02 can metabolize and produce acid with D-galactose, D-maltose, D-mannitol, D-mannose, methyl-${\beta}$-Dglucopyranoside, D-ribose, sucrose, or D-trehalose, as carbon sources. Also, this strain showed resistance to bacitracin, vibriostatic agent O/129, and optochin, alongside positive activities for arginine dihydrolase, ${\alpha}$-glucosidase, and urease. The BRS02 strain was identified as Staphylococcus sp. by analyses of the 16S rRNA gene, phylogenetic tree, and biochemical tests.

Conversion of Ginsenoside Rd to Compound K by Crude Enzymes Extracted from Lactobacillus brevis LH8 (Lactobacillus brevis LH8이 생산하는 효소에 의한 Ginsenoside Rd의 Compound K로의 전환)

  • Quan, Lin-Hu;Liang, Zhiqi;Kim, Ho-Bin;Kim, Se-Hwa;Kim, Se-Young;Noh, Yeong-Deok;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.32 no.3
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    • pp.226-231
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    • 2008
  • Ginsenosides have been regarded as the principal components responsible for the pharmacological and biological activities of ginseng. Absorption of major ginsenosides from the gastrointestinal tract is extremely low, when ginseng is orally administered. In order to improve absorption and its bioavailability, conversion of major ginsenosides into more active minor ginsenoside is very much required. Here, we isolated lactic acid bacterium (Lactobacillus brevis LH8) having ${\beta}-glucosidase$ activity from Kimchi. Bioconversion ginsenoside Rd by this bacterium in different temperatures was investigated. The maximum activities of crude enzymes precipitated by ethanol were shown in $30^{\circ}C$ and then gradually decreased. In order to compare the effect of pH, the crude enzymes of L. brevis LH8 were mixed in 20mM sodium phosphate buffer (pH 3.5 to pH 8.0) and reacted ginsenoside Rd. Ginsenoside Rd was almost hydrolyzed between pH 6.0 and pH 12.0, but not hydrolyzed under pH 5.0 and above pH 13.0. Ginsenoside Rd was hydrolyzed after 48 h incubation, whereas ginsenoside F2 appeared from 48 h to 72 h, and ginsenoside Rd was almost converted into compound K after 72 h.

Assessment of the Ability of Extracellular Enzyme Production in Hybrid Strains of Lentinula edodes by Chromogenic Reaction-based Plate Assay (발색반응 분석법을 이용한 표고 교배균주의 세포외효소 분비 능력 평가)

  • Kwon, Hyuk-Woo;Kim, Jun-Young;Ko, Han-Gyu;Park, Heung-Soo;Kim, Seong-Hwan
    • The Korean Journal of Mycology
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    • v.39 no.2
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    • pp.99-104
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    • 2011
  • Shiitake breeding requires the procedures of mating of two different parental strains and selection of hybrid strains that have good traits for the mushroom production. In this study, we tested the possibility of the use of chromogenic plate-based assay for extracellular enzyme production in order to assess and find good biochemical properties-possessed hybrid strains that were generated from genetic cross of the monokaryotic strains derived from two different parental strains of Lentinula edodes Sanjo-101ho and Sanjo-108ho. We observed that there was difference in the ability of producing ${\beta}$-glucosidase, avicelase, CM-cellulase, amylase, pectinase, xylanase, and protease among the monokaryotic strains. We could also comparatively assess that the ability of the seven extracellular enzymes production in the hybrid strains depended on the mating combination of the monokaryotic strains. Our results demonstrate that the assessment method for extracellular enzyme production using chromogenic plate assay could be usefully applied to the assessment of the hybrid strains derived from the breeding procedure of L. edodes.

Cellulose Degradation and Extracellulat Enzymatic Activity of the Mud Flat in Sunchon Bay (순천만 갯벌 토양의 섬유소 분해능 및 체외효소 활성)

  • 백근식;최지혁;성치남
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.130-135
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    • 2000
  • Decomposition rate of organic matiter in the mud flat of Sunchon Bay was estimated. Physicochemical parameters, cellulose degradation rate. distribution of heterotrophic bacteria, and extracellular enzymatic activities were measured from August 1997 to July 1998. Soil temperatures, water contents, concentration of $PO_4$-P and organic matter were -1-~$30^{\circ}C$, 42.1-53.1%, 0.0779-0.1961 mgig and 1.99-7.64%, respectively. Decomposition rate of cellulose film ranged from 7.7 to 100%imonth, high in summer and low in winter. The number of heterotrophic bacteria ranged from $0.87{\times}10^6 to 3.6{\times}10^7 $CUFsIg dq soil. Enzymatic activities of phosphatase, $\alpha$-D-gluEosidase, $\beta$-D-glucosidase and cellobiohydrolase, which were measured as decomposition rate of methylumbelliferyl(MLiF)-substrate, were 152.23-1779.80 nMIhr, 2.67-202.18 nM/hr, 5.03-258.26 M h r and 3.42-63.07 nM/hr, respectively Cellulose degradaaon rate and extracellular extracellular enzymatic activities were conelated with each other, and showed high correlation coefticiency with soil temperature.

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Physicochemical Characteristics for the Transformation of Blue Pigments from Genipin of Gardenia jasminoides with Amino Acids (치자 Genipin과 아미노산의 청색소변환반응에 관한 물리화학적 연구)

  • Lee, Jae-Youn;Hahn, Tae-Ryong;Paik, Young-Sook
    • Applied Biological Chemistry
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    • v.41 no.5
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    • pp.399-404
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    • 1998
  • Genipin was obtained from hydrolysis of geniposide isolated from gardenia fruits with ${\beta}-glucosidase$. Reaction of genipin with glycine, alanine, histidine, lysine, phenylalanine and glutamate in aqueous buffer solution converted colorless starting materials to blue pigments. Effect of pH for the formation of blue pigments was tested using UV/Vis spectrophotometer. The optimum pH for the formation of blue pigments was 7.0. No pigment and trace amounts were formed at acidic (pH 3.0) and alkaline (pH 12.0) conditions, respectively. The amount and tincture of blue color were distinct with different amino acids. In contrast with lysine $({\lambda}_{max}=573\;nm)$, glycine $({\lambda}_{max}=595\;nm)$, phenylalanine $({\lambda}_{max}=602\;nm)$ and alanine $({\lambda}_{max}=595\;nm)$, the reaction of genipin with histidine $({\lambda}_{max}=601\;nm)$ and glutamate $({\lambda}_{max}=601\;nm)$ produced relatively small amounts of blue pigments. Rate constants for the formation of blue pigments from genipin with amino acids at various temperatures $(60,\;70,\;80,\;90^{\circ}C,\;pH\;7.0\;phosphate\;buffer)$ were obtained. Rate constants of genipin with basic amino acids were larger than neutral or acidic amino acids. Arrhenius activation energies of the formation of blue pigments indicated that activation energy of glycine $(E_A=9.8\;kcal/mol)$ was especially lower than those of other amino acids $(E_A=13.3{\sim}15.4\;kcal/mol)$.

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Beneficial Effect of the Combination of Oral Administration and Herbal -Acupuncture Stimulation with Several Herb-combind Prescription on Streptozotocin-Induced Diabetic Rats (한약복합처방(韓藥複合處方) 약침(藥鍼) 및 경구투여(經口投與)가 Streptozotoin에 의한 흰쥐의 당뇨병(糖尿病)과 항산화능(抗酸化能)에 미치는 영향)

  • Park, Sa-hyun;Cho, Su-in;Chae, Woo-seok;Cho, Myung-rae
    • Journal of Acupuncture Research
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    • v.22 no.1
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    • pp.1-11
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    • 2005
  • Objective : The present study was carried out to investigate the preventive effect of Several Herb-combind Prescription(SHP) on Streptozotocin (STZ) -induced Diabetes mellitus. Methods : SHP was given to rats with the combination of oral administration and herbal-acupuncture stimulation. The experimental animals were divided into 3 groups : normal group of rats, control group of STZ-induced diabetic rats, sample group with SHP treatment. In vitro test of SHP showed ${\alpha}$-glucosidase inhibition, DPPH radical scavenging activity and inhibition of lipid peroxidation. Experimental diabetes was induced by the injection of STZ(60mg/kg) to the rat via the peritoneum. The effect of SHP on STZ-induced diabetes was observed by measuring the seum level of insulin, glucose, triglyceride, total cholesterol and lipid peroxides. Hepatic activities of catalase and reduced glutathione were examined and insulin granule was observed by immunohistochemical examination. Result : STZ caused hyperglycemia and hypoinsulinemia by a selectively destroying pancreatic ${\beta}$-cell. SHP treatment protected them from the hyperglycemia and hypoinsulinemia. STZ induced increase of serum triglyceride lowered by SHP treatment. And by SHP treatment, pancrease showed a big area with positive immuno-reactivity for presence of insulin with many insulin granules distributed in the ${\beta}$-cells in the islets of Langerhans. Contusions : The SHP treatment showed protective effect on diabetic rat model, and action mechanism of the effect was thought to be concerned with anti-oxidative stress.

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Biogeochemical Reactions in Hyporheic Zone as an Ecological Hotspot in Natural Streams (자연 하천의 생태학적 중요 지점으로서 지표수-지하수 혼합대의 생지화학적 기작)

  • Kim, Young-Joo;Kang, Ho-Jeong
    • Journal of Wetlands Research
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    • v.11 no.1
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    • pp.123-130
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    • 2009
  • Hyporheic zone is an area where hydraulic exchanges occur between surface water and ground water. Such transient area is anticipated to facilitate diverse biogeochemical reactions by providing habitats for various microorganism. However, only a few data are available about microbial properties in hyporheic zone, which would be important in better understanding of biogeochemical reactions in whole streams. The study site is Naesung stream, located in the north Kyoung-Sang Province, of which sediment is sandy with little anthropogenic impacts. Soil samples were collected from a transect placed perpendicular to stream flow. The transect includes upland fringe area dominated by Phragmites japonica, bare soil, and soil adjacent to water. In addition, soil samples were also collected from downwelling and upwelling areas in hyporheic zone within the main channel. Soils were collected from 3 depth in each area, and water content, pH, and DOC were measured. Various microbial properties including extracellular enzyme activities ($\beta$-glucosidase, N-acetylglucosaminidase, phosphatase and arylsulfatase), and microbial community structure using T-RFLP were also determined. The results exhibited a positive correlation between water content and DOC, and between extracellular enzyme activities and DOC. Distinctive patterns were observed in soils adjacent to water and hyporheic zone compared with other soils. Overall results of study provided basic information about microbial properties of hyporheic zone, which appeared to be discernable from other locations in the stream corridor.

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Antioxidant and Antiobesity Activity of Natural Color Resources (천연색소 소재의 항산화 및 항비만 활성)

  • Hwang, Cho-Rong;Tak, Hyun-Min;Kang, Min-Jung;Suh, Hwa-Jin;Kwon, Oh-Oun;Shin, Jung-Hye
    • Journal of Life Science
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    • v.24 no.6
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    • pp.633-641
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    • 2014
  • This study investigated the antioxidant and antiobesity activity of extract powders from the following natural color resources: Polygonum indigo, Black locust, Cochineal, Catechu, Grape, Tesu flower, Henna, Chrysanthemum, Sandalwood Red, Himalayan Rhubarb, and Madder. Total phenol content was the highest in Catechu extract, at 348.25 mg/g. DPPH, ABTS radical scavenging activity and ferric reducing antioxidant power (FRAP) were also higher in Catechu extract. Bleaching inhibition activity in the ${\beta}$-carotene linoleic acid system was the highest in Black locust extract, as was ${\alpha}$-Glucosidase inhibition activity. ${\alpha}$-Amylase inhibition activity was the highest in Catechu extract. Trypsin inhibition activity of Black locust extract was greater than 60%, and ${\alpha}$- chymotrypsin inhibition activity of Catechu extract was greater than 40%. Lipase inhibition activity was the highest Black locust extract, at 52.73%. Viability of 3T3-L1 cells was not affected by treatment with extracts at concentrations of $1.25{\sim}25{\mu}g/ml$. Lipid accumulation in the 3T3-L1 cells was the lowest following treatment with Catechu extract, at 55.8%, and this extract also inhibited adipocyte differentiation. These results suggest that the Catechu and Black locust extracts have high antioxidant and antiobesity activities and can be useful ingredients in functional foods.

Identification and Characterization of Paraconiothyrium brasiliense from Garden Plant Pachysandra terminalis (가든식물 수호초(Pachysandra terminalis)로부터 Paraconiothyrium brasiliense의 분리 및 동정)

  • Choi, Min Ah;Park, Seung Jun;Ahn, Geum Ran;Kim, Seong Hwan
    • The Korean Journal of Mycology
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    • v.42 no.4
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    • pp.262-268
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    • 2014
  • A fungal isolate DUCC5000 from a garden plant Pachysandra terminalis was identified as Paraconiothyrium brasiliense based on the results of morphological and molecular studies. The fungus formed brown to black conidiomata of (0.2-0.7)-2(-3.5) mm singly or as a group on PDA. Conidia measured $2-5{\times}1.8-3{\mu}m$ in size, hyaline, ellipsoid to short-cylindrical, and rounded at both ends. The internal transcribed spacer (ITS) DNA of the isolate shared 100% nucleotide sequence homology with those of known P. brasiliense isolates. Phylogenetic tree inferred from the ITS sequence analysis showed that the DUCC5000 isolate formed a clade with known isolates of P. brasiliense. The fungal mycelia grew better on oatmeal agar than on MEA and PDA. On PDA media under various pH conditions, fungal mycelial growth was observed at pH 9. Colony morphology of the fungus tended to alter depending on the kinds of nutrient media and pH condition. On chromagenic media, the fungus demonstrated its ability to produce extracellular enzymes including amyalse, avicelase, ${\beta}$-glucosidase, protease, and xylanase. However, in pathogenicity testing, no disease symptoms were observed on the leaves of P. terminalis. This strain is the first report on P. terminalis in Korea.

Isolation of Stenotrophomonas rhizopilae Strain GFC09 with Ginsenoside Converting Activity and Anti-wrinkle Effects of Converted Ginsenosides (사포닌 전환 활성 Stenotrophomonas rhizopilae Strain GFC09 균주의 분리 동정 및 전환 사포닌의 주름 개선 효과)

  • Min, Jin Woo;Kim, Hye-Jin;Joo, Kwang-Sik;Kang, Hee-Cheol
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.4
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    • pp.375-382
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    • 2015
  • Ginsenosides (ginseng saponin) as the one of important pharmaceutical compounds of ginseng and is responsible for the pharmacological and biological activities. These ginsenoside produces diverse small molecules ginsenoside which have more pharmacological activities including anti-wrinkle, anti-cancer and anti-oxidant effects. In the present study, we isolated bacteria using esculin agar, to produce ${\beta}$-glucosidase, and we focused on the bio-transformation of ginsenoside. Phylogenetic tree analysis was performed by comparing the 16S rRNA sequences; we identified the strain as Stenotrophomonas rhizopilae strain GFC09. In order to determine the optimal conditions for enzyme activity, the crude enzyme was incubated with 1 mM ginsenoside $Rb_1$. Bioconversion of ginsenoside $Rb_1$ were analyzed using TLC and HPLC. The crude enzyme hydrolyzed the ginsenoside $Rb_1$ along the following pathway: LB: $Rb_1{\rightarrow}Rd{\rightarrow}F_2$ into compound K, TSB: $Rb_1{\rightarrow}Rd{\rightarrow}F_2$. The structure of the hydrolyzed metabolites were identified by NMR. The activity screening tests showed that the conversion product induced the production of type I procollagen in a dose-dependent manner. These results suggested that hydrolyzed ginseng product containing the ginsenoside $F_2$ and compound K could be useful as an active ingredient for wrinkle-care cosmetics.