• Title/Summary/Keyword: TLC pattern

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Safety Evaluation of Filamentous Fungi Isolated from Industrial Doenjang Koji

  • Lee, Jin Hee;Jo, Eun Hye;Hong, Eun Jin;Kim, Kyung Min;Lee, Inhyung
    • Journal of Microbiology and Biotechnology
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    • v.24 no.10
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    • pp.1397-1404
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    • 2014
  • A few starters have been developed and used for doenjang fermentation but often without safety evaluation. Filamentous fungi were isolated from industrial doenjang koji, and their potential for mycotoxin production was evaluated. Two fungi were isolated; one was more dominantly present (90%). Both greenish (SNU-G) and whitish (SNU-W) fungi showed 97% and 95% internal transcribed spacer sequence identities to Aspergillus oryzae/flavus, respectively. However, the SmaI digestion pattern of their genomic DNA suggested that both belong to A. oryzae. Moreover, both fungi had morphological characteristics similar to that of A. oryzae. SNU-G and SNU-W did not form sclerotia, which is a typical characteristic of A. oryzae. Therefore, both fungi were identified to be A. oryzae. In aflatoxin gene cluster analysis, both fungi had norB-cypA genes similar to that of A. oryzae. Consistent with this, aflatoxins were not detected in SNU-G and SNU-W using ammonia vapor, TLC, and HPLC analyses. Both fungi seemed to have a whole cyclopiazonic acid (CPA) gene cluster based on PCR of the maoA, dmaT, and pks-nrps genes, which are key genes for CPA biosynthesis. However, CPA was not detected in TLC and HPLC analyses. Therefore, both fungi seem to be safe to use as doenjang koji starters and may be suitable fungal candidates for further development of starters for traditional doenjang fermentation.

A Study on Characteristics of the Twin Tree Array Structure of the Triangular Patch Array Antenna (삼각형 패치 트윈 트리 배열 안테나의 특성 연구)

  • Chang, Tae-Soon;Kang, Sang-Won
    • Journal of IKEEE
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    • v.24 no.2
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    • pp.372-377
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    • 2020
  • This paper is based on the triangular inset patch antenna as the basic structure, and the array structure is the twin tree form to improve the antenna's main lobe gain and reduce the side lobe gain. A twin tree structure was implemented by placing two identical trees of 2-4-6 arrays of triangular inset patches side by side. The parametric analysis confirmed that the gap between tree structures arranged side by side is most effective for impedance matching. The fabricated antenna has a gain of 16.74 dBi at 24.15 GHz, and the beam width of the main lobe is 22° in the E-plane and 6° in the H-plane. The antenna size was 125 mm × 50 mm, and a Taconic TLC substrate with a dielectric constant of 3.2 was used. Although the main lobe gain is improved over the twin tree structure, The directivity in the beam pattern due to the mutual interference of the two tree array structure can be improved.

Studies of Morphological Properties and Pyrrolizidine Alkaloids Analysis of Comfrey Cultivating in Korea (국내산 컴프리의 형태학적 특성 및 Pyrrolizidine Alkaloids 분석)

  • 김희연;홍진환;김동술;한상배;이은주;강길진;육창수;박종희;배기환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.6
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    • pp.790-794
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    • 2003
  • This study was carried out to investigate the morphological properties and pyrrolizidine alkaloids of comfrey cultivating in Korea. 12 comfrey samples cultivation in Korea was selected and their appearance (whole plant, leaves, root etc.) were observed by expert and microscopy for morphological analysis. It is confirmed that their species are Symphytum officinale Linnaeus. Samples were extracted by hot MeOH and ultra-sonification. Their extracts contained pyrrolizidine alkaloids, which was identified by TLC analysis. By spraying thin-layer chromatograms of pyrrolizidine alkaloids stable purple spots were developed. But the extracts of chicory, pumpkin and sesame leaves did not show any purple spots. Same HPLC pattern were displayed at about 30 min of retention show peaks an one and the same time.

Fermented Property and Antioxidative Effect of GABA Producing Lactobacillus plantarum from Kimchi (김치 유래 GABA 생성 Lactobacillus plantarum의 발효 및 항상화 특성)

  • Lee, Young-Duck
    • Journal of Food Hygiene and Safety
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    • v.36 no.5
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    • pp.440-446
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    • 2021
  • GABA (γ-aminobutyric acid) has various beneficial effects on human health such as anti-hypertension, diuretic, tranquilizer, sleep induction and anti-stress functions. In this study, the properties and the antioxidizing effects of a fermented solution was investigated by applying GABA producing lactic acid bacteria (LAB) from kimchi to corn silk extract. Lactobacillus plantarum LAB459 was identified by physiological properties, carbohydrate fermentation pattern and 16s rRNA sequence analysis. Also, the GABA production ability of the separated L. plantarum LAB459 was confirmed through TLC and HPLC analysis. Moreover, from the fermentation of corn silk extract with skim milk, it was revealed that approximately 1 ㎍/mg of GABA produced by lyophilized ferments was yielded. Lastly, the flavonoid content and DPPH radical scavenging activity were found to be high in the lyophilized ferments than in the aqueous extracts. Therefore, L. plantarum LAB459 is considered to be used as a starter culture for various fermented foods or in food and medicinal materials.

Production of Tropane Alkaloids by Hairy Root Cultures of Scopolia parviflora (미치광이풀(Scopolia parviflora)의 모상근 배양에 의한 Tropane Alkaloid 생산)

  • 안준철
    • Journal of Plant Biology
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    • v.36 no.3
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    • pp.225-231
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    • 1993
  • Transformed hairy roots of Scopolia parviflora, producing tropane alkaloids and native to Korea, were obtained following infection of rhizome segments with Agrobacterium rhizogenes A4. Each root tip induced from inoculum sites excised and cultured in MS agar or liquid medium. About seventy of hairy root clones were established. Among them, several fast growing hairy root clones were examined for alkaliod content. Two dimensional TLC analysis showed that the tropane alkaloid pattern of hairy root was more complicated than that in the rhizome of mother plant. On the other hand, some hairy root clones did not produce scopolamine and hyoscyamine. In HPLC analysis, some hairy root clones yield higher levels of scopolamine and hyoscyamine than those of mother plant rhizome which used for infection. Scopolamine and hyoscyamine were identified by comparison of their retention times and of their spectra data with those of authentic compounds.

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The improvement of ginsenoside accumulation in Panax ginseng as a result of γ-irradiation

  • Kim, Dong Sub;Song, Mira;Kim, Sun-Hee;Jang, Duk-Soo;Kim, Jin-Baek;Ha, Bo-Keun;Kim, Sang Hoon;Lee, Kyung Jun;Kang, Si-Yong;Jeong, Il Yun
    • Journal of Ginseng Research
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    • v.37 no.3
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    • pp.332-340
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    • 2013
  • In this study, gamma rays were used to irradiate embryogenic calli induced from cotyledon explants of Panax ginseng Meyer. After the embryogenic calli were irradiated, they were transferred to adventitious roots using an induction medium; next, mutated adventitious root (MAR) lines with a high frequency of adventitious root formations were selected. Two MAR lines (MAR 5-2 and MAR 5-9) from the calli treated with 50 Gy of gamma rays were cultured on an $NH_4NO_3$-free Murashige and Skoog medium with indole-3-butyric acid 3 mg/L. The expression of genes related to ginsenoside biosynthesis was analyzed using reverse transcription polymerase chain reaction with RNA prepared from native ginseng (NG), non-irradiated adventitious root (NAR) and 2 MAR lines. The expression of the squalene epoxidase and dammarenediol synthase genes was increased in the MAR 5-2 line, whereas the phytosterol synthase was increased in the MAR 5-9 line. The content and pattern of major ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, and Rg1) were analyzed in the NG, NAR, and 2 MAR lines (MAR 5-2 and MAR 5-9) using TLC and HPLC. In the TLC analysis, the ginsenoside patterns in the NG, NAR, and 2 MAR lines were similar; in contrast, the MAR 5-9 line showed strong bands of primary ginsenosides. In the HPLC analysis, compared with the NG, one new type of ginsenoside was observed in the NAR and 2 MAR lines, and another new type of ginsenoside was observed in the 2 MAR lines irradiated with gamma rays. The ginsenoside content of the MAR 5-9 line was significantly greater in comparison to the NG.

Identification and Growth Activity to Bifidobacterium spp. of Locust Bean Gum Hydrolysates by Trichoderma harzianum ${\beta}$-mannanase (Trichoderma harzianum 유래 ${\beta}$-mannanase에 의한 Locust Bean Gum 가수분해 올리고당의 동정 및 Bifidobacterium spp.에 대한 생육활성)

  • Kim, Yu-Jin;Park, Gwi-Gun
    • Applied Biological Chemistry
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    • v.48 no.4
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    • pp.364-369
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    • 2005
  • This study was performed to elucidate substrate specificity to the locust bean gum galactomannan by Trichoderma harzianum ${\beta}-mannanase$. The medium composition for enzyme production were determined 3% cellulose, 3% corn steep liquor, 1% $KH_2PO_4$, 0.2% $(NH_4){_2}SO_4$, and incubated for 115 hr at $28^{\circ}C$. The ${\beta}-mannanase$ exhibited maximum activity at pH 4.5 and $60^{\circ}C$. Locust bean gum galactomannan was hydrolyzed by the ${\beta}-mannanase$, and then hydrolysates separated by activated carbon column chromatography. The main hydrolysates were composed of D.P 4 and 7 galactosyl mannooligosaccharides by TLC. For the elucidate the structure of D.P 4 and 7 oligosaccharides, methylation analysis was performed. D.P 4 and 7 were identified as M-M-M-M and M-M-M-M-M (G- and M-represent ${\alpha-1,6-D-galactosidic\;and\;{\beta}-1,4-mannosidic$ linkages, respectively). //G-G To investigate the effects of locust bean gum galactosyl mannooligosaccharides on the in vitro growth of B. longum, B. bifidum, B. infantis, and B. breve, Bifidobacterium spp. were cultivated individually on the modified-MRS medium containing carbon source such as D.P 4 and 7 galactosyl mannooligosaccharides, respectively. B. longum grew up 3.4-fold and 4.3-fold more effectively by the replacement of D.P 4 and 7 galactosyl mannooligosaccharides as the carbon source in a comparasion of standard MRS.

Purification of Xylogone sphaerospora ${\beta}$-mannanase and Growth Activity of Bifidobacterium spp. by Konjac Glucomannan Hydrolysates (Xylogone sphaerospora 유래 ${\beta}$-mannanase 정제 및 Konjac Glucomannan 가수분해 올리고당의 중합도별 Bifidobacterium spp.에 대한 증식활성)

  • Lee, Hee-Jung;Park, Gwi-Gun
    • Applied Biological Chemistry
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    • v.51 no.3
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    • pp.159-163
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    • 2008
  • Xylogone sphaerospora ${\beta}$-mannanase was purified by Sephadex G-100 column chromatography. The specific activity of the purified enzyme was 8.44 units/ml protein, representing an 56.27-folds purification of the original crude extract. The final preparation thus obtained showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight was determined to be 42kDa. Konjac glucomannan was hydrolyzed by the purified ${\beta}$-mannanase, and then the hydrolysates was separated by activated carbon column chromatography. The main hydrolysates were composed of D.P. (Degree of Polymerization) 3 and 4 glucomannooligosaccharides. For elucidate the structure of D.P 3 and 4 glucomannooligosaccharides, sequential enzymatic action was performed. D.P 3 and 4 were identified as M-G-M and M-M-G-M (G- and M- represent glucosidic and mannosidic link-ages). To investigate the effects of konjac glucomannooligosaccharides on in vitro growth of Bifido-bacterium longum, B. bifidum, B. infantis, B. adolescentis, B. animalis, B. auglutum and B. breve. Bifidobacterium spp. were cultivated individually on the modified-MRS medium containing carbon source such as D.P. 3 and D.P. 4 glucomannooligosaccharides, respectively. B. longum and B. bifidum grew up 3.9-fold and 2.8-fold more effectively by the treatment of D.P. 4 glucomannooligosaccharides, compared to those of standard MRS medium. Especially, D.P. 4 was more effective than D.P. 3 glucomannooligosaccharide on the growth of Bifidobacterium spp.

Preparation of $Gal^3Man_4(6^3-mono-{\alpha}-D-galacto-pyranosyl-{\beta}-mannotetraose)$ by Bacillus sp. ${\beta}-mannanase$ and Growth Activity to Intestinal Bacteria (Bacillus sp.유래 ${\beta}-mannanase$에 의한 $Gal^3Man_4(6^3-mono-{\alpha}-D-galacto-pyranosyl-{\beta}-mannotetraose)$ 조제 및 장내세균에 대한 생육활성)

  • Kim, Sang-Woo;Park, Gwi-Gun
    • Applied Biological Chemistry
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    • v.47 no.4
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    • pp.379-383
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    • 2004
  • For the elucidation of substrate specificity to the brown copra meal by Bacillus sp. ${\beta}-mannanase.$, the enzymatic hydrolysate after 24 hr of reaction was heated in a boiling water bath for 10 min, and then centrifuged to remove the insoluble materials from hydrolysates. The major hydrolysates composed of D.P 5 and 7 galactosyl mannooligosaccharides. For the separate of galactosyl mannooligosaccharides, the supernatant solution of 150 ml was put on a first activated carbon column. The column was then washed with 5 l of water to remove mannose and salts. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol, at the flow rate of 250 ml per hour. The sugar composition in each fraction tubes was examined by TLC and FACE analysis. The combined fraction from F3 was concentrated to 30 ml by vacuum evaporator. Then put on a second activated carbon column. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol (total volume: 5 l), at the flow rate of 250 ml per hour. The eluent was collected in 8 ml fraction tubes, and the total sugar concentration was measured by method of phenol-sulfuric acid. The major component of F2 separated by 2nd activated carbon column chromatography were identified $Gal^3Man_4(6^3-mono-{\alpha}-D-galactopyranosyl-{\beta}-mannotetraose)$. To investigate the effects of brown copra meal galactomannooligosaccharides on growth of Bifidobacterium longum, B. bifidum were cultivated individually on the modified-MRS medium containing carbon source such as $Gal^3Man_4$, compared to those of standard MRS medium.

Reaction Pattern of Bacillus cereus D-11 Chitosanase on Chitooligosaccharide Alcohols

  • Gao, Xing-Ai;Jung, Woo-Jin;Kuk, Ju-Hee;Park, Ro-Dong
    • Journal of Microbiology and Biotechnology
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    • v.19 no.4
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    • pp.358-361
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    • 2009
  • The purified endochitosanase(Mw 41 kDa) from bacterium Bacillus cereus D-11 hydrolyzed chitooligomers $(GlcN)_{5-7}$ into chitobiose, chitotriose, and chitotetraose as the final products. The minimal size of the oligosaccharides for enzymatic hydrolysis was a pentamer. To further investigate the cleavage pattern of this enzyme, chitooligosaccharide alcohols were prepared as substrates and the end products of hydrolysis were analyzed by TLC and HPLC. The chitosanase split $(GlcN)_4GlcNOH$ into $(GlcN)_3+(GlcN)_1GlcNOH$, and $(GlcN)_5GIcNOH$ into $(GlcN)_4+(GlcN)_1GlcNOH$ and $(GlcN)_3+(GlcN)_2GlcNOH$. The heptamer $(GlcN)_6GlcNOH$ was split into $(GlcN)_5$ [thereafter hydrolyzed again into $(GlcN_3+(GlcN)2]+(GlcN)_1GlcNOH$, $(GlcN)_4+(GlcN)_2GlcNOH$, and $(GlcN)_3+(GlcN)_3GlcNOH$, whereas $(GlcN)_{1-3}GlcNOH$ was not hydrolyzed. The monomers GlcN and GIcNOH were never detected from the enzyme reaction. These results suggest that D-11 chitosanase recognizes three glucosamine residues in the minus position and simultaneously two residues in the plus position from the cleavage point.