• Title/Summary/Keyword: HPLC Pattern

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Purification and Characterization of Bacteriocin J105 Produced by Lactococcus latis subsp. lactis J105 Isolated from Kimchi

  • Kwak, Gyu-Suk;Kim, Sung-Koo;Jun, Hong-Ki
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.275-280
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    • 2001
  • Bacteriocin J105 is a proteinaceous inhibitory substance produced by Latococcus latis subsp. lactis j105 isolated from Kimchi. Bacteriocin J105 was purified to homogeneity by the pH-dependent adsorption-desorption method and reverse-phase HPLC from the culture broth of Lactococcus lactis subsp. lactis J105. Purification of bacteriocin J105 resulted in a 1.47-fold increase in the specific activity and the recovery was 1.5%. Its molecular mass measured by the electrophoretic pattern in the sodium, dodecyl sulfate polyacrylamide gel was about 3.4 kDa. It was stable at $121^{\circ}C$ for 15 min at pH between 2 and 4. However, at pH above 5, bacteriocin was rapidly inactivated. Twenty-one residues from the N-terminal portion of bacteriocin J105 were sequenced using sequence analysis of lantibiotics. Bacteriocin J105 showed significant homology with known nisin A from lactic acid bacteria.

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Isolation and Identification of Tetrodotoxin-producing Marine Microorganism from Pufferfish (복어로부터 복어독(Tetrodotoxin) 생성능이 있는 해양 미생물의 분리 및 동정)

  • 윤성준;차병윤;이명자;정동윤;송병권;김희숙;김동수;이은열
    • Journal of Life Science
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    • v.9 no.6
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    • pp.653-658
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    • 1999
  • A novel marine microorganism, Vibrio sp. YE-101, was isolated from pufferfish and investigated for its ability to synthesize tetrodotoxin (TTX). Various strains isolated from the intestine of pufferfish were grown on TCBS agar plate, and then cultured on Ocean Research Institute (ORI) medium supplemented with 3% NaCl at 23$^{\circ}C$ for 3days. The cells were harvested, disrupted, fractionated by Bio-Gel P-2 column chromatography and then TTX-producing strain, Vibrio sp. YE-101, was identified using mouse bioassay. The isolated TTX from Vibrio sp. YE-101 was also analyzed and identified by HPLC and gas chromatography-mass spectrometer (GC-MS). The mass fragmentation of trimethylsilyl derivatives of C9-base of TTX from Vibrio sp. YE-101 was interpreted and the pattern of fragmentation was same with that of authentic standard. The purfied TTX was also positive to the mouse bioassay, which clearly represents that Vibrio sp. YE-101 can synthesize TTX.

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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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Studies on the Manufacturing of Soft Ginseng Drink Part I. Purification procedure of raw ginseng extracts (인삼청량음료 제조에 관한 연구)

  • 양재원;성현순;박명한;김우정;홍순근
    • Journal of Ginseng Research
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    • v.4 no.1
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    • pp.72-87
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    • 1980
  • Red ginseng tails were extracted with ethanol solutions over a range of concentrations and temperature conditions. Investigations were carried out to study the effects of treatments on yields, soluble solids, saponin and precipitate occured in red ginseng extract beverage during storage. It was found that: (1) Higher concentration of ethanol at low temperature resulted in less yield of crude extract (2) The amount of precipitate in the non-purified extract beverage were less with decrease in ethanol concentration used (3) The treatment for purification of extracts and storage of purified extract at 37$^{\circ}C$ for 6 months had no effect on HPLC chromatogram pattern of saponins (4) The amount of purified extract decreased by purification treatment and more decrease was found as the temperature and concentration of ethanol increased. For Preparation of red ginseng extract beverage, the treatment of extracts with ethanol at low temperature was found to be more effective to minimize precipitation in tile beverage.

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Effect of Extraction on Chemical Composition of Rcd Ginseng Extract (추출 회수에 따른 홍삼 extract의 성분 조성에 관한 연구)

  • 최강주;김만욱;성현순;홍순근
    • Journal of Ginseng Research
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    • v.4 no.1
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    • pp.88-95
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    • 1980
  • Red ginseng was extracted with water and analyzed for yield, saponin, pectin and other chemical composition. It was found that: (1) The total solid content in extract after 6 times of extraction was 46.8%,: including 13.6% of centrifugal residue; (2) 83.7% of total extractable solids and 86% total saponin was extracted after the initial three runs of extraction. (3) No significant changes were observed in HPLC pattern of extracted saponins over a range of extractions; (4) The ratio of centrifugal residue to total solids increased as the number of extractions increased; (5) The ratios of fat, protein, reducing sugar and pectin contents decreased with repeating extraction while those of crude fiber, total sugar and 35% alcohol insoluble residue increased when they were compared with total solids.

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Enhanced Thermal Stability of a Novel Human Thrombopoietin Mutein under the Various Temperature Conditions

  • Kim, Tae-Soo;Ahn, Hye-Kyung;Lim, Seung-Wook;Hong, Yeon-Joo;Chung, Joo-Young;Koh, Yeo-Wook
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.237.2-237.2
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    • 2003
  • DWP40458 is a novel human thrombopoietin mutein with two additional N-linked glycosylation site. The thermal stability of DWP40458 in both solution and lyophilized form was studied in the temperature range of 4-50$^{\circ}C$, compared with recombinant human TPO (rhTPO). When the aggregation or degradation pattern of DWP40458 and rhTPO solution was characterized by using SDS-PAGE, gel permeation chromatography (GPC) and reverse phase HPLC, it was found that thermostability of DWP40458 was significantly different to rhTPO in the temperature at 25, 40, 40, 50$^{\circ}C$. (omitted)

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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.

Quality comparison of musk products using a chromatographic pattern analysis (사향의 크로마토그램 패턴 분석을 통한 품질비교 연구)

  • Cho, Chong Woon;Gao, Dan;Kang, Jong Seong;Jung, Hyo won;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.36 no.4
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    • pp.41-50
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    • 2021
  • Objectives : This study is to check the quality of Moschus products in Korean markets using a chromatographic analysis. Methods : We collected musk products, two genuine products from Russia (A, B), two authentic products from Hong Kong (C) and Korea (D), the Ministry of Food and Drug Safety (MFDS) standard (E), and two fire ants products (E, F). Results : For identification, TLC analysis of 60% ethanol extracts of each product showed that A, B, C, and E have the suitable patterns before color development at 365 nm UV light for the MFDS regulation in Korea. A clear red spot was observed from the E at Rf 0.6 with 365 nm UV light after color development, but this spot was not found in A and B. For the purity test, a distinct violet spot with Rf 0.87 was observed from the A and B in TLC analysis of methanol extracts of each product at white light, however, this spot was not matched with the impurities of E and F. In HPLC-UV pattern analysis, a similar peak pattern was shown in A, B and E, and similar peaks were observed C, although the similarity was weaker than that of A, B, and E. The F and G showed different peak patterns compared with the peak patterns of other samples. Conclusions : Hence, it is considered that the test methods need flexibility in application for identification and purity test depending on the type of sample.

Antioxidant activity and analysis of proantbocyanidins from pine (Pinus densiflora)needles

  • Park, Yong-Soo;Jeon, Min-Hee;Hwang, Hyun-Jung;Park, Mi-Ra;Lee, Sang-Hyeon;Kim, Sung-Gu;Kim, Mi-Hyang
    • Nutrition Research and Practice
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    • v.5 no.4
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    • pp.281-287
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    • 2011
  • In this study, we evaluated the antioxidant activity of pine needle extracts prepared with hot water, ethanol, hexane, hot water-hexane (HWH), and hot water-ethanol (HWE), using the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical method. The hot water extract possessed superior antioxidant activity than the other extracts. We also compared the antioxidant activity of pine needle extracts through ROS inhibition activity in a cellular system using MC3T3 E-1 cells. The hot water extract exhibited the lowest ROS production. The pattern of HPLC analysis of each extract indicated that the hot water extract contained the highest proanthocyanidin level. The pine needle hot-water extract was then isolated and fractionated with Sephadex LH-20 column chromatography to determine the major contributor to its antioxidant activity. The No.7 and 12 fractions had high antioxidant activities, that is, the highest contents of proanthocyanidins and catechins, respectively. These results indicate that the antioxidant activity of procyanidins from the hot water extract of pine needles is positively related to not only polymeric proanthocyanidins but also to monomeric catechins. Moreover, the antioxidant activity of the pine needle hot water extract was similar to well-known antioxidants, such as vitamin C. This suggests that pine needle proanthocyanidins and catechins might be of interest for use as alternative antioxidants.

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.