• Title/Summary/Keyword: H-NMR

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Characteristics and functional analysis of Bacillus strains from the fermented soybean products, Cheonggukjang (전통 발효 청국장으로부터 분리한 Bacillus 균주들의 특성 및 기능 분석)

  • Moon, Ji-Young;Kwon, Soon-Wo;Hong, Seung-Beom;Seok, Soon-Ja;Kim, Jeong-Seon;Kim, Soo-Jin
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.300-307
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    • 2015
  • For selecting Bacillus strains producing high-quality Cheonggukjang, 8 strains were isolated from the different Cheonggukjang samples. Seven of them exhibited the highest 16S rRNA gene sequence similarity value of over 99.9% to Bacillus subtilis subsp. subtilis and one of them showed the similarity to B. licheniformis. All the strains showed positive activities for amylase, cellulase, protease and lipase, and 6 strains are positive for fibrinolytic activity. To confirm the safety of the strains isolated from the samples of Cheonggukjang which are manufactured by traditional method, strains were analyzed for the presence of seven toxin genes of Bacillus cereus and results were found negative. And 7 strains did not produce at all or merely produce both histamine and tyramine, the representative biogenic amines. Biogenic amine degradation analysis by HPLC revealed that, most of them exhibited tyramine degradation activity. For Cheonggukjang fermented by artificial inoculation of selected strains, fermentation property, sensory test, volatile basic nitrogen production and metabolic profiles by $^1H-NMR$ were tested. Seven strains were confirmed to make high-quality Cheonggukjang.

Preparation and Characterization of O-Carboxymethyl Chitosan Ion-complexed Poly(L-Lysine) for Drug and Gene Delivery System (약물 및 유전자 전달체로 응용하기 위한 Poly(L-Lysine)이 결합된 O-Carboxymethyl Chitosan PEG의 제조와 특성)

  • Nam, Joung-Pyo;Kim, Young-Min;Park, Jin-Su;Lee, Eung-Jae;Choi, Chang-Yong;Jang, Mi-Kyeong;Nah, Jae-Woon
    • Applied Chemistry for Engineering
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    • v.21 no.6
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    • pp.643-647
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    • 2010
  • O-carboxymethyl water-soluble chitosan (OCMCh) prepared for enhance the application of chitosan was modified with mthoxy polyethyleneglycol (mPEG) by ion-complex for long circulation in the blood. OCMCh-PEG-PLLs was prepared by forming ion-complex with OCMCh-PEG and Poly(L-Lysine) (PLL) for drug and gene delivery system. The physicochemcal characterisitcs of OCMCh-PEG-PLLs were investigated by FT-IR, $^1H$-NMR. These results showed that CMCh-PEG-PLLs were successfully syntehsized by ion-complex. Particle size distribution and zeta potential of the OCMCh-PEG-PLLs were determined using dynamic light scattering technique. Transmission electron microscopy (TEM) was also used to observe the morphology of the OCMCh-PEG-PLLs. OCMCh-PEG-PLLs have spherical shapes with particle size 290∼390 nm. OCMCh-PEG-PLLs were showed when the feeding amount of mPEG ratio was increased, particle size and zeta potential were decreased. Based on these results, it is possible to introduction of the OCMCh-PEG-PLLs into various biomedical fields such as drug and gene delivery system.

Biosurfactant as a microbial pesticide

  • Lee, Baek-Seok;Choi, Sung-Won;Choi, Ki-Hyun;Lee, Jae-Ho;Kim, Eun-Ki
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.40-44
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    • 2003
  • Soil-borne infectious disease including Pythium aphanidermatum and Rhizoctonia solani causes severe damage to plants, such as cucumber. This soil-borne infectious disease was not controlled effectively by chemical pesticide. Since these diseases spread through the soil, chemical agents are usually ineffective. Instead, biological control, including antagonistic microbe can be used as a preferred control method. An efficient method was developed to select an antagonistic strain to be used as a biological control agent strain. In this new method, surface tension reduction potential of an isolate was included in the ‘decision factor’ in addition to the other factors, such as growth rate, and pathogen inhibition rate. Considering these 3 decision factors by a statistical method, an isolate from soil was selected and was identified as Bacillus sp. GB16. In the pot test, this strain showed the best performance among the isolated strains. The lowest disease incidence rate and fastest seed growth was observed when Bacillus sp. GB16 was used. Therefore this strain was considered as plant growth promoting rhizobacteria (PGPR). The action of surface tension reducing component was deduced as the enhancement of wetting, spreading, and residing of antagonistic strain in the rhizosphere. This result showed that new selection method was significantly effective in selecting the best antagonistic strain for biological control of soil-borne infectious plant pathogen. The antifungal substances against P. aphanidermatum and R. solani were partially purified from the culture filtrates of Bacillus sp. GB16. In this study, lipopeptide possessing antifungal activity was isolated from Bacillus sp. GB16 cultures by various purification procedures and was identified as a surfactin-like lipopeptide based on the Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), high performance liquid chromatography mass spectroscopy (HPLC-MS), and quadrupole time-of-flight (Q-TOF) ESI-MS/MS data. The lipopeptide, named GB16-BS, completely inhibited the growth of Pythium aphanidermatum, Rhizoctonia solani, Penicillium sp., and Botrytis cineria at concentrations of 10 and 50 mg/L, respectively. A novel method to prevent the foaming and to provide oxygen was developed. During the production of surface active agent, such as lipopeptide (surfactin), large amount of foam was produced by aeration. This resulted in the carryover of cells to the outside of the fermentor, which leads to the significant loss of cells. Instead of using cell-toxic antifoaming agents, low amount of hydrogen peroxide was added. Catalase produced by cells converted hydrogen peroxide into oxygen and water. Also addition of corn oil as an oxygen vector as well as antifoaming agent was attempted. In addition, Ca-stearate, a metal soap, was added to enhance the antifoam activity of com oil. These methods could prevent the foaming significantly and maintained high dissolved oxygen in spite of lower aeration and agitation. Using these methods, high cell density, could be achieved with increased lipopeptide productivity. In conclusion to produce an effective biological control agent for soil-borne infectious disease, following strategies were attempted i) effective screening of antagonist by including surface tension as an important decision factor ii) identification of antifungal compound produced from the isolated strain iii) novel oxygenation by $H_2O_2-catalase$ with vegetable oil for antifungal lipopeptide production.

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Antioxidative Activities and Components of Gardenia jasminoides (치자의 항산화 활성 및 활성성분의 분리)

  • Yang, Hye-Jung;Park, Mi-Jung;Lee, Heum-Sook
    • Korean Journal of Food Science and Technology
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    • v.43 no.1
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    • pp.51-57
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    • 2011
  • From the total methanolic extract of Gardenia jasminoides (Rubiaceae), various antioxidative characteristics were identified in terms of nitrite scavenging ability, 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical cation inhibition, superoxide dismutase (SOD)-like activity, and elongation effect of lipid peroxidation using Rancimat. After successive partitioning with n-hexane, chloroform, n-butanol, and water, potent nitrite scavenging abilities were shown in the n-butanol fraction and water fraction, and $IC_{50}$ values were 183 ppm and 194 ppm, respectively. As for ABTS radical cation inhibition, the chloroform fraction was most potent and its $IC_{50}$ was 159 ppm. SOD-like activity was slightly low in all of the fractions. The elongation effect of lipid peroxidation also increased dose-dependently and the antioxidative index (AI) of the total methanolic extract was 2.93 in 1000 ppm, which was more effective than 1.66 of butylated hydroxy anisol in the same concentration. The compounds I and II were isolated through silica gel column chromatography of the active fractions, and identified as geniposide and crocin, respectively, by $^1H-NMR$ spectral data. The $IC_{50}$ values for the nitrite scavenging abilities of geniposide and crocin were 940 ppm and 77 ppm, respectively. In ABTS radical cation inhibition, the $IC_{50}$ values of geniposide and crocin were 684 ppm and 549 ppm, respectively. And the $EC_{50}$ value for SOD-like activity of crocin was 259 ppm, which was much smaller than 453 ppm by the positive control, chlorogenic acid. The $EC_{50}$ value of geniposide could not be identified.

유산균의 항암효과

  • 배형석
    • Proceedings of the Korean Journal of Food and Nutrition Conference
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    • 1997.06b
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    • pp.15-17
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    • 1997
  • 암 발생의 원인 중 80% 이상이 식사습관과 환경오염에 있는 것으로 인정되고 있다. 음식물, 담배, 술, 대기오염, 스트레스, 자외선이 그 대표적인 원인물질로 꼽을 수 있으며 그 중에서도 매일 섭취하는 음식물이 가장 중요한 발암요인으로 지적되고 있다. 대장암과 유방암의 발생에 대한 Wynder 등의 역학조사에서도 식사습관이 암 발생에 중요함을 시사 하고 있다. 동물성 단백질과 지방의 다량 섭취가 대장암과 유방암의 발생을 증가 시키고 섬유질이 풍부한 곡류와 야채의 섭취는 대장암 발생을 억제한다는 상관관계가 밝혀졌다. 그러나 우리가 늘 섭취하는 음식물 자체는 대장암과 유방암을 유발하는 기능이 거의 없다. 섭취된 음식물이 암을 일으키려면 장내 부패 미생물의 분해작용에 의하여 발암물질로 변환되는 과정이 필요하다. 그 발암물질들이 장관으로 흡수 자극함으로써 암을 유발할 수 있다. 반대로 일부장내 미생물들은 장내 발암물질들을 무독화 하거나 숙주의 면역기능을 증강 시킴으로써 암 발생을 억제할 수도 있다는 사실을 간과해서는 안 될 것이다. Mitsuoka는 장내 미생물이 암을 유발하는 중요한 요인이라고 강조하였다. Veer 등은 유산균 발효유를 많이 섭취하면 유방암 발생이 억제됨을, 국제암연구위원회는 섬유질을 많이 섭취하고 있는 핀란드 쿠피오의 거주자들에게는 덴마크의 코펜하겐에 거주하는 사람들에 비하여 대장암 발생율이 1/4에 불과하고 분변내 유산간균수는 100배 높은 사실을 역학조사에서 밝혔다. 이 외에 유산균과 발효유제품의 항암효과에 대한 실험결과들이 많이 발표되었다. 여기에서는 유산균의 항암효과에 대한 지금까지의 관련 자료들을 요약, 정리하여 고찰하고자 한다.높은 당 함량을 나타냈으며, T-AS는 70.3%의 당과 7.8%의 단백질로 구성 되었다. GLG 대부분의 분획들은 60~93%의 glucose로 구성된 다당류 이었으며, 주로 $\beta$-glucose로 구성된 다당류 이었다. 아미노산은 Asp 및 Glu의 산성 아미노산과 Ala, Leu 등의 함량이 높게 나타났으며, 비알칼리 추출물에서 Ser과 Thr의 함량이 높게 나타났다. 다당류 T-AS는 평균 분자량이 2,000 kD와 12kD에서 주 peak를 나타냈으며, 수용성 분획의 평균 분자량은 12kD이고 비수용성 분획은 36~2,000 kD의 평균 분자량 분포를 갖는 것으로 나타났다. IR과 NMR 분석 결과 890 cm-1에서 흡수 peak를 나타내어 $\beta$-(1,3)0glucan과 $\beta$-(1,6)-glucan의 구조를 갖는 다당류로 확인 되었다. T-AS 분획은 C:H:O:N의 함량비가 38.9:5.7:49.6:1.84%이며, 이 물질의 융점은 163 $^{\circ}C$로 연한 갈색을 나타낸다. 분리된 GLG의 항암활성 기전 규명을 위해, in vivo 항암실험, 항보체 활성능, 항체 생성능, serum protein 분비능, 대식세포의 탐식능과 활성능 및 세포간 물질 분비 등의 상관관계를 조사하였다. 다당류 GLG 분획물들 가운데 항보체의 활성이 높았던 분획은 sarcome 180에 대한 항암 활성이 높게 나타났다. 다당류 T-AS의 보체 활성화 기작은 classical과 alternative complement pathway의 양 경로를 통해 활성화 되었다.

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Antigenotoxicity of Ginseng Petroleum Ether Extract and its Action Mechanism (인삼 지용성성분인 유전독성억제효과와 작용기전)

  • 허문영
    • Journal of Food Hygiene and Safety
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    • v.13 no.3
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    • pp.243-251
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    • 1998
  • Panax ginseng C.A. Meyer has been extensively used in the traditional oriental medicine as a restorative, tonic and prophylatic agent. Petroleum ether extract of panax ginseng C.A. Meyer (GPE) and its several fractions (PI-P5) were tested for the evaluation of antigenotoxicity against N-methyl-N-nitrosourea (MNU) and benzo(a)pyrene [B(a)P]-induced micronucleated reticulocytes in mouse peripheral blood. GPE and P2 showed more significant anticlastogenicity than other fractions did. To elucidate the anticlastogenic action mechanism of GPE and P2 against B(a)P, the alteration of B(a)P metabolism was studied. GPE and P2 inhibited B(a)P metabolism in the presence of 8-9 mix and decreased B(a)P-DNA binding in calf thymus DNA with 8-9 mix. They also decreased [$^3H$] MNU induced DNA binding and methylation to 7-methyl guanine and $O^{6}-methyl$ guanine adducts in calf thymus DNA by RPLC analysis. These results suggest that the anticlastogenicity of GPE and P2 on the B(a)P or MNU-induced clastogenicity is due to decrease of DNA binding with B(a)P or MNU, the inhibition of metabolism with B(a)P and the inhibition of methylation in DNA. Therefore, GPE and P2 may be useful chemopreventive agents of alkylating agent like MNU and secondary carcinogen like B(a)P.

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Counter anion effects in anion exchange membrane-fabricated non-aqueous vanadium redox flow battery

  • Son, Pyeong Soo;Oh, Min-Seok;Ye, Jun-Hee;Choi, Seong-Ho
    • Analytical Science and Technology
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    • v.28 no.5
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    • pp.341-346
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    • 2015
  • In order to understand the counter anionic effects in a non-aqueous vanadium redox flow battery (VRFB), we synthesized four types of electrolyte salts (1-ethyltriethamine tertafluoroborate, [E-TEDA]+[BF4], 1-ethyltriethamine hexafluorophosphate, [E-TEDA]+[PF6], 1-butyltriethylamine tertafluoroborate, [B-TEDA]+[BF4], and 1-buthyltriethamine hexafluorophosphate [B-TEDA]+[PF6]) by counter anion exchange reaction after the SN2 reaction. We confirmed the successful synthesis of the electrolyte salts [E-TEDA]+[Br] and [B-TEDA]+[Br] via 1H-NMR spectroscopy and GC-mass analysis before the counter anion exchange reaction. The electric potential of the vanadium acetylacetonate, V(acac)3, as an energy storage chemical was shown to be 2.2 V in the acetonitrile solvent with each of the [E-TEDA]+[BF4], [E-TEDA]+[PF6], [B-TEDA]+[BF4], and [B-TEDA]+[PF6] electrolyte salts. In a non-aqueous VRFB with a commercial Neosepta AFN membrane, the maximum voltages reached 1.0 V and 1.5 V under a fixed current value of 0.1 mA in acetonitrile with the [E-TEDA]+[BF4] and [E-TEDA]+[PF6] electrolyte salts, respectively. The maximum voltage was 0.8 V and 1.1 V under a fixed current value of 0.1 mA in acetonitrile with the [B-TEDA]+[BF4] and [B-TEDA]+[PF6] electrolyte salts, respectively. From these results, we concluded that in the non-aqueous VRFB more of the [PF6] counter anion than the [BF4] counter anion was transported onto the commercial Neosepta AFN anion exchange membrane.

Inhibition of Low Density Lipoprotein-oxidation, ACAT-1, and ACAT-2 by Lignans from the Bark of Machilus thunbergii

  • Shrestha, Sabina;Park, Ji-Hae;Lee, Dae-Young;Cho, Jin-Gyeong;Lee, Do-Gyeong;Cho, Moon-Hee;Jeong, Tae-Sook;Kang, Hee-Cheol;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • v.54 no.1
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    • pp.63-66
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    • 2011
  • The bark of Machilus thunbergii was extracted with 80% aqueous methanol (MeOH), and the concentrated extract was partitioned using ethyl acetate (EtOAc), butanol (n-BuOH), and $H_2O$, successively. From the EtOAc fraction, five lignans were isolated through the repeated silica gel, octadecyl silica gel (ODS) and, Sephadex LH-20 column chromatography. Based on nuclear magnetic resonance (NMR), mass spectroscopy (MS), and infrared spectroscopy (IR) spectroscopic data, the chemical structures of the compounds were determined to be machilin A (1), machilin F (2), licarin A (3), nectandrin A (4), and nectandrin B, (5). This study presents comparative account of five lignans from M. thunbergii bark contributing inhibition of low density lipoprotein (LDL), ACAT-1, and ACAT-2. Compounds 2-5 showed varied degree of antioxidant activity on LDL with $IC_{50}$ values of 2.1, 11.8, 15.3, and $4.1{\mu}M$. Compounds 1, 2, and 3 showed inhibition activity on ACAT-1 with values $63.4{\pm}6.9%$ ($IC_{50}=66.8{\mu}M$), $53.7{\pm}0.9%$ ($IC_{50}=109.2{\mu}M$), and $78.7{\pm}0.2%$ ($IC_{50}=40.6{\mu}M$), respectively, at a concentration of 50 mg/mL, and on ACAT-2 with values $47.3{\pm}1.5%$ ($IC_{50}=149.7{\mu}M$), $39.2{\pm}0.2%$ ($IC_{50}=165.2{\mu}M$), and $52.1{\pm}1.0%$ ($IC_{50}=131.0{\mu}M$, respectively, at a concentration of 50 mg/mL.

Chemical Constituents of Domestic Quercus spp. Barks (국내산 참나무속 수종 수피의 추출성분)

  • Kim, Jin-Kyu;Kwon, Dong-Joo;Lim, Soon-Sung;Bae, Young-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.38 no.4
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    • pp.359-374
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    • 2010
  • This study was carried out to investigate the chemotaxonomical correlation and chemical constituents of domestic Quercus spp. barks. The barks of Q. mongolica, Q. aliena, Q. serrata, Q. acutissima, Q. dentata, and Q. variabilis were collected in the experimental forest of Kangwon National University. The combined extracts were successively fractionated with n-hexane, methylene chloride and ethyl acetate using a separation funnel. A portion of the ethyl acetate and H2O soluble materials of each species were chromatographed on a Sephadex LH-20 column using various aqueous MeOH and EtOH-hexane as washing solvents. Spectrometric analysis such as NMR and MS, including TLC, were performed to characterize the structures of the isolated compounds. Ellagic acid (0.03 g), (+)-catechin (4.59 g), taxifolin (3.35 g), and glucodistylin (20.52 g) were isolated from Q. mongolica bark. Gallic acid (0.18 g), (+)-catechin (8.52 g), (+)-gallocatechin (0.09 g), taxifolin (0.54 g), and glucodistylin (3.28 g) were characterized from Q. acutissima bark. Gallic acid (0.38 g), ellagic acid (0.11 g), (+)-catechin (2.01 g), (+)-gallocatechin (0.12 g), and glucodistylin (0.39 g) were identified from Q. dentata bark. Ellagic acid (1.51 g), (+)-catechin (21.91 g), and glucodistylin (3.91 g) were purified from Q. aliena bark. Ellagic acid (0.84 g), (+)-catechin (0.82 g), taxifolin (4.02 g), and glucodistylin (21.50) were isolated from Q. serrata bark. Gallic acid (0.24 g), caffeic acid (0.05 g), (+)-catechin (0.32 g), and glucodistylin (0.65 g) were purified from Q. variabilis bark. (+)-Catechin and glucodistylin were isolated from all the barks. Glucodistylin can be a taxonomic index on Quercus spp.

A Tyrosinase Inhibitor Isolated from the Seeds of Eriobotrya japonica (비파 씨로부터 Tyrosinase 저해 활성물질의 분리)

  • Kim, Tae-Hoon;Shin, Seung-Ryeul;Kim, Tae-Wan;Lee, In-Chul;Park, Moon-Young;Jo, Cheo-Run
    • Food Science and Preservation
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    • v.16 no.3
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    • pp.435-441
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    • 2009
  • Activity-guided isolation from the ethylacetate (EtOAc)-soluble portion of a methanolic extract of the seeds of Eriobotrya japonica, using several bioassays, led to the isolation and identification of six phenolic compounds of previously known structure: benzaldehyde (1), chlorogenic acid (2), caffeic acid (3), benzoic acid (4), ferulic acid (5), and amygdalin (6). Of these, benzaldehyde (1) exhibited tyrosinase inhibitory activity in a bioassay. In addition, chlorogenic acid (2) and caffeic acid (3) were found to have strong antioxidative effects on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and superoxide dismutase (SOD)-like activity.