• Title/Summary/Keyword: TLC analysis

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Optimal Condition for Eicosapentaenoic Acid Production and Purification from Psychrophillic Marine Baterium Shewanella sp. L93 (호냉성 해양세균 Shewanella sp. L93로부터 Eicosapentaenoic Acid 생산 및 정제를 위한 최적화 조건)

  • Mo, Sang-Joon;Hong, Hye-Won;Bang, Ji-Heon;Cho, Ki-Woong
    • Microbiology and Biotechnology Letters
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    • v.39 no.3
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    • pp.218-223
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    • 2011
  • To obtain eicosapentaenoic acid (EPA)-producing bacteria, some 600 strains of bacteria were isolated from Antarctic sediment and marine organisms during the summer expedition of 1999-2000 and 7 EPA-producing bacteria were obtained through screening with TLC and GC. A strain designated as L93 showed the highest EPA production, which was gram-negative, rod-shaped bacterium. L93 strain was identified as Shewanella sp., from the sequence analysis of 16S rDNA. Optimal conditions temperature and pH for the growth and EPA production were about $4^{\circ}C$ and pH 7. In addition, its production was optimized by 50%(w/v) sea salt. We establish the optimal production system to produce about 320 mg per liter by using this optimal EPA production conditions. EPA-methyl ester was purified from cultured L93 strain to a purity of higher than 97% and typical purification yield is greater than 72% of the input amount via urea complexation and HPLC.

Antioxidative Materials in Domestic Meju and Doenjang 2. Separation of Lipophilic Brown Pigment and Their Antioxdative Activity (재래식 메주 및 된장중의 항산화성 물질에 관한 연구 2. 지용성 갈변물질의 분리와 항산화력)

  • Kim, Mi-Hye;Im, Sang-Sun;Kim, Sung-Hee;Kim, Gyeong-Eup;Lee, Jong-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.2
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    • pp.251-260
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    • 1994
  • Lipophilic brown pigments produced during the fermentation of domestic Meju and Doenjang were fractionated by column and thin -layer chromatography (TLC). Each of the fractions was tested for the antioxidant activity and then characterized by spectroscopic analysis. The lipophilic brown pigments were separated into chloroform -soluble and methanol-soluble parts in which Meju resulted the higher content of chloroform-soluble part than that of methanol-soluble part ; however, Doenjang exhibited the opposite result to that of Meju. More strong antioxidant activity was found in the methanol-soluble part than the chloroform-soluble part. Four and five fractions were separated from chloroform-soluble and methanol-soluble parts respectively. by silicagel TLC. The fraction that exhibited the high antioxidant activity showed a strong absorption at 260nm caused by amino compounds in UV spectrum The other fractions which did not have antioxidant activity absorbed at 240nm by carbonic acid and it ester,. IR spectrum of each fraction commonly showed absorption at 3400cm-1 , 2800cm-1 , 1700cm-1, 1600cm-1, 1400cm-1 , 1300cm-1 and 1100cm-1. Especially , the fraction which had a strong antioxidant activity showed absorption at 2800cm-1, 1400cm-1, 1600cm-1 suggesting that the fraction contain Schiff's base and primary amine structure.

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Modulation of the Regioselectivity of a Thermotoga neapolitana $\beta$-Glucosidase by Site-Directed Mutagenesis

  • Choi, Ki-Won;Park, Kyung-Min;Jun, So-Young;Park, Cheon-Seok;Park, Kwan-Hwa;Cha, Jae-Ho
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.901-907
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    • 2008
  • Thermotoga neapolitana $\beta$-glucosidase (BglA) was subjected to site-directed mutagenesis in an effort to increase its ability to synthesize arbutin derivatives by transglycosylation. The transglycosylation reaction of the wild-type enzyme displays major ${\beta}(1,6)$ and minor ${\beta}(1,3)$ or ${\beta}(1,4)$ regioselectivity. The three mutants, N291T, F412S, and N291T/F412S, increased the ratio of transglycosylation/hydrolysis compared with the wild-type enzyme when pNPG and arbutin were used as a substrate and an acceptor, respectively. N291T and N219T/F412S had transglycosylation/hydrolysis ratios about 3- and 8-fold higher, respectively, than that of the wild-type enzyme. This is due to the decreased hydrolytic activity of the mutant rather than increased transglycosylation activity. Interestingly, N291T showed altered regioselectivity, as well as increased transglycosylation products. TLC analysis of the transglycosylation products indicated that N291T retained its ${\beta}(1,3)$ regioselectivity, but lost its ${\beta}(1,4)$ and ${\beta}(1,6)$ regioselectivity. The altered regioselectivity of N291T using two other acceptors, esculin and salicin, was also confirmed by TLC. The major transglycosylation products of the wild type and N291T mutant were clearly different. This result suggests that Asn-291 is highly involved in the catalytic mechanism by controlling the transglycosylation reaction.

Isolation and Characterization of Bacillus sp. P16 Producing Extracellular Chitosanase (키토산분해효소를 생산 분비하는 Bacillus sp. P16의 선발 및 특성)

  • Jung, Mi-Ra;Jo, Yoo-Young;Chi, Yeon-Tae;Park, Ro-Dong
    • Applied Biological Chemistry
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    • v.40 no.5
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    • pp.369-374
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    • 1997
  • An endochitosanase-producing bacterium was isolated from soil and identified as a strain of Bacillus sp. The isolate was gram positive, rod shape $(0.4-0.6{\times}1.6-2.2{\mu}m)$, endospore-forming, catalase positive, and mobility positive, and grown at pH 4.5-11.0 and upto $42^{\circ}C$ in the medium containing 2% NaCl. RAPD analysis of the DNA purified from the strain was also performed, and the chitosanase-producing strain was named as Bacillus sp. P16. The culture supernatant of the strain showed strong liquefaction activity and rapidly decreased viscosity of chitosan solution. By TLC and HPLC, chitooligosaccharides of DP 2-7 were separated and identified from the enzyme hydrolyzates of chitosan. The chitosanase from Bacillus sp. P16 was thus regarded as an endo-splitting type.

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Screening of Antagonistic Actinomycetes for Potato Scab Control and Isolation of Antibiotic Compound (감자 더뎅이병원균에 대해 길항활성을 갖는 방선균 탐색 및 항균 활성물질의 분리)

  • Lee, Hyang-Burm;Cho, Jong-Wun;Lim, Chi-Hwan;Kim, Chang-Jin
    • Applied Biological Chemistry
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    • v.47 no.2
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    • pp.164-169
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    • 2004
  • In the course of our screening for biocontrol agent (BCA) against Streptomyces scabiei and S. turgidiscabies causing potato scab using 5,000 actinomtcete isolates, 9 antagonistic strains were selected as BCA candidates through in vitro and in vivo assay. An antagonistic strain, A020645 was highly resistant to some pesticides and antibiotics such as dazomet and mancozeb and showed high control value in vivo. Two bioactive compounds (compound A, B) were purified by anion exchange chromatography, solid phase (ODS) extraction, TLC and reverse phase HPLC. Their chemical structures are now thought to be nucleoside derivative as determined by $^1H-NMR$ data analysis. Their full chemical structures would be elucidated through $^{13}C-NMR$, HMQC and HMBC analyses. Further studies will be focused on fitness in soil and formulation of the BCA candidates.

Isolation of Herbicidal Substances from Bulbs of Lycoris flavescens M.Y.Kim & S.T.Lee (붉노랑상사화 인경으로부터 살초활성 물질의 분리)

  • Lee, Dong-Gu;Kim, Kun-Woo
    • Korean Journal of Weed Science
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    • v.31 no.4
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    • pp.330-339
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    • 2011
  • This study was conducted to determine the herbicidal activity of herbicidal substances and identify them in bulbs of Lycoris flavescens. Methanol extract was purified by a series of chromatographic techniques including silica gel column chromatography, preparative TLC, and HPLC. The final HPLC gave two active fractions and two herbicidal substances were obtained. By GC/MS analysis, one was identified as galanthine (galanthan-1-ol) and the other was identified as montanine ($O^2$-methyl pancracine), an isoquinoline alkaloid. Montanine showed nearly 100% of growth inhibition on the shoot and root of barnyardgrass (Echinochloa crusgalli) seedlings at $20{\mu}g\;mL^{-1}$ as compared with the control. Meanwhile, methanol extract of L. flavescens bulbs showed only about 3.1% and 8.3% of growth inhibition on the shoot and root of rice cultivar, Hwayeongbyeo seedlings at $1,000{\mu}g\;mL^{-1}$ as compared with the control, respectively.

Study on antitumor activity of Chinese and Korean Oldenlandiae Herba and its effective compound (중국 및 국산 백화사설초의 항암활성과 지표물질 연구)

  • Lee Hyo Jeong;Song Gyu Young;Ra Jeong Chan;Ahn Kyoo Seok;Kim Sung Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.4
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    • pp.1059-1064
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    • 2003
  • Oldenlandiae Herba has been used for the prevention or treatment of cancer in oriental medicine for years. Experimentally its antitumor activity was reported. However, in order to develop new Korean original plants. we tried to study the antitumor activity of Chinese and Korean origin Oldenlandiae Herba. We first evaluated the cytotoxicity of Oldenlandiae Herba originated from China and Korea on HT1080, U937 and SK-mel tumor cells. MeOH extract of Chinese Oldelandiae Herba was more effective than MeOH extract of Korean Oldenlandiae Herba. Approixmately IC50 of two species of Oldelandiae Herba was 380-450 ug/ml. For evaluation of antiangiogenic activity, proliferation assay with HUVECs(Human umblical vein endothelial cells) was done with three samples. Chinese Herba and Korean Herba and root. Korean root and herba was more effective than Chinese Herba. Thus, we found Korean Oldenlandiae was more effective on angigenic activity than Chiness Oldenlandiae. In an anlytical study on effective compounds from Oldenlandiae, the spot of ursolic acid was more marked by Korean Oldenlandiae than by Chinese Oldenlandiae. whereas the spot of asperuloside was more pronounced by Chinese Oldenlandiaethan by Korean Oldenlandiae by TLC analysis.

Extractives from the needles of Juniperus rigida Siebold et Zucearin and Antioxidant activity (노간주나무(Juniperus rigida Siebold et Zucearini) 잎의 추출성분 및 항산화 활성)

  • Lee, Sang-Keug;Kim, Jin-Kyu;Ham, Yeon-Ho;Bae, Young-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.32 no.1
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    • pp.59-66
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    • 2004
  • The dried neeldes(1.5 kg) of Juniperus rigida were ground, extracted with acetone-H2O(7:3, v/v), concentrated, and fractionated with a series of hexane, CH2Cl2, EtOAc and water on a separatory funnel. Each fraction was freeze dried to give dark-brown powder and EtOAc and water soluble fractions were chromatographed on a Sephadex LH-20 column using a series of aqueous methanol and ethanol-hexane mixture as eluent. Spectrometric analysis such as NMR and FAB or EI-MS including TLC were performed to characterize the structures of the isolated compounds. The needles of Juniperus rigida contained a large amount of (+)-catechin, quercetin-3-O-α-L-rhamnopyranoside and isoconiferin, in addition to a small amount of umbelliferone and quercetin-3-O-β-D-rutinoside. The antioxidative activities of each fraction and isolated compounds were tested by DPPH radical scavenging method, and EtOAc soluble fraction, (+)-catechin and quercetin-3-O-α-L-rhamnopyranoside were effective.

Evaluation of Immunomodulatory and Biological Effects of Aquilaria crassna Extracts (침향 추출물의 면역조절 및 생리활성 분석)

  • You-Lim, Hwang;Kwang-Youn, Kim;Sun Nyoung, Yu;Kwang-Il, Park;Soon-Cheol, Ahn
    • Herbal Formula Science
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    • v.30 no.4
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    • pp.249-257
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    • 2022
  • Objectives : Aquilaria crassna is a traditional herbal medicine, which is used to treat allergies, diabetes, neurological diseases. Recently, Aquilaria crassna extracts have been reported in anti-bacterial and anti-inflammatory activities. In this study, various solvents fraction of Aquilaria crassna were investigated on various physiological activities. Methods : According to the polarity, the solvents fraction of Aquilaria crassna were confirmed through TLC, and the activities of the extracts were confirmed in anti-diabetes, anti-obesity, whitening, anti-gout, and anti-inflammation. Results : TLC results showed that ACM and ACM/E have similar patterns and most of the components were transferred to ACM/E. Treatment with ACM and ACM/E fraction were significantly decreased the generation of NO in lipopolysaccharide (LPS)-stimulated macrophage cells. Analysis of biological activities such as α-glucosidase, protein tyrosine phosphatase (PTP1B), tyrosinase, xanthine oxidase (XO) and pancreatic lipase inhibition, showed that ACM and ACM/E have more inhibitory effects than other fractions. Conclusions : Therefore, the results of the present study clearly demonstrate that Aquilaria crassna and its constituents might be beneficial in the prevention or treatment of immune-regulating effects.

Antioxidative Activity and Component Analysis of Psidium guajava Leaf Extracts (구아바 잎 추출물의 항산화 활성과 성분 분석)

  • Yang, Hee-Jung;Kim, Eun-Hee;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.3
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    • pp.233-244
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    • 2008
  • In this study, the antioxidative effects, inhibitory effects on elastase and tyrosinase, and component analysis of Psidium guajava leaf extracts were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities $(FSC_{50})$ of extract/fractions of Psidium guajava leaf were in the order: 50% ethanol extract $(7.05{\mu}g/mL)$ < ethyl acetate fraction $(3.36{\mu}g/mL)$ < deglycosylated flavonoid aglycone fraction $(3.24{\mu}g/mL)$. Reactive oxygen species (ROS) scavenging activities $(OSC_{50})$ of some Psidium guajava leaf extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activities were 50% ethanol extract $(OSC_{50},\;2.17{\mu}g/mL)$ < ethyl acetate fraction $(0.64{\mu}g/mL)$ < deglycosylated flavonoid aglycone fraction $(3.39{\mu}g/mL)$. Aglycone fraction showed the most prominent ROS scavenging activity. The protective effects of extract/fractions of Psidium guajava leaf on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Psidium guajava leaf extracts suppressed photohemolysis in a concentration dependent manner $(1{\sim}10{\mu}g/mL)$, particularly deglycosylated flavonoid aglycone fraction exhibited the most prominent celluar protective effect ${\tau}_{50}\;107.5min\;at\;1{\mu}g/mL)$. Aglycone fraction obtained from the deglycosylation reaction of ethyl acetate fraction among the Psidium guajava leaf extracts, showed 1 band in TLC and 1 peak in HPLC experiments (360 nm). One component was identified as quercetin. TLC chromatogram of ethyl acetate fraction of Psidium guajava leaf extract revealed 5 bands and HPLC chromatogram showed 5 peaks, which were identified as quercetin 3-O-gentobioside (10.32%) , quercetin 3-O-${\beta}$-D-glucoside (isoquercitin, 13.30%), quercetin 3-O-${\beta}$-D-galactoside (hyperin, 11.34%), quercetin 3-O-${\alpha}$-L-arabinoside (guajavarin, 19.70%), quercetin 3-O-${\beta}$-L-rhamnoside (quercitrin, 45.33%) in the order of elution time. The inhibitory effect of Psidium guajava leaf extracts on tyrosinase were investigated to assess their whitening efficacy. Finally, their anti-elastase activities were measured to predict the anti-wrinkle efficacy in the human skin. Inhibitory effects $(IC_{50})$ on tyrosinase of some Psidium guajava leaf extracts was 50% ethanol extract $(149.67{\mu}g/mL)$ < ethylacetate fraction $(30.67{\mu}g/mL)$ < deglycosylated aglycone fraction $(17.10{\mu}g/mL)$. Inhibitory effects $(IC_{50})$ on elastase of some Psidium guajava leaf extracts was 50% ethanol extract $(6.60{\mu}g/mL)$ < deglycosylated aglycone fraction $(5.66{\mu}g/mL)$ < ethylacetate fraction $(3.44{\mu}g/mL)$. These results indicate that extract/fractions of Psidium guajava leaf can function as antioxidants in bioloigcal systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. And component analysis of Psidium guajava leaf extract and inhibitory activity on elastase of the aglycone fraction could be applicable to new functional cosmetics for smoothing wrinkles.