• Title/Summary/Keyword: ninhydrin

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Purification and Characteristics of Tyrosinase Inhibitor Produced by Actinomycetes F-97 (방선균 F-97이 생산하는 Tyrosinase 저해제의 정제 및 특징)

  • Bang, Byung-Ho;Rhee, Moon-Soo;Kim, Jin-O;Yi, Dong-Heui
    • Applied Biological Chemistry
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    • v.51 no.3
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    • pp.153-158
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    • 2008
  • An actinomycetes F-97 producing tyrosinase inhibitor was isolated from soil samples. Isolation and purification of tyrosinase inhibitor produced by F-97 was performed as follows: IRC-120 ($NH_4^+$ type) column chromatography, silica gel column chromatography, $C_{18}$ column chromatography and Sephadex LH-20 column chromatography were used successively after the centrifuged supernatant was adjusted to pH 4.0. To identify the purity of the inhibitor, octadecylsilyl(ODS) HPLC was carried out with 5% methanol as a mobile phase. Finally, the purification yield of a tyrosinase inhibitor was 5.24%. The inhibitor was very soluble in water, methanol and ethanol but insoluble in acetone, butanol, ethylacetate and chloroform. The ${\lambda}_{max}$ value of this inhibitor in water was 194nm under UV light. The biochemical test of the inhibitor was positive in Molish, Benedict, cone. $H_2SO_4$, and $KMnO_4$ tests but negative in iodine, ninhydrin, Million, Sakaguchi, xanthoproteic and Emerson tests. The tyrosinase inhibitor was stable against heat treatment of $100^{\circ}C$ for 50 minutes and pH $4{\sim}9$. The $IC_{50}$ value of this inhibitor was $19.2{\mu}g/ml$ for mushroom tyrosinase. In $1,000{\mu}g/ml$ inhibitor concentration, inhibition zone was 27 mm for Streptomyces bikiniensis NRRL B-1049. The inhibition of F-97 against mushroom tyrosinase was competitive with tyrosine.

Free Amino Acids of Xylem-Pith in Panax ginseng Root. (인삼근동체중심부의 유리아미노산)

  • Lee, Mee-Kyoung;Park, Hoon
    • Journal of Ginseng Research
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    • v.11 no.1
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    • pp.32-38
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    • 1987
  • Composition of free amino acids (FAA) in the central part (xylem plus piths,of tap root in P. ginseng was investigated in velation to stem status at harvest. The sum of FAA tended to be higher with dead stem than with healthy one but both were not significantly different. The sum of FAA (3.6-4.9% dried weight) was much less than total FAA, suggesting that water soluble nonprotein fraction contained large quantity of ninhydrih positive components except FAA. Pattern of amino acid composition between both stem status was not different. Ten of all 17 amino acids showed increasing tendency with dead stem and two, glutamic acid and cysteine, decreasing. Major FAA were arginine (relative content 58%), glycine (8.2), lysine (5.9), serine (5.7), glutamic acid (4.2) and aspartic acid (3.5). Above facts strongly suggest that the inside white of red ginseng did not closely related with FAA and that early defoliation or stem death did not decrease FAA. The content of arginine was heighest in all cases reported indicating the important role of nitrogen metabolism. Pattern of PAA composition except arginine was not different in present samples but greatly different with other cases reported mainly due to alanine, phenylalanine, glycine and proline.

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Antioxidant Activity of Panax Ginseng Browning Products (고려인삼(高麗人蓼) 갈변물질(褐變物質)의 항산화효과(抗酸化效果))

  • Kim, Sang-Dal;Do, Jae-Ho;Oh, Hoon-Il
    • Applied Biological Chemistry
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    • v.24 no.3
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    • pp.161-166
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    • 1981
  • Products of ginseng browning reaction were investigated to determine the nature of their antioxidant activity using a model system, White Ginseng and Red Ginseng extracts. In the simulated ginseng model system, the brown color was intensified with an increase in the length of reaction time and the antioxidant activity initially increased in proportion the length of reaction time for up to 17 hrs and then leveled off thereafter. Pararell results were obtained manufacturing of Red Ginseng, Comparison of the antioxidant activity of the inner and outer solutions after dialysis of browning solution showed that the outer solution had a stronger antioxidant activity than the inner one. For further analyses, browning reaction products were fractionated into three peaks on Amberlite CG-120 type I ionexchange resin and designated as fractions I, II and III in order of elution. Partial characterization of the fractions revealed that floe most intense brown fraction (Fraction II) had the strongest antioxidant activity and also exhibited reducing power for Somogyi-Nelson reagents and ninhydrin positive reaction. Both the browning reaction products and Red Ginseng extracts were found to possess potent reactivity with ${\alpha}$,${\alpha}'$-diphenyl-${\beta}$-picrylhydrazyl, whereas White Ginseng extracts showed negligible reactivity.

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Toxicity and Characteristics of Antifungal Substances Produced by Bacillus amyloliquefaciens IUB158-03 (Bacillus amyloliquefaciens IUB158-03이 생산하는 항진균물질의 생화학적 특성 및 독성)

  • Kim, Hye-Young;Lee, Tae-Soo
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1672-1678
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    • 2009
  • The purified antifungal substances produced by Bacillus amyloliquefaciens IUB158-03 was positive to ninhydrin but negative to aniline, suggesting that the antifungal substance could be a peptide. FAB-MS, UV adsorption spectrum, and amino acid composition analysis revealed that the molecular weight of the antifungal substance was 1042 and that maximal adsorption was at 220 nm and 277 nm. The antifungal substance was composed of $Asn_3$, $Gln_2$, $Ser_1$, $Gly_1$, and $Tyr_1$. The composition and structural characteristics of antifungal substance were analysed by $^1H$-NMR spectrum, $^1H$-COSY, HMQC, which revealed that the compound belongs to the iturin A family. Temperature and pH had little effect on the stability of the antifungal substance in the ranges of $-70{\sim}121^{\circ}C$ and pH 6.0~10.0, respectively. It showed strong antibiotic activity against fungi. An in vitro cytotoxicity test using NIH3T3 cell showed that the antifungal substance does not have cytotoxicity. The number of circulating leukocytes and the hematobiological analysis of the mice administered with the antifungal substances was similar to those of the control group, indicating no cytotoxicity in vivo. Therefore, the antifungal substances extracted from culture broth of Bacillus amyloliquefaciens IUB158-03 have future potential as biocontrol agents against plant diseases caused by fungi.

Antioxidative Effect and Characteristics of Different Model Melanoidins with Same Color Intensity (색도를 동일하게 조정한 Model Melanoidin들의 항산화효과 및 특성)

  • Lim, Won-Yong;Kim, Jong-Sang;Moon, Gap-Soon
    • Korean Journal of Food Science and Technology
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    • v.29 no.5
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    • pp.1045-1051
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    • 1997
  • Three kinds of model melanoidins adjusted to have the same brown color intensity were made from glucose-glycine, glucose-lysine, xylose-arginine and their antioxidative properties were determined. The antioxidative activities of these model melanoidins in linoleic acid emulsion system were determined by ferric thiocyanate method, conjugated diene contents, peroxide value and electron donating ability by DPPH. Xylose-arginine melanoidin showed the strongest antioxidative activity and electron donating ability. The antioxidative effect of melanoidin could be reliably predicted by determining peroxide value and DPPH method. Each melanoidin was separated on Sephadex G-50 column, and brown color intensity, reducing power, ninhydrin positive reaction and antioxidative activity of each fraction were determined. The antioxidative activities of melanoidin fractions showed strong correlation with their brown color intensity and especially to their reducing power. In spite of same brown color intensity, there is no big differences between these model melanoidins, thus xylose-arginine showing strongest antioxidative activity followed by glucose-lysine and glucose-glycine melanoidin. Xylose-arginine melanoidin also showed the strongest electron donating activity and broad range of reducing power when fractionated on Sephadex G-50, which was different tendency from the other model melanoidin.

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Constituents of Antimutagenic Factor from Brown Rice (현미의 항돌연변이 활성물질의 구성성분)

  • Chun, Hyang-Sook;Moon, Tae-Wha;Kim, In-Ho
    • Applied Biological Chemistry
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    • v.38 no.5
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    • pp.478-483
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    • 1995
  • To investigate the constituents of antimutagenic factor from brown rice, methanol extracts were fractionated into ether, ethylacetate, buthanol and aqueous fractions. The ether fractions showed distinct antimutagenic effect and active spot were selected by silica gel chromatography. The specific activity of active spot decreased with isolation of the active components from the methanol extract. Qualitative analyses of the active spot by using various spraying reagents revealed that ninhydrin and orcinol did not develop colored reactions. But, ferric chloride, 2,7-dichlorofluorescein, antimony pentachloride, phosphomolybdic acid, bromothymol blue and rhodamine 6G led to colored reactions. These results suggested that the consitituents of active material were neither polar nor nitrogen-containing compounds and that they may contain phenolic compounds and fatty acid derivatives. Main compounds of the active spot were analyzed to be o-hydroxy benzyl alcohol(saligenin), octanoic acid(caprylic acid), 9,12-cis-octadecadienoic acid(linoleic acid), 11-cis-octadecenoic acid(oleic acid), hexadecanoic acid(palmitic acid), 1H-indole-2-carboxylic acid and 1,2-benzenedicarboxylic acid(phthalate) in GC/Mass spectrum, and antimutagenicity of these active compounds using standard regeant was reconfirmed in S. typhimurium reversion assay.

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Drug Delivery Study on Chitosan Nanoparticles Using Iron Oxide (II, III) and Valine (Iron Oxide(II, III)와 Valine을 이용한 키토산 나노입자의 약물전달 연구)

  • Jang, So-Hyeon;Kang, Ik-Joong
    • Korean Chemical Engineering Research
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    • v.59 no.4
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    • pp.514-520
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    • 2021
  • A drug delivery system (DDS) based on nanoparticles has been used as a mediator to improve the efficacy of a drug by controlling the amount of drug released and delivering it to a target place. Chitosan, which is non-toxic and biodegradable, has good biocompatibility and excellent adsorption, so it can be used as a drug delivery vehicle. Valine, the essential amino acids, helps muscle growth and tissue recovery, and along with other amino acids. It lowers blood sugar levels and increases growth hormone production. In this study, Valine was adsorbed on magnetic chitosan which is capable of drug absorption, and Fe3O4-Valine CNPs was prepared through cross-linking with TPP (Tripolyphosphate). And then absorption and release trends of valine were investigated with the Fe3O4-Valine CNPs. Fe3O4, which has relatively high stability, is used to make the drug carrier magnetic so that the drug can be delivered to a target place. At optimal conditions, the absorption and release tendency of Fe3O4-Valine CNP was confirmed by analyzing by UV-Vis through the Ninhydrin test which is the color reaction of amino acids and by measuring the size of the particles, it was confirmed that it is suitable as a drug carrier.

Ultrastructure of the digestive tract of Korean Leech (Erpobdella lineata) (한국산 거머리 (Erpobdella lineate) 소화관의 미세구조)

  • Chang, N.S.
    • Applied Microscopy
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    • v.24 no.3
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    • pp.34-45
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    • 1994
  • This investigation has been carried out to examine the structure of digestive tract from Korean Leech, Erpobdella lineata, using light and electron microscope. The digestive tract is composed of mouth, pharynx, Oesophagus, six-chambered stomach, three-chambered intestine, rectum and anus. Stomach and intestine have not gastric or intestine ceca and consist of only straight tube. All digestive tracts from pharynx to rectum are covered with simple columnar epithelial cells. While the surfaces of endothelial cell of pharynx and rectum are covered with cuticular layer of about $0.3{\mu}m$ in thickness, stomach and intestine are covered with estimated $0.2-0.3{\mu}m$ and $0.5{\mu}m$ microvilli respectively. Circular folds were found only in first and second chambers of stomach, intestine and rectum, but not in pharynx and the other chambers (third to sixth) of stomach. The granules of $0.3-0.8{\mu}m$ and $0.5-1.0{\mu}m$ in diameter were observed in the cytoplasm of stomach endothelial cell. These granules were demonstrated to contain protein which showed a positive reaction to ninhydrin. It was also found that there are well-developed microvilli in the apical portion of intestine endothelial cell in which endocytosis occurs actively.

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Antibiotics produced by anaerobic fermentation of Streptococcus sp. An-21-1 isolated from domestic soil, Fermentation and purification of antibiotics from anaerobe (국내토양에서 분리한 혐기성 세균 Streptococcus sp. An-21-1 이 생성하는 항생물질 II. 항생물질을 생성하는 혐기성 세균의 발효 및 항생물질의 분리 정제)

  • Park, Seung-chun;Yun, Hyo-in;Oh, Tae-kwang
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.61-69
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    • 1993
  • In order to search for new antibiotics from anaerobic bacteria, a large number of samples from domestic soil were collected and processed by apropriate methods. A potential strain, Streptococcus sp. An-21-1, was found to produce antimicrobial compounds. The Results were as follows; 1. During fermentation, the bacteria grew rapidly up to 20hr, thereafter entered the death phase. The optimal temperature and pH for the bacterial growth were $37^{\circ}C$ and pH 7.0, respectively. 2. Antibiotics were purified from culture broth by solvent extraction, silica gel column chromatography and Sepadex L.H 20 column. 3. Physicochemical properties of Ap-1 and Ap-2 were similar ; Their melting points were between $234-237^{\circ}C$. Color reactions of ninhydrin, 2,7-dichlorofluorescein, 4-dimethylaminobenzaldehyde, Dragendroffs reagent and 20% $H_2SO_4$, were positive. Therefore, we assumed that these antibiotics have amine group, immine group, alkaloid, and lipid components. These were stable to heat. UV spectrophotometry showed two peaks at 210 nm and 260 nm. From above results, we assumed these antibiotics are belong to the peptide antibiotic family.

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A physiological study on Sporulation of Rhizopus nigricans (Rhizopus nigricans의 포자형성에 관한 생물학적 연구)

  • 윤경하;이영록
    • Korean Journal of Microbiology
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    • v.17 no.2
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    • pp.81-93
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    • 1979
  • The mycelium of Rhizopus nigricans was harvested at intervals during the sporulation periods, fractioned into various cell components and analyzed the con!eiits of various cell materials in order to clarify the optimum conditions of sporulation and some characteristics of the metabolism during tke sporulation periods. The changes in enzyme activities, such as amylase and protease, were also measured during the sporulation period,. 1. Mycelium in distilled water culture, as control, did not sporulate but mycelial mat cultured in Petridish without mutrient spourulated. Optimum temperature range for sporulation was $20{\sim}25^{\circ}C$. 2. During the sporulation and maturation periods, proteins, especially alkali-labile protein were decreased remarkably but free amino acid and ninhydrin reactive substances in acid soluble fraction were increased, compared with control. 3. Acid solable polyphosphate was decreased but acid insoluble polyphosphate was increased, during the sporulation. 4. Carbohydrate and hexosamine in acid soluble fraction were increased, while carbohydrate in alkali insoluble residual fraction was decreased during the sporulation periods. 5. Amounts of UV-absorbing material in deoxyribonucleic acid fraction was increased a little but those in ribonucleic acid fraction was decreased, compared with control. 6. Intracellular amylases and proteases activities insporulating mycelial mat were increased continuously during the sporulation and maturation periods.

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