• Title/Summary/Keyword: reversed phase

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The Antimicrobial Characteristics of McSSP-31 Purified from the Hemocyte of the Hard-shelled Mussel, Mytilus coruscus (참담치(Mytilus coruscus) 혈구(hemocyte)에서 분리한 McSSP-31의 항균 특성 분석)

  • Oh, Ryunkyoung;Lee, Min Jeong;Kim, Young-Ok;Nam, Bo-Hye;Kong, Hee Jeong;Kim, Joo-Won;Park, Jung-Youn;Seo, Jung-Kil;Kim, Dong-Gyun
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1276-1289
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    • 2017
  • This study isolated and purified the antimicrobial peptide McSSP-31 from an acidified hemocyte extract of a Mytilus coruscus. The antimicrobial peptide was purified by using a $C_{18}$ reversed-phase high-performance liquid chromatography (HPLC). The peptide was determined to be 3330.549 Da by matrix assisted-laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS). The N-terminus of a 14 amino-acid sequence was identified as P-S-P-T-R-R-S-T-S-R-S-K-S-R by Edman degradation method. The acquired sequence showed a 93% similarity with the sperm-specific protein Phi-1, which is from M. californianus. The identified open-reading frame (ORF) of peptide was 306 bp encoding 101 amino acids, which was analyzed by rapid amplification of cDNA ends (RACE), cloning and sequencing analysis. We compared the full sequence with other known proteins that reveal the sperm-specific protein Phi-1 (93.5%) of M. californianus. Synthesized antimicrobial peptide (McSSP-31) showed antibacterial activity against gram-positive bacteria including B. subtilis, S. mutans, S. aureus and gram-negative bacteria including E. coli, K. pneumoniae, P. mirabilis, P. aeruginosa and fungi, C. albicans. Also, synthesized peptide showed strong antibacterial activity against antibiotic-resistant strains, including S. aureus. The cytotoxicity of the peptide was determined by using the HUVEC human cell line. The peptide did not exhibit any significant cytotoxic effects on the normal human cell line, and it had very low hemolytic activity with flounder hemoglobin. The results demonstrated that peptide purified from the hemocyte of a M. coruscus exhibits antibacterial activity against various bacteria and has the potential to be an alternative antibiotic agent.

A Study on Contents of Vitamin K1 in Local Agricultural Products (지역농산물의 비타민 K1 함량 조사)

  • Lee, A Reum;Kim, Joo Hee;Park, Jae-Ho;Kim, Youngho;Hong, Eui Yon;Kim, Haeng-Ran;Choi, Youngmin;Lee, Junsoo;Eom, Hyun-Ju
    • The Korean Journal of Food And Nutrition
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    • v.29 no.3
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    • pp.301-306
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    • 2016
  • Vitamin K (phylloquinone) is an essential cofactor in the synthesis of active blood-clotting factors II, VII, IX and X. Deficiency of vitamin K leads to inadequate activity of these factors, resulting in bleeding. In this study, we investigated vitamin $K_1$ content of agricultural products that are widely and specifically grown in Korea including 9 leaves and vegetables, 16 fruits, and 11 cereals and specialty crops. Vitamin $K_1$ analysis of the agro-samples was by a validated, modified, reversed phase-HPLC method with fluorescence detection after post-column derivatization. The vitamin $K_1$ content ranged from 1.83 to $682.73{\mu}g$/100 g in leaves and vegetables, 0.17 to $28.22{\mu}g$/100 g in fruits, and ND to $279{\mu}g$/100 g in cereals and specialty crops. Among the 36 samples, high content of vitamin $K_1$ were found in Gugija (Lycium chinense Miller) leaves (average $682.73{\mu}g$/100 g) and Hansan ramie leaves (average $423.12{\mu}g$/100 g); however, mushroom, amaranth and Chinese artichoke showed no detectable levels. The results of ourstudy provide reliable vitamin $K_1$ content of Korean grown agricultural products that expand nutritional information and food composition database.

Enzymatic Interesterification and Melting Characteristic for Asymmetric 1,2-Distearoyl-3-Oleoyl-rac-Glycerol Triacylglycerol Enriched Product (효소적 반응을 이용한 비대칭형 1,2-Distearoyl-3-Oleoyl-rac-Glycerol 혼합물의 생성 및 융점 특성)

  • Kim, Jin Young;Lee, Ki Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.1
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    • pp.93-101
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    • 2014
  • Asymmetric 1,2-distearoyl-3-oleoyl-rac-glycerol (SSO) triacylglycerol (TAG) is used as a cocoa butter replacer (CBR). In this study, it was produced by lipase-catalyzed interesterification of fully hydrogenated soybean oil (FHSBO) and oleic ethyl ester (OEE) in a batch type reactor at $75^{\circ}C$, 250 rpm. Different molar ratios (FHSBO : OEE=1:1, 1:2 and 1:3, w/w) and various reaction times (1, 2, 3, 4, and 5 hr) were also tested. The optimized condition for SSO was a FHSBO : OEE molar ratio of =1:1 at reaction times of 2, 3, 4, and 5 hr. Enzymatic synthesis generated SSO/SOS, as well as the other TAGs (e.g., PSO/POS, SOO/OSO, SSS), ethyl esters, monoacylglycerol (MAG), and diacylglycerol (DAG). After scale-up, fractionation by solvent (methanol and acetone) fractionation and column chromatography was applied. To reduce ethyl esters, high-melting TAGs (e.g., SSS), and SOO/OSO in reactants, solvent fractionation was applied. Using a silica gel column (sample : silica gel=2:1, wt%), MAG and DAG were removed at $25^{\circ}C$. The major fatty acid composition of the final products (with a high SSO/SOS content) was palmitic acid (C16:0, 10.9~12.9 area%), stearic acid (C18:0, 52.2~54.9 area%), and oleic acid (C18:1, 34.2~35.5 area%). In reversed-phase HPLC analysis, the major TAG species of the final product (FHSBO : OEE=1:1, 2 hr) were SSO/SOS (82.31 area%) and PSO/POS (14.51 area%). Based on the $[SS]^+$ : $[SO]^+$ ratio obtained by RP-HPLC/APCI-MS, the final product had a higher SSO (AAB type TAG) content than cocoa butter (CB). The solid fat index (SFI) of CB and the final product obtained were similar with a narrow melting point range around ~32 to $35^{\circ}C$.

Comparative Analysis of Catechins and Antioxidant Capacity in Various Grades of Organic Green Teas Grown in Boseong, Korea (보성산 유기농 녹차의 품질에 따른 카테킨 함량과 항산화능 비교 분석)

  • Park, Kyung-Ryun;Lee, Sang-Gil;Nam, Tae-Gyu;Kim, Young-Jun;Kim, Young-Rok;Kim, Dae-Ok
    • Korean Journal of Food Science and Technology
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    • v.41 no.1
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    • pp.82-86
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    • 2009
  • The objective of this study was to evaluate the effect of various solvents on extraction of bioactive phenolics and to analyze the antioxidant capacity and contents of individual catechins in various grades of green teas organically grown in Boseong, Korea. The organic green teas, based on their harvest seasons, were categorized into five grades such as Woo-Jeon, Se-Jak, Jung-Jak, Dae-Jak, and coarse tea. Solvents used to extract phenolics from these teas included water at $23^{\circ}C$ and $70^{\circ}C$ as well as 80% (v/v) aqueous methanol and ethanol. In general, aqueous organic solvents of methanol and ethanol led to higher extraction yields of phenolics than water at $23^{\circ}C$ and $70^{\circ}C$. Total phenolics and antioxidant capacity of the teas extracted with the aqueous organic solvents were approximately 1.5 to 3.2 and 1.8 to 3.8 times higher than those with water at $23^{\circ}C$ and $70^{\circ}C$, respectively. Coarse tea, the lowest grade of green tea, showed approximately 30-60% lower total phenolics and antioxidant capacity compared with the higher grade ones. Reversed-phase HPLC analysis was performed quantitatively to identify individual catechins, gallic acid, and caffeine in teas extracted with 80% (v/v) aqueous methanol. Based on their dry weights, the organic green teas contained about 1.7 to 2.9% of caffeine. Content (mg/g dry weight) of tea catechins decreased in the following order: Woo-Jeon (155.4) > Se-Jak (147.7) > Jung-Jak (143.2) > coarse tea (135.1) > Dae-Jak (130.5). (-)-Epigallocatechin gallate was the most abundant among the catechins analyzed. The highest grade of green tea, Woo-Jeon, had the highest amount of (-)-epigallocatechin gallate at 77.4 mg/g dry weight. Overall, the higher grade of organic green teas tended to have the higher level of antioxidant capacity and catechins.

Isolation and Characterization of Elastase Inhibitor from Areca catechu (빈랑으로부터 Elastase 저해물질의 분리 및 특성조사)

  • 조중제;이건국;조병기;최정도
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.26 no.1
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    • pp.163-186
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    • 2000
  • We have previously screened 150 medicinal plants on the inhibition of elastase and found a significant inhibitory effects of the extracts of Areca catechu L. on the aging and inflammation against the skin tissues. To isolate and identify the compounds having biological activity, we was further purified by each of the solvent fractions, silica gel column chromatography, preparative TLC and reversed-Phase HPLC. Peak in HPLC, which coincided with the inhibitory activity against elastase, was identified as Phenolic substance using various colorimetric methods, UV, and IR. $IC_{50}$/ values of phenolic substance purified from Areca catechu were 26.9 $\mu\textrm{g}$/$m\ell$ for porcine pancreatic elastase (PPE) and 60.8 $\mu\textrm{g}$/$m\ell$ for human neutrophil elastase (HNE). This Phenolic substance showed more potent activity than those of reference compounds, oleanolic acid (76.5 $\mu\textrm{g}$/$m\ell$ for PPE, 219.2 $\mu\textrm{g}$/$m\ell$ for HNE) and ursolic acid (31.0 $\mu\textrm{g}$/$m\ell$ for PPE, 118.6 $\mu\textrm{g}$/$m\ell$ for HNE). According to the Lineweaver-Burk Plots, the inhibition against both PPE and HNE by this phenolic substance was competitive with substrate. Phenolic substance from Areca catechu exhibited high free radical scavenging effect ($SC_{50}$/ : 6 $\mu\textrm{g}$/$m\ell$) and inhibited effectively hyaluronidase activity ($IC_{50}$/: 210 $\mu\textrm{g}$/$m\ell$). These results suggest that the Phenolic substance Purified from Areca catechu showed anti-aging effect by protecting connective tissue proteins.

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