• Title/Summary/Keyword: Anion-exchange

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Purification and Characterization of Polyphenol Oxidase from Flammulina velutipes (팽나무버섯 polyphenol oxidase의 정제 및 특성)

  • Pyo, Han-Jong;Son, Dae-Yeul;Lee, Chan
    • Korean Journal of Food Science and Technology
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    • v.34 no.4
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    • pp.552-558
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    • 2002
  • Polyphenol oxidase from Flammulina velutipes was purified and characterized. Purification of polyphenol oxidase was achieved by ammonium sulfate precipitation, Superdex G-200 gel filtration chromatography, Phenyl superose affinity chromatography, Mono-Q anion exchange chromatography and Superdex S-200 gel filtration chromatography on FPLC. After these purification steps specific activity of purified polyphenol oxidase increased to 199.1 units/mg. Polyphenol oxidase from F. velutipes was composed of a single polypeptide with molecular weight of about 40 kDa. Optimum pH and temperature for the enzyme reaction were found to be 6.0 and $25^{\circ}C$, respectively. The activity of the enzyme gradually decreased at acidic pH between 3 and 5, and the enzyme lost its activity at alkaline pH between 8 and 10. This enzyme exhibited high substrate specificity to o-diphenols. Km-values for L-DOPA and caffeic acid were found to be 3.97 mM and 1.78 mM, respectively. 2-mercaptoethanol, L-ascorbic acid, sodium bisulfite, EDTA and $Mg^{2+}$ inhibited the activity of pholyphenol oxidase and $Cu^{2+}$, $Fe^{2+}$, $Zn^{2+}$ and $Ni^{2+}$ increased enzyme activity. The activity of enzyme was well maintained at $-70^{\circ}C$ for over 4 months, and at $-20^{\circ}C$ for 1 months.

Immunostimulatory activity and structural characteristics of neutral polysaccharides isolated from ginseng leaves fermented by Cordyceps sinensis (동충하초균으로 발효한 인삼잎에서 분리한 중성다당의 면역활성 및 구조적 특성)

  • Cha, Ha Young;Park, Hye-Ryung;Shin, Kwang-Soon
    • Korean Journal of Food Science and Technology
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    • v.53 no.5
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    • pp.570-577
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    • 2021
  • This study elucidated the biological activities and structural characteristics of polysaccharides isolated from ginseng leaves fermented using Cordyceps sinensis (GLF). GLF comprised at least 18 glycosyl linkages, including 4-linked glucose residues (24.0%). To characterize the neutral polysaccharides in GLF, it was further fractionated by anion exchange chromatography, and the unabsorbed fraction (GLF1) was isolated. Peritoneal macrophages stimulated with GLF1 produced various cytokines in a dose-dependent manner. The properties and activities of the four subfractions (PHI, PHIA1-PHIA3) obtained after sequential enzymatic digestion were examined. PHI and PHIA3 primarily comprised glucose, whereas PHI exhibited an iodine-color reaction. Furthermore, the PHIA1-3 fractions indicated that cytokine production was completely inhibited. These results suggest at the immune activities of GLF1 may be due to the α-(1→4)-glucan branched at the C(O)6 position, which was produced by C. sinensis.

Comparison of Volatile Fatty Acids, Monosaccharide Analysis and Metabolic Profiling in Rumen Fluid According to Feeding Methods (사료 급여 방식에 따른 반추위액 내 휘발성지방산과 단당류 비교 분석과 대사산물 분석)

  • Eom, Jun-Sik;Lee, Shin-Ja;Lee, Yoo-Gyung;Lee, Sung-Sill
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.12
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    • pp.814-824
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    • 2018
  • This study was conducted to investigate VFA, monosaccharides and metabolites in rumen fluid according to feeding methods. Three castrated Hanwoo steers were used to the $3{\times}3$ Latin square design, 10 day for the diet adaptation period. VFA and monosaccharides which were not detected by HPLC and HPAEC however, those were detected by $^1H-NMR$. Among the metabolites measured by $^1H-NMR$ carbohydrate metabolites, pyruvate was detected only in the rumen fluid before feeding and succinate was detected before and after feeding rumen fluid. In amino acid total 9 metabolites were detected. In lipid metabolites, ethylene glycol was significantly higher (P<0.05) in before feeding Con group. In aliphatic acylic metabolite, trimethylamine N-oxide was no significant difference observed compare to Con group. In this study, many metabolites were observed in the rumen fluid by $^1H-NMR$, and it confirmed that rumen metabolic products were changed by feeding methods.

Phytochemical analysis of Panax species: a review

  • Yang, Yuangui;Ju, Zhengcai;Yang, Yingbo;Zhang, Yanhai;Yang, Li;Wang, Zhengtao
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.1-21
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    • 2021
  • Panax species have gained numerous attentions because of their various biological effects on cardiovascular, kidney, reproductive diseases known for a long time. Recently, advanced analytical methods including thin layer chromatography, high-performance thin layer chromatography, gas chromatography, high-performance liquid chromatography, ultra-high performance liquid chromatography with tandem ultraviolet, diode array detector, evaporative light scattering detector, and mass detector, two-dimensional high-performance liquid chromatography, high speed counter-current chromatography, high speed centrifugal partition chromatography, micellar electrokinetic chromatography, high-performance anion-exchange chromatography, ambient ionization mass spectrometry, molecularly imprinted polymer, enzyme immunoassay, 1H-NMR, and infrared spectroscopy have been used to identify and evaluate chemical constituents in Panax species. Moreover, Soxhlet extraction, heat reflux extraction, ultrasonic extraction, solid phase extraction, microwave-assisted extraction, pressurized liquid extraction, enzyme-assisted extraction, acceleration solvent extraction, matrix solid phase dispersion extraction, and pulsed electric field are discussed. In this review, a total of 219 articles published from 1980 to 2018 are investigated. Panax species including P. notoginseng, P. quinquefolius, sand P. ginseng in the raw and processed forms from different parts, geographical origins, and growing times are studied. Furthermore, the potential biomarkers are screened through the previous articles. It is expected that the review can provide a fundamental for further studies.

Changes in isomaltooligosaccharides during fermentation of Makgeolli (막걸리 발효 중 이소말토올리고당의 변화)

  • Noh, Dong-Hyun;Jeong, Seok-Tae;Park, Boram;Kim, Yong-suk;Lim, Bora
    • Korean Journal of Food Science and Technology
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    • v.54 no.3
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    • pp.351-361
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    • 2022
  • This study aimed to investigate the differences in the isomaltooligosaccharides present during Makgeolli production according to the type of Nuruk used, fermentation period, and presence of microorganisms. Makgeolli was fermented for 15 days using three kinds of Nuruk (Soyul, Sansung, and Jinju) with and without microbial growth inhibitors. Isomaltooligosaccharide contents were analyzed using high performance anion exchange chromatography. The most abundant isomaltooligosaccharide was panose, which was highest in Makgeolli produced using Soyul Nuruk (SH) on day 6 (24.7 mM), followed by Makgeolli prepared using Sansung Nuruk (SS) on day 2 (18.2 mM) and Makgeolli prepared using Jinju Nuruk (JJ) on day 3 (10.8 mM). Isomaltotriose and isomaltotetraose, which were generated in the control, were not detected when microbial growth was suppressed. Based on these results, isomaltooligosaccharide production is affected more by the enzymes produced by microorganisms during Makgeolli fermentation than by Nuruk itself.

Effect of Water Soluble fraction from Japanese Larch Wood on Sawdust Cultivation of Lentinula edodes (일본잎갈나무재의 수용성추출물 첨가가 표고버섯의 톱밥재배에 미치는 영향)

  • Cho, Nam-Seok;Chung, Hung-Chae;Kim, Dong-Hun;Lee, Sang-Sun;Ohga, Shoji;Leonowicz, A.
    • Journal of the Korean Wood Science and Technology
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    • v.32 no.1
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    • pp.35-44
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    • 2004
  • The water soluble fractions(WSF) from Japanese larch wood were isolated, purified by anion exchange resin and Sephadex gel filtration and identified its chemical structure by means of periodate oxidation and methylation reactions. Its major components are arabinose and galactose (1 : 3.4). Based on the results of periodate oxidation, methylation and gas chromatographic analysis of purified WSF, main chain is composed of β-1,3-glycosidic linkage among D-galactopyranoses, and two different side chains; β-1,6-glycosidic linkage among 2-3 units of D-galactopyranoses and β-1,6-glycosidic linkage between 1-2 units of D-galactopyranose and L-arabinopyranose. Addition of WSF to culture media of oak mushroom (Lentinula edodes) accelerated the mycelial growth. In the case of PDA cultures, 2 percent addition of WSF in Sanlim No. 6 strain and 4 percent of WSF in Mok-H strain mostly enhanced the mycelial growth of the mushroom. In the case of sawdust cultures, 4 percent addition of WSF in two strains showed the best mycelial growth. High percentages addition of WSF inhibited mycelial growth of the mushroom. Mushroom production was increased with addition of WSF. By the addition of WSF, ergosterol contents in the media were quite high at the colonized stage and rapidly increased at the fruiting stage. Therefore the ergosterol content could be utilized as an indicator to evaluate the culture maturity for the mushroom fruiting.

Improved Copper Ion Recovery Efficiency through Surface Modification of Membranes in the Electrodialysis/Solvent Extraction Process (전기투석/용매추출 공정에서 멤브레인 표면 개질을 통한 구리 이온의 회수 효율 향상)

  • Joongwon, Park;Rina, Kim;Hyunju, Lee;Min-seuk, Kim;Hiesang, Sohn
    • Membrane Journal
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    • v.32 no.6
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    • pp.486-495
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    • 2022
  • This study presents the improved recovery efficiency of rare metal ions through the modified separation membrane wettability and hydrogen ion permeation in the anion exchange membrane (AEM) under the recovery process of combined electrodialysis and solvent extraction. Specifically, the wettability of the separator was enhanced by hydrophilic modification on one separator surface through polydopamine (PDA) and lipophilic modification on the other surface through SiO2 or graphene oxide (GO). In addition, the modified surface of AEM with polyethyleneimine (PEI), PDA, poly(vinylidene fluoride) (PVDF), etc. reduces the water uptake and modify the pore structure for proton ions generation. The suppressed transport resulted in the reduced hydrogen ion permeation. In the characterization, the surface morphology, chemical properties and composition of membrane or AEM were analyzed with Scanning Electron Microscopy (SEM) and Fourier Transform-Infrared Spectroscopy (FT-IR). Based on the analyses, improved extraction and stripping and hydrogen ion transport inhibition were demonstrated for the copper ion recovery system.

Purification and Characterization of Mitochondrial Mg2+-Independent Sphingomyelinase from Rat Brain

  • Jong Min Choi;Yongwei Piao;Kyong Hoon Ahn;Seok Kyun Kim;Jong Hoon Won;Jae Hong Lee;Ji Min Jang;In Chul Shin;Zhicheng Fu;Sung Yun Jung;Eui Man Jeong;Dae Kyong Kim
    • Molecules and Cells
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    • v.46 no.9
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    • pp.545-557
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    • 2023
  • Sphingomyelinase (SMase) catalyzes ceramide production from sphingomyelin. Ceramides are critical in cellular responses such as apoptosis. They enhance mitochondrial outer membrane permeabilization (MOMP) through self-assembly in the mitochondrial outer membrane to form channels that release cytochrome c from intermembrane space (IMS) into the cytosol, triggering caspase-9 activation. However, the SMase involved in MOMP is yet to be identified. Here, we identified a mitochondrial Mg2+-independent SMase (mt-iSMase) from rat brain, which was purified 6,130-fold using a Percoll gradient, pulled down with biotinylated sphingomyelin, and subjected to Mono Q anion exchange. A single peak of mt-iSMase activity was eluted at a molecular mass of approximately 65 kDa using Superose 6 gel filtration. The purified enzyme showed optimal activity at pH of 6.5 and was inhibited by dithiothreitol and Mg2+, Mn2+, Ni2+, Cu2+, Zn2+, Fe2+, and Fe3+ ions. It was also inhibited by GW4869, which is a non-competitive inhibitor of Mg2+-dependent neutral SMase 2 (encoded by SMPD3), that protects against cytochrome c release-mediated cell death. Subfractionation experiments showed that mt-iSMase localizes in the IMS of the mitochondria, implying that mt-iSMase may play a critical role in generating ceramides for MOMP, cytochrome c release, and apoptosis. These data suggest that the purified enzyme in this study is a novel SMase.

Characterization and Purification of Subtilosin A Produced by Bacillus vallismortis MCBL 1012 Isolated from Seasoned Dried Radish

  • Se-Yeon Lee;Dae-Ook Kang
    • Journal of Life Science
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    • v.34 no.8
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    • pp.576-587
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    • 2024
  • In this study, diverse bacterial strains were isolated from fermented foods to screen those with antibacterial activity. Among them, one strain, identified as Bacillus vallismortis MCBL 1012 through 16S rRNA gene sequence analysis, was selected for its bacteriocin production. The culture supernatant of B. vallismortis MCBL 1012 showed antibacterial activity, mainly against Gram-positive bacteria. Scanning electron microscopy (SEM) revealed that bacteriocin treatment led to cellular content leakages in Listeria monocytogenes KCCM 40307, Enterococcus faecium KCCM 12118, and Streptococcus mutans KCTC 3065. PCR analysis confirmed B. vallismortis MCBL 1012 harbored subtilosin A gene (sbo A). Antibacterial activity was decreased by proteolytic enzymes like proteinase K, subtilisin A, and α-chymotrypsin. The bacteriocin demonstrated stability at 40℃ and 60℃ for 120 min, and up to 80℃ for 60 min, with rapid activity loss at 100℃. It retained full antibacterial activity within a pH range of 4.0 to 8.0 and was not affected by up to 100% organic solvents like ethanol, methanol, acetonitrile, and tetrahydrofuran. Nevertheless, activity decreased with more than 40% isopropanol and 80% acetone. Most tested inorganic salts and detergents had no effect on antibacterial activity except, CuSO4 and NiSO4 at specified concentrations. The bacteriocin exerted its antibacterial effect through bactericidal action against L. monocytogenes KCCM 40307. The bacteriocin was purified by ammonium sulfate precipitation, DEAE anion exchange chromatography, and RP-HPLC. The purification resulted in a final yield of 0.03% and a 283.7-fold increase in specific activity. MALDI-TOF MS analysis determined the exact molecular weight of purified bacteriocin to be 3,326.1 Da.

A Study on the Recycling Process of Nickel Recovery from Inconel 713C Scrap based on Hydrometallurgy (인코넬 713C 스크랩으로부터 니켈 자원 회수를 위한 습식제련 기반 재활용공정 연구)

  • Min-seuk Kim;Rina Kim;Kyeong-woo Chung;Jong-Gwan Ahn
    • Resources Recycling
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    • v.33 no.4
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    • pp.36-46
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    • 2024
  • We investigated a hydrometallurgical process of nickel recovery from Inconel 713C scrap. The process proceeded with a series of i) comminution of pyrometallurgical treated scrap, ii) sulfuric acid leaching, iii) solvent extraction of unreacted acid, molybdenum, aluminum, and precipitation of chromium, iv) crystallization of nickel sulfate by vacuum evaporation, and v) nickel electrowinning. The nickel-aluminum intermetallic compound, Ni2Al3, was formed by the pyrometallurgical pretreatment readily grounded under 75 ㎛. Sulfuric acid leaching was done for 2 hours in 2 mol/L, 20 g/L solid/liquid ratio, and 80 ℃. It revealed that over 98 % of nickel and aluminum was dissolved, whereas 28 % of molybdenum was. A nickel sulfate solution with 2.34 g/L for the crystallization of nickel sulfate hydrate was prepared via solvent extraction and precipitation. Over 99 % of molybdenum and aluminum and 93 % of chromium was removed. Nickel metal with 99.9 % purity was obtained by electrowinning with the nickel sulfate monohydrate in the cell equipped with anion exchange membranes for catholyte pH control. The membrane did not work well, resulting in a low current efficiency of 73.3 %.