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Purification and Characterization of Extracellular Lipase from Streptomyces coelicolor A3(2) (Streptomyces coelicolor A3(2)로 부터 세포외 lipase의 정제와 특성)

  • Shim, Moon-soo;Kim, Jae-heon
    • Korean Journal of Microbiology
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    • v.33 no.4
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    • pp.237-241
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    • 1997
  • Lipase (EC 3.1.1.3) in the culture filtrate of Streptomyces coelicolor A3(2) was active on ${\alpha}$-naphthyl-butyrate as well as on various triacylglycerols with different lengths of acyl chains. The extracellular lipase was purified 15-fold by ammonium sulfate fractionation, Sephadex G-100, DEAE-Cellulose and Phenyl-Sepharose CL4B column chromatography with overall yield of 16%. It showed an molecular weight of 34.7 kDa by SDS-polyacrylamide gel electrophoresis. The enzyme activity with tributyrin as substrate was optimal at pH 8.0~9.0 and at $37^{\circ}C$. The enzyme activity decreased when the chain length of acyl group of triacyglycerol increased. A-factor, a hormone-like regulator of Streptomyces differentiation inhibited the lipase activity, which might corelate with the low enzyme activity in early exponential growth phase.

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Comparative Analysis and Evaluation of Dietary Intakes of Koreans by Age Groups: (5) Meal Patterns (서울 및 근교에 거주하는 한국인의 연령별 식생활 비교 및 평가: (5) 끼니별 섭취)

  • 심재은;백희영;문현경;김영옥
    • Journal of the Korean Home Economics Association
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    • v.42 no.8
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    • pp.169-185
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    • 2004
  • This study was performed to investigate the characteristics of meal patterns among Koreans of different age groups. The study subjects consisted of preschool children (PC), elementary school children (EC), high school students (HS), college students (CS), and adults over 30 years old living in Seoul and the surrounding areas. A dietary survey was conducted using 24-hour recall method from 1996-1998 and data were collected from 2392 subjects. The subjects over 30 years old were classified as 30∼49 years, 50∼69 years, and over 70 years. EC, HS, and CS groups consumed less than 20% of their total energy intake from breakfast. PC and EC consumed over 30% of their energy from snacks. In all age groups, the proportion of subjects who skipped a meal was highest for breakfast and lowest for dinner. Especially, 31.4% of the CS group skipped breakfast. Breakfast and snacks provided higher quality meals than the others when each meal was assessed by the index of nutritional quality (INQ). Especially, lunch of the HS group, each meal of the EC group and subjects over 70 years of age had a lower diet quality than the others did. The major snack items for PC, EC, HS, and CS groups were milk, ice cream, and snack cakes, but those for subjects over 30 years of age were fruits. The PC group obtained 35.5% of their energy from snacks, but the proportion of snack energy decreased with increasing age. PC and EC groups obtained a great proportion of calcium, vitamin B2, fat, and cholesterol intake from snacks. Snack foods provided 53.2%, 42.4%, 45.4%, and 24.6% of those nutrients, respectively, for the PC group and 54.2%, 43.5%, 37.4%, and 26.8%, respectively, for the EC group. These results suggested that each age group had specific meal patterns. Especially, breakfast and snack intake seemed to complement overall diet quality. However, more intervention is needed to improve overall dietary nutrition, e.g., efforts to decrease fat and cholesterol proportion and increase breakfast consumption. These dietary characteristics among different age groups present foundation data for the development of methods to resolve the nutritional problems of each age group.

Desmutagenicity of Tea Extracts from Green Tea, Oolong Tea and Black Tea (녹차, 오룡차 및 홍차 추출물의 돌연변이원성 억제작용)

  • 김선봉;여생규;김인수;안철우;박영호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.1
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    • pp.160-168
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    • 1995
  • Desmutagenicities against 2-amino-1-methyl-6-phenylimidazo[4, 5-b] pyridine(PhIP) and 2-amino-3, 8-dimethylimidazo[4, 5-f]quinoxaline(MelQx) of tea extracts (steamed green tea, roasted green tea, oolong tea and black tea) were investigated. All the fractions obtained from tea extracts showed strong desmutagenic activity against PhIP and MeIQx toward S. typhimurium TA 98 in the presence of the S-9 mix. The crude catechin fraction exhibited the strongest desmutagenic activity. Among these tea extracts, black tea especially exhibited the strongest desmutagenic activity and the activity was 70.9~91.0% against PhIP and 92.2~98.8% against MelQx at a concentration(0.5~1.0mg/plate) for drinking. The activity of authentic catechins of (-)-EGC, (-)-EGCg, (-)-ECg and (-)-EC were 79.5%, 60.2%, 46.1% and 43.5% against PhIP, and were 52.3%, 11.6%, 8.2% and 22.1% against MelQx by addition of 1.0mg/plate, respectively. The desmutagenic activity was supposedly due to the (-)-EGCg, (-)-EGC and (-)-EC in tea polyphenols, and the browning materials. The desmutagenicity was stronger when mutagens were preincubated with S-9 mix after reaciton with black tea extracts than when preincubated with them after reaction with S-9 mix. The desmutagenicity of tea extracts was rather expressed by reacting directly with mutagens than by deactivating the activated forms of mutagens.

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NF-κB Activation and PPAR Transactivational Effects of a New Aliphatic Acid Amide from Pericarps of Zanthoxylum piperitum

  • Yang, Seo Young;Tai, Bui Huu;Song, Seok Bean;Li, Wei;Yan, Xi Tao;Sun, Ya Nan;Nguyen, Phuong Thao;Kim, Young Ho
    • Bulletin of the Korean Chemical Society
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    • v.35 no.8
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    • pp.2361-2366
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    • 2014
  • A new aliphatic acid amide, ZP-amide F (1), and eight known compounds, including bungeanumamide A (2), tumuramide C (3), ZP-amide A (4), ZP-amide B (5), ZP-amide D (6), hyperin (7), quercitrin (8), and (-)-sesamin (9), were isolated from pericarps of Zanthoxylum piperitum. The effects of these compounds on $TNF{\alpha}$-induced NF-${\kappa}B$ activation and transactivational activity of PPARs, including $PPAR{\alpha}$, $PPAR{\beta}({\delta})$ and $PPAR{\gamma}$ subtypes, were evaluated. Compounds 7 and 9 exhibited potent inhibitory effects on $TNF{\alpha}$-induced NF-${\kappa}B$ activation with $IC_{50}$ values of 5.50 and $8.10{\mu}M$, respectively. Aliphatic acid amide compounds 3, 4 and 6 displayed enhanced effects on PPAR transactivational activity with $EC_{50}$ values of 47.12, 19.13 and $12.02{\mu}M$, respectively. Among them, compound 4 demonstrated an increase in $PPAR{\alpha}$ transactivational activity, compound 3 showed a moderate increase on all PPAR subtypes, whereas compound 6 displayed weak PPAR transactivational activity.

Evaluation of Winter Green Manure Crops as Alternative of Expeller Cake Fertilizer on Environment-friendly Red Pepper Production (친환경 고추 재배에서 유박 비료 대체재로서 동계 풋거름 작물의 이용 가능성 평가)

  • Choi, Jang-Yong;Yun, Yeo-Uk;Lee, Jin-Il;Hong, Ki-Heung;Kang, Young-Sik
    • Korean Journal of Organic Agriculture
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    • v.30 no.2
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    • pp.279-290
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    • 2022
  • This study was conducted to find out that winter green manure crops could be efficient replacements of expeller cake fertilizers, which were mostly imported, on environment-friendly red pepper production. Four treatments were compared under the condition of plastic film house: 1) barley (B); 2) hairy vetch (HV); 3) mixtures of barley and hairy vetch (B/HV); 4) expeller cake (EC). Total nitrogen content in hariy vetch was 3.6%, which was higher than 1.5~1.8% in barley and mixtures of barley and hairy vetch. P2O5 and K2O contents were similar in all green manures. Supplying amount of nitrogen from B, HV and B/HV plot, which were 172 kg ha-1, 193 kg ha-1, and 198 kg ha-1, were higher than amount of basal nitrogen required by soil testing but were lower than that of total nitrogen, respectively. Among the green manure crops, C/N ratio of HV was the lowest at 11.8, showing a similar value to that of expeller cake, but that of B was the highest at 30.6. Total yield was no significant difference in all treatment plots although B/HV plot showed the highest yield, but initial yield in B plot with high C/N ratio was lower than that in EC plot.

Purification and Properties of Aspergillus 3cuum exoinulinase (Aspergillus ficuum 조효소액으로부터 Exoinulinase의 정제 및 특성)

  • 한상배;송근섭;유향숙;노민환;이태규;손희숙;우순자;엄태봉
    • Microbiology and Biotechnology Letters
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    • v.19 no.3
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    • pp.253-258
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    • 1991
  • - An exoinulinase (EC 3.2.1.80) was purified from a commercial inulinase preparation from Aspergillus ficuum using ion exchange chromatography on CM-Sephadex C-50 and DEAESepharose 6B and HPLC gel filtration on a Protein Pak 125 column. Native exoinulinase had a molecular weight of 83, 000$\pm$ 1, 000 and was glycoprotein. Optimal pHs of the enzyme were ranged from 4.4 to 4.7. About ninety five percent of the whole activity was maintained even after incubation of 8 hours at $55^{\circ}C$.The enzyme was a typical non-specific P-fructofuranosidase, of which I/S ratio appears to be 0.35.

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Metabolic Adjustments of Lactate Dehydrogenase Isozymes to the Environmental Temperature in Bluegill (Lepomis macrochirus) (환경온도에 대한 파랑볼우럭(Lepomis macrochirus) 젖산탈수소효소 동위효소들의 대사조절)

  • Ku, Bora;Cho, Sung Kyu;Yum, Jung Joo
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1105-1112
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    • 2016
  • The aim of this study was to examine the metabolic adjustment of lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes to the environmental temperature in bluegill (Lepomis macrochirus). This study included three groups of bluegill collected in April (group Ⅰ), May (group Ⅱ), and September (group Ⅲ). The LDH activities of skeletal muscle, heart, and brain tissues were higher in group Ⅲ than in groups Ⅰ and Ⅱ. The citrate synthase (EC 4.1.3.7, CS) activity was higher in skeletal muscle but lower in heart and brain tissues of group Ⅱ as compared to group Ⅰ. In contrast, the CS activity was lower in skeletal muscle and higher in heart and brain tissues in group Ⅲ than in group Ⅱ. Furthermore, the LDH/CS activity ratio was higher in the skeletal muscle and brain in group Ⅲ than in groups Ⅰ and Ⅱ. Accordingly, anaerobic metabolism was increased in group Ⅲ. LDH A4, A2B2, and B4 isozymes were expressed in skeletal muscle, heart, liver, and brain tissues. The LDH C hybrid was detected in brain tissue. The LDH A4 isozyme was successfully purified by affinity chromatography. The molecular weight of the purified LDH A4 isozyme was 136 kDa and its optimal pH for enzymatic activity was 8.0. The KmPYR values of LDH in skeletal muscle were 0.161-0.227 mM using pyruvate as a substrate. These kinetic properties of LDH in skeletal muscle are consistent with the fact that bluegill is a cold-adapted species. These results may be useful for predicting the habitat use of this fish.

Thelephoric acid and Kynapcin-9 in Mushroom Polyozellus multiflex Inhibit Prolyl Endopeptidase In Vitro

  • Kwak, Ju-Yeon;Rhee, In-Koo;Lee, Kyung-Bok;Hwang, Ji-Sook;Yoo, Ick-Dong;Song, Kyung-Sik
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.798-803
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    • 1999
  • Prolyl endopeptidase [PEP; EC 3.4.21.26], a serine protease which is known to cleave peptide bonds on the carboxy side of a proline residue, plays an important role in the degradation of proline-containing neuropeptides that have been suggested to participate in learning and memory processes. An abnormal increase in the level of PEP, which can lead to generation of $A{\beta}$, is also suggested to be involved in Alzheimer's type senile dementia. In the course of screening PEP inhibitors from Basidiomycetes, the mushroom Polyozellus multiplex exhibited a high inhibitory activity against PEP. Two active compounds were isolated from the ethyl acetate soluble fraction by consecutive purification, using silica gel, Sephadex LH-20, and Lobar RP-18 chromatography. The chemical structures of these compounds were identified as thelephoric acid and 12-acety1-2,3,7,8-tetrahydroxy-[12H]-12-hydroxymethylbenzobis[I.2b,3.4b'] benzofuran-11-one (kynapcin-9) by spectral data including UV, IR, MS, HR-MS, $^1H-,{\;}^{13}C-$, and 2D-NMR. The $IC_{50}$ values of the thelephoric acid and kynapcin-9 were 0.157 ppm (446nM) and 0.087 ppm (212nM) and their inhibitor constants ($K_i$) were 0.73ppm ($2.09{\;}\mu\textrm{m}$) and 0.060 ppm (146 nM), respectively. Furthermore, they were non-competitive with a substrate in Dixon plots.

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Anti-Inflammatory and PPAR Transactivational Effects of Oleanane-Type Triterpenoid Saponins from the Roots of Pulsatilla koreana

  • Li, Wei;Yan, Xi Tao;Sun, Ya Nan;Ngan, Thi Thanh;Shim, Sang Hee;Kim, Young Ho
    • Biomolecules & Therapeutics
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    • v.22 no.4
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    • pp.334-340
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    • 2014
  • In this study, 23 oleanane-type triterpenoid saponins were isolated from a methanol extract of the roots of Pulsatilla koreana. The NF-${\kappa}B$ inhibitory activity of the isolated compounds was measured in $TNF{\alpha}$-treated HepG2 cells using a luciferase reporter system. Compounds 19-23 inhibited $TNF{\alpha}$-stimulated NF-${\kappa}B$ activation in a dose-dependent manner, with $IC_{50}$ values ranging from $0.75-8.30{\mu}M$. Compounds 19 and 20 also inhibited the $TNF{\alpha}$-induced expression of iNOS and ICAM-1 mRNA. Moreover, effect of the isolated compounds on PPARs transcriptional activity was assessed. Compounds 7-11 and 19-23 activated PPARs the transcriptional activity significantly in a dose-dependent manner, with $EC_{50}$ values ranging from $0.9-10.8{\mu}M$. These results suggest the presence of potent anti-inflammatory components in P. koreana, and will facilitate the development of novel anti-inflammatory agents.

Synthesis and Antiviral Activity of Novel 2′-Methyl and 4′-Phenyl Branched Carbocyclic Nucleosides (2′-메칠 및 4′-페닐 측쇄를 가진 새로운 카보사이클릭 뉴크레오사이드의 합성 및 항바이러스 약효검색)

  • 양선화;홍준희
    • YAKHAK HOEJI
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    • v.48 no.1
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    • pp.88-92
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    • 2004
  • In this study; a series of 2',4'-doubly branched carbocyclic nucleosides (8,9,10) were synthesized from simple acyclic ketone derivative as starting material. The installation of the 4'-quaternary carbon needed was carried out using a 〔3,3〕-sigmatropic rearrangement. In addition, the introduction of a methyl group in the 2'-position was accomplished by Grig-nard reaction. Bis-vinyl was successfully cyclized using a Grubbs' catalyst II. The natural bases (adenine, cytosine, uracil) were efficiently coupled with the use of a Pd(0) catalyst. Although all the synthesized compounds were assayed against several viruses, only cytosine analogue 9 showed weak antiviral viral activity (EC$_{50}$=45.4 $\mu$M) against CoxB3 virus.s.