• Title/Summary/Keyword: soluble/total fraction

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Fractionation of enzymatically methylated acid-insoluble proteins from thymus nuclei

  • Lee, Hyang-Woo;Kim, Sang-Duk;Paik, Woon-Ki
    • Archives of Pharmacal Research
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    • v.9 no.3
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    • pp.157-161
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    • 1986
  • Isolated calf thymus nuclei were in vitro methylated with S- adenosy-L-methyl-$^{14}C$ methionine, and the proteins were fractionated according to their solubilities. Histone fraction ($H_{2}SO_{4}$-soluble fraction) contained approximately 60% total radioactivity incorporated, while "residual protein" which was ($H_{2}SO_{4}$-insoluble contained the remaining radio-activity. The "residual protein" was further fractionated into various acidic proteins, which contained very littel of the radioactivity. However, the protein fraction eluted from DEAE-cellulose with 0.5 N NaOH contained the largest amount of radioactivity. This protein was found to be basic in nature by amino analysis.

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Protein Fractionation of Whole Crop Silages, and Effect of Borate-phosphate Buffer Extraction on In vitro Fermentation Characteristics, Gas Production and Degradation (사료작물 사일리지의 단백질 분획 및 Borate-phosphate Buffer 추출이 In vitro 발효성상, Gas 발생 그리고 분해율에 미치는 효과)

  • Shinekhuu, Judder;Jin, Guang-Lin;Ji, Byung-Ju;Li, Xiangzi;Oh, Young-Kyoon;Hong, Seong-Ku;Song, Man-Kang
    • Journal of Animal Science and Technology
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    • v.51 no.5
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    • pp.369-378
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    • 2009
  • Protein fractionation was evaluated from whole crop silages of rye (RS), wheat (WS), triticale (TS), oat (OS), barley (BS), and rice straw silage (RSS), and in vitro trial was carried out to examine the effect of silage and extraction of soluble protein on fermentation characteristics, total gas production and degradation. Soluble protein of silages was extracted with borate-phosphate buffer, and fermentation characteristics, gas production and degradation of silages were estimated by incubating anaerobically the mixed solution of strained rumen fluid and artificial saliva (1:1, v/v) containing dried and ground silages placed in nylon bag at $39^{\circ}C$ up to 48h. Soluble protein (SP) content was lowest for RSS as 2.11% in total CP compared to those for other silages. Highest A fraction (NPN) was observed from RS (74.33% of total CP) while those from TS and RSS were relatively low (48%). B2 fraction was relatively higher for RS, RSS and WS than for TS and BS. $B_3$ fraction was lowest in WS among silages. C fraction (27.07) in RSS was higher than in other silages (1.40~9.93%). pH in incubation solution was increased (P<0.01~P<0.001) for extracted silages up to 12h but decreased (P<0.01) at 48h for non-extracted ones. Contents of ammonia-N (P<0.001) and total VFA (P<0.01~P<0.001) were higher for non-extracted silages than for extracted ones. Acetate proportion was increased (P<0.001) in buffer extracted silages while those of propionate and butyrate were decreased (P<0.001) up to 24h incubation. Increased (P<0.001) total gas production was obtained from non-extracted silages up to 12h while gas production was increased (P<0.01) in extracted ones thereafter. In vitro degradation of dry matter and CP was increased (P<0.001) in non-extracted silages but that of neutral detergent fiber was increased (P<0.001) in extracted ones without difference among silages. Difference in mean values of degradability for each silage prior to- and post extraction with borate buffer, however, was not found among silages. It may be concluded that high NPN content of silages may reduce the protein availability in silages and borate buffer soluble components in silages can stimulate the early stage of fermentation.

Simplified the Screening and In Vitro Appraisal of Antioxidant, Cytotoxic, Thrombolytic, Antimicrobial and Membrane Stabilizing Activities of Lablab Purpures at a Time

  • Rahman, M. Saifur;Uddin, M. Gias;Alam, M. Badrul;Yoo, Jin Cheol
    • Journal of Integrative Natural Science
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    • v.7 no.3
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    • pp.173-182
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    • 2014
  • To simplify the different biological investigation of the methanolic extract and solvent-solvent partitioning of Lablab purpures (L. purpures) bark. In-vitro anti-oxidant study was determined using total DPPH radical scavenging assay. In vitro antimicrobial study was measured by observing zone of inhibition. The cytotoxic activity was studied using brine shrimp lethality bioassay and thrombolytic activity by clot disruption method. The antioxidant potential was evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and Folin-Ciocalteau reagents using butylated hydroxytolune (BHT) and ascorbic acid as standards. The Aqueous soluble fraction revealed the highest free radical scavenging activity ($IC_{50}=48.76{\mu}g/mL$). The antimicrobial screening of the bark of L. purpures exhibited mild to moderate activity in test microorganisms. The CSF showed the maximum relative percentage inhibition against Salmonella parathyphi (34.2%) for bacteria and C. albicans (28.8%) for fungi whereas, lowest relative percentage inhibition against Sarcina lutea (22.0%) for bacteria and Aspergillus niger (24.4%) for fungi. In the brine shrimp lethality bioassay, The $LC_{50}$ values of Carbon tetrachloride and N-Hexane soluble fraction were found $92.18{\mu}g/mL$, and $68.95{\mu}g/mL$ respectively while the $LC_{50}$ values of standard Vincristine sulphate was $1.37{\mu}g/mL$. The methanolic extract and its organic soluble fractions of Lablab purpureus at concentration 2.0 mg/mL, significantly protected the lysis of erythrocyte membrane induced by hypotonic solution and heat as compared to the standard, acetyl salicylic acid (0.10 mg/mL). The MSF and AQSF produced 61.48 % and 53.75% inhibition of hemolysis of RBC caused by hypotonic solution respectively, whereas acetyl salicylic acid (0.10 mg/mL) showed 76.42%. Ethanol extract of L. purpures and all of its different partitions exhibited moderate thrombolytic activity of 37.25%-2.40%. Very good preliminary screening and simplified experiments were able to show the different biological activity of methanolic extract and its soluble fractions of L. purpures at a time.

Fate of 14C - Carobofuran in Rice Plant and Paddy Soil (수도체(水稻體) 및 담수토양중(湛水土壤中) $^{14}C$-Carbofuran의 행적(行跡)에 관한 연구(硏究))

  • Lee, Young-Deuk;Lee, Kyung-Hwi;Park, Chang-Kyu
    • Korean Journal of Environmental Agriculture
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    • v.6 no.1
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    • pp.7-16
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    • 1987
  • A study was undertaken to investigate the absorption, translocation and metabolism of carbofuran in rice paddies. Ring-3-$^{14}C-carbofuran$ applied onto the paddy soil surface was rapidly absorbed and translocated into rice plants. Within 2 days after treatment, it was observed that carbofuran reached shoot tips and accumulated. More than 15% of total radioactivity was recovered in rice plant from 3 to 20 days after treatment. In organic soluble fraction of rice plant extract, 3-hydroxycarbofuran was the major metabolite recording 43% and 4% of total organic soluble radioactivity in shoot and root at 20 days respectively. 3-Ketocarbofuran and phenolic metabolites including carbofuran phenol, 3-hydroxycarbofuran phenol and 3-ketocarbofuran phenol were also detected in the organic soluble fractions. Some glycosidic conjugates of carbofuran metabolites were found in water soluble fraction of rice plant extract and 3-hydroxycarbofuran was the most abundant aglycone. Radioactivity in paddy soil was rapidly decreased until 3 days after treatment and then maintained almost constant level. A significant portion (42∼56 %) of the total radioactivity remained in soil as nonextractable residue from 5 to 20 days after treatment. The nonextractable radioactivity was mainly located in soil organic matter distributing in humin, fulvic acid and humic acid fractions with the decreasing order. Evolution of $^{14}CO_2$ from ring cleavage of $3-^{14}C-carbofuran$ was negligible recording only 1.8% of total radioactivity during 20 days after treatment.

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Antidiabetic Activity of Mori Folium Ethanol Soluble Fraction in db/db mice (db/db 마우스에서 상엽 에탄올가용분획의 항당뇨활성)

  • Ryu, Jeong-Wha;Seo, Seong-Hoon;Chung, Sung-Hyun
    • YAKHAK HOEJI
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    • v.42 no.6
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    • pp.613-620
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    • 1998
  • Antidiabetic activity of Mori folium ethanol soluble fraction (MFESF) was examined in db/db mice, which is a spontaneously hyperglycemic, hyperinsulinemic and obese animal model . 500 and 1000mg/kg dose for MFFSF (designated by SY 500 and SY 1000, respectively) and 5mg/kg dose for acarbose were administered for 6 weeks. Body weight gain, fasting and non-fasting serum glucose, glycated hemoglobin and triglyceride were all reduced dose dependently when compared between db/db control group and MFESF treated group. At 11th and 13th week after birth, MFESF increased an insulin secretion which may result in lowering serum glucose level. Total activities of sucrase and maltase in SY 500 treated group were decreased when compared to db/db control. On the other hand, those in SY 1000 and acarbose treated groups were increased. This result may suggest that proteins for sucrase and maltase were compensatorily induced due to significant inhibition of glycosidase-catalyzed reaction at doses administered in this study.

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Tyrosinase Inhibitory Phenolic Constituents of Smilax china Leaves (청미래덩굴 잎의 페놀성 성분 및 Tyrosinase 저해 활성)

  • Kim, Sang Hyun;Ahn, Jong Hoon;Jeong, Ji Yeon;Kim, Seon Beom;Jo, Yang Hee;Hwang, Bang Yeon;Lee, Mi Kyeong
    • Korean Journal of Pharmacognosy
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    • v.44 no.3
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    • pp.220-223
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    • 2013
  • In the course of screening tyrosinase inhibitory activity, total methanolic extract and EtOAc-soluble fraction of Smilax china leaves showed significant inhibitory activity. Further fractionation and isolation of the EtOAc-soluble fraction resulted in 12 phenolic compounds, which were identified as 4-hydroxybenzoic acid (1), 3,4-dihydroxybenzaldehyde (2), 3,4-dihydroxybenzoic acid (3), 3,4-dihydroxyacetophenone (4), 3-hydroxy-4-methoxy benzoic acid (5), trans-p-hydroxycinnamic acid (6), cis-p-hydroxycinnamic acid (7), trans-resveratrol (8), cis-resveratrol (9), dihydroresveratrol (10), moracin M (11) and kaempferol (12). Compounds 1-11 were first reported from this plant. Among the isolated compounds, compounds 2, 8, 9 and 12 showed strong inhibition on tyrosinase activity.

Effects of Heat Treatments on the Dietary Fiber Contents of Rice, Brown Rice, Yellow Soybean, and Black Soybean. (가열처리에 의한 백미, 현미, 노란콩 및 검정콩의 식이섬유 함량 변화)

  • 서우경;김영아
    • Korean journal of food and cookery science
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    • v.11 no.1
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    • pp.20-25
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    • 1995
  • The effects of cooking on the dietary fiber content in rice, brown rice, yellow soybean and black soybean were investigated. The dietary fiber contents were analyzed by Prosky's method(AOAC method) after boiling, microwave heating and autoclaving of the sample. It was showed that the different cooking methods resulted in different effects on the insoluble dietary fiber contents. Except yellow soybean, cooking time had little effect on insoluble dietary fiber contets in the other samples. The contents of soluble dietary fiber were generally increased by cooking. Increased cooking time reduced the content of soluble dietary fiber in brown rice but increased in rice. However, no significant differences caused by cooking time were observed for soluble dietary fiber in black soybean. The effects of cooking method on the total dietary fiber contents were similar to those of insoluble dietary fiber. The reasons for this might be that the main fraction of total dietary fiber was insoluble forms and the content of total dietary fiber was calculated as the sum of insoluble and soluble dietary fiber content.

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Difference of Leaching Liability of minerals in Forages (목초에 함유된 수종 무기물의 성분분획)

  • Sangdeog A. Kim;Shigekata Yoshida
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.11 no.1
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    • pp.12-16
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    • 1991
  • Three minerals in orchard grass (Dactylis glomerata Linn.) grown under different potassium (K20) levels in water culture, potassium(K), magnesium(Mg) and calcium(Ca) were divided into water, alcohol, and acid soluble fractions to know the reason of different lea¬ching rate. The results were as follows; (1) The percentage of K in each fraction to total K was not affected by the fertilization levels, and it was 90-93, 4-6, and 2~3% in water, methanol and 0.5% HCI soluble fractions, respectively. (2) About 800/c of the extractive Mg was water soluble, and 14-22% of total Mg were extracted by O. 5% HC! solution. (3) The percentage of water, O. 5% HCI- and 1 M HCI-soluble Ca to total Ca were affected by the fertilization levels, and were 26-46%, 44-54%, and 8-22%, respectively. From the above facts, it is known that the solubility in water was the highest in K and followed by Mg and Ca. While the solubility in HCI solution was opposite to that in water.

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Effect of Molecular Chaperones on the Soluble Expression of Alginate Lyase in E. coli

  • Shin, Eun-Jung;Park, So-Lim;Jeon, Sung-Jong;Lee, Jin-Woo;Kim, Young-Tae;Kim, Yeon-Hee;Nam, Soo-Wan
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.5
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    • pp.414-419
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    • 2006
  • When the alginate lyase gene (aly) from Pseudoalteromonas elyakovii was expressed in E. coli, most of the gene product was organized as aggregated insoluble particles known as inclusion bodies. To examine the effects of chaperones on soluble and nonaggregated form of alginate lyase in E. coli, we constructed plasm ids designed to permit the coexpression of aly and the DnaK/DnaJ/GrpE or GroEL/ES chaperones. The results indicate that coexpression of aly with the DnaK/DnaJ/GrpE chaperone together had a marked effect on the yield alginate lyase as a soluble and active form of the enzyme. It is speculated this result occurs through facilitation of the correct folding of the protein. The optimal concentration of L-arabinose required for the induction of the DnaK/DnaJ/GrpE chaperone was found to be 0.05mg/mL. An analysis of the protein bands on SDS-PAGE gel indicated that at least 37% of total alginate lyase was produced in the soluble fraction when the DnaK/DnaJ/GrpE chaperone was coexpressed.

A Bioactive Fraction from Streptomyces sp. Enhances Maize Tolerance against Drought Stress

  • Warrad, Mona;Hassan, Yasser M.;Mohamed, Mahmoud S.M.;Hagagy, Nashwa;Al-Maghrabi, Omar A.;Selim, Samy;Saleh, Ahmed M.;AbdElgawad, Hamada
    • Journal of Microbiology and Biotechnology
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    • v.30 no.8
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    • pp.1156-1168
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    • 2020
  • Drought stress is threatening the growth and productivity of many economical crops. Therefore, it is necessary to establish innovative and efficient approaches for improving crop growth and productivity. Here we investigated the potentials of the cell-free extract of Actinobacteria (Ac) isolated from a semi-arid habitat (Al-Jouf region, Saudi Arabia) to recover the reduction in maize growth and improve the physiological stress tolerance induced by drought. Three Ac isolates were screened for production of secondary metabolites, antioxidant and antimicrobial activities. The isolate Ac3 revealed the highest levels of flavonoids, antioxidant and antimicrobial activities in addition to having abilities to produce siderophores and phytohormones. Based on seed germination experiment, the selected bioactive fraction of Ac3 cell-free extract (F2.7, containing mainly isoquercetin), increased the growth and photosynthesis rate under drought stress. Moreover, F2.7 application significantly alleviated drought stress-induced increases in H2O2, lipid peroxidation (MDA) and protein oxidation (protein carbonyls). It also increased total antioxidant power and molecular antioxidant levels (total ascorbate, glutathione and tocopherols). F2.7 improved the primary metabolism of stressed maize plants; for example, it increased in several individuals of soluble carbohydrates, organic acids, amino acids, and fatty acids. Interestingly, to reduce stress impact, F2.7 accumulated some compatible solutes including total soluble sugars, sucrose and proline. Hence, this comprehensive assessment recommends the potentials of actinobacterial cell-free extract as an alternative ecofriendly approach to improve crop growth and quality under water deficit conditions.