• Title/Summary/Keyword: P10 partial protein

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Purification and Partial Characterization of a Lectin from the Fresh Leaves of Kalanchoe crenata (Andr.) Haw

  • Adenike, Kuku;Eretan, Oladiran Babalola
    • BMB Reports
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    • v.37 no.2
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    • pp.229-233
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    • 2004
  • A haemagglutinating protein from the saline extracts of Kalanchoe crenata leaves, which agglutinate all human blood types, was purified to homogeneity by ion-exchange chromatography on a DEAE-Cellulose column followed by gel filtration on a Sephadex G-100 column. The purified protein showed one band, both in non-denaturing PAGE and SDS-PAGE. The $M_r$ that was determined by SDS-PAGE was 44,000 Da and that estimated from gel filtration was 47,000. Treatment of the haemagglutinating protein with 5 mM EDTA diminished the haemagglutinating activity to 50% of the original level. The addition of divalent cations, 10 mM $Mg^{2+}$, 10 mM $Mn^{2+}$, or 10 mM $Ba^{2+}$, totally restored and enhanced the activity. The protein showed maximum activity over the 3-7 pH range and was heat-resistant. It was also a glycoprotein containing about 1.5% carbohydrate.

Utilization of fermented skipjack tuna viscera as a dietary protein source replacing fish meal or soybean meal for juvenile abalone Haliotis discus hannai

  • Lee, Sang-Min;Kim, Kyoung-Duck;Kim, Tae-Jin
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.73-73
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    • 2003
  • This study was carried out to evaluate the utilization of fermented skipjack tuna viscera (FSTV) in the diet for juvenile abalone Haliotis discus hannai. Lactobacillus bulgaricus was used for fermentation of skipjack tuna viscera. Eight isonitrogenous (about 30% crude protein) diets were formulated to include different levels (0%, 10%, 20% and 30%) of FSTV as a replacer of either dietary fish meal or soybean meal. Three replicate groups of abalone were fed the experimental diets containing different levels of FSTV for 7 weeks. The inclusion of FSTV up to 30% in fish meal-based diet had no significant effect on survival, body weight, shell growth, and proximate composition of abalone (P>0.05). Weight gain of abalone fed the diet substituting 10% FSTV for soybean meal was not significantly different to that of abalone fed the control diet, however this value decreased in abalone fed the 20% and 30% FSTV (P<0.05).The contents of crude protein and lipid of soft body in abalone fed soybean meal-based diets were significantly affected by dietary FSTV level (P<0.05). The results of this study indicate that FSTV can be used as a partial substitute protein source for fish meal or soybean meal in the formulated diet for juvenile abalone.

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Complementary DNA Cloning and Restriction Mapping of Nuclear Inclusion Body and Coat Protein Genes of Turnip Mosaic Virus-Ca Strain Genomic RNA (순무모자이크 바이러스 Ca계통 핵봉입체와 외피단백질 유전자의 cDNA 클로닝 및 제한효소 지도작성)

  • 류기현;박원목
    • Korean Journal Plant Pathology
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    • v.10 no.3
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    • pp.235-239
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    • 1994
  • Viral RNA was extracted from purified Chinese cabbage strain of turnip mosaic virus (TuMV-Ca) from infected leaves of turnip. Polyadenylated genomic viral RNA was recovered by oligo (dT) cellulose column chromatography and used as a template for the synthesis of complementary DNA (cDNA). Recombinant plasmids contained cDNA ranged from about 900 bp to 2, 450 bp were synthesized. Among the selected 41 transformants, pTUCA31 and pTUCA35 had over 2 Kbp cDNA insert. Restriction endonuclease patterns of the clones examined were very similar among them. Clones pTUCA23 and pTUCA31 were overlapped with pTUA35. The longest clone pTUCA35, encoding 3'-end, showed that it contained two sites for EcoRI, and one site for BamHI, ClaI, HincII, SacI and XbaI, respectively. The restriction mapping indicated that the clone pTUCA35 contained partial nuclear inclusion body gene, complete coding region of the coat protein and 3' untranslated region of TuMV-Ca genomic RNA.

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A Novel Anticoagulant Protein from Scapharca broughtonii

  • Jung, Won-Kyo;Je, Jae-Young;Kim, Hee-Ju;Kim, Se-Kwon
    • BMB Reports
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    • v.35 no.2
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    • pp.199-205
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    • 2002
  • An anticoagulant protein was purified from the edible portion of a blood ark shell, Scapharca broughtonii, by ammonium sulfate precipitation and column chromatography on DEAE-Sephadex A-50, Sephadex G-75, DEAE-Sephacel, and Biogel P-l00. In vitro assays with human plasma, the anticoagulant from 'S. broughtonii, prolonged the activated partial thromboplastin time (APTT) and inhibited the factor LX in the intrinsic pathway of the blood coagulation cascade. But, the fibrin plate assay did not show that the anticoagulant is a fibrinolytic protease. The molecular mass of the purified S. broughtonii anticoagulant was measured to be about 26.0kDa by gel filtration on a Sephadex G-75 column and SDS-PAGE under denaturing conditions. The optimum activity in the APTT assay was exhibited at pH 7.0-7.5 and $40-45^{\circ}C$ in the presence of $Ca^{2+}$.

Effect of Partial Freezing as a Means of Keeping Freshness II. Changes in Freshness and Gel Forming Ability of Conger Eel and Yellowtail during Storage by Partial Freezing (Partial Freezing에 의한 어육의 선도유지 효과에 대하여 2. Partial Freezing에 의한 붕장어 및 방어의 선도 및 어묵형성능의 변화)

  • LEE Yong-Woo;PARK Yeung-Ho;AHN Cheol-Woo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.1
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    • pp.27-35
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    • 1986
  • In succession to the previous paper, the present study was directed to investigate the effect of keeping freshness of conger eel (Astroconger myriaster) and yellowtail (Seriola quinqueradiata) by partial freezing, and the changes in the physical properties of fish meat paste product prepared with the muscle of conger eel during storage were also examined. The results obtained are summarized as follows: The period of keeping freshness (days in which k value reaches $20\%$) of conger eel and yellowtail by partial freezing was 10 days and 6 days, respectively. VBN content in the conger eel muscle showed 39.5 mg/100g by icing for 15 days, and did not show a great change by partial freezing and freezing, while that of yellowtail muscle reached at 32 mg/100g by icing, 20 mg/100g by partial freezing and 18 mg/100g by freezing for 15 days. The lipids extracted from the muscles of both fishes by icing were remarkably oxidized than those by partial freezing. The myofibrillar protein in the conger eel muscle during storage for 9 days decreased $3\%,\;10%\;and\;11\%$ by icing, partial freezing and freezing, respectively, and that of yellowtail muscle did $16\%,\;10%\;and\;4\%$ by icing, partial freezing and freezing, respectively. On the other hand, the alkali-soluble protein in both fishes increased with storage time. Gel strength of fish meat paste product prepared with the muscle of conger eel decreased to $35\%$ by icing, $74\%$ by partial freezing and $76\%$ by freezing for 10 days compared to control, and the expressible water increased 1.6 times, 1.2 times and 1.1 times by icing, partial freezing and freezing, respectively, as much as that of control product.

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Characterization of α-D-manosidase activity from Bacillus safensis MA-01 (Bacillus safensis MA-01 유래 알파-만노사이데이즈의 효소학적 특성)

  • Lee, Bo Mi;Kim, Joo Won;Park, Jae Kweon
    • Journal of Marine Bioscience and Biotechnology
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    • v.7 no.1
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    • pp.11-18
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    • 2015
  • An extracellular alkaline ${\alpha}$-D-mannosidase produced by a strain named as MA-01 was produced and its preliminary enzyme activity was characterized. Upon determining the 16S rDNA sequence and its homology search, the strain was identified to be one of species of the Bacillus safensis. Localization of enzyme was elucidated that ${\alpha}$-D-mannosidase can be found in culture medium as an extracellular enzyme. In addition, partial enzyme activity of 63% compared with the extracellular enzyme activity was observed in membrane protein. The optimal pH and temperature of the ${\alpha}$-D-mannosidase were pH 7.5 and $37^{\circ}C$, respectively. The $K_m$ and $V_{max}$ values of the ${\alpha}$-D-mannosidase in crude enzyme toward p-nitrophenyl-${\alpha}$-D-mannopyranoside were determined to be $455.6{\mu}M$ and $10.8{\mu}mole/min/mg$ of protein, respectively. To the best of our knowledge, this is the first report described the alkaline ${\alpha}$-D-mannosidase from the family of B. safensis.

Cloning and Sequencing of a Gene Involved in the Biosynthesis of Exopolysaccharide in Zoogloea Ramigera 115SLR (Zoogloea Ramigera 115SLR로부터 다당류 생합성에 관여하는 유전자의 분리 및 염기서열 결정)

  • Sam-Pin Lee;Min Yoo
    • Biomedical Science Letters
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    • v.6 no.1
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    • pp.1-9
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    • 2000
  • To identity the genes responsible for the biosynthesis of exopolysaccharide, recombinant plasmids pUEX10 and pLEX10 were constructed from plasmid pLEX3 which was isolated from the recombinant cosmid library of Zoogloea ramigera 115. The complete nucleotide sequence of the 1.7 kb genomic DNA insert in plasmid pUEX10 was determined. Its analysis identified two open reading frames (ORF3 & ORF4) which could encode two proteins. The amino acid sequence derived from ORF3 showed the homology with gumC protein in Xanthomonas campestris as well as exoP protein in Rhizobium melizoti. The partial amino acid sequence of ORF4 showed the homology with polysaccharide export protein in Thermotoga maritima. Z. ramigera 115SLR and Z. ramigera 115SLR/pLEX10 showed the similar pattern for EPS production. Yield of exopolysaccharides produced by Z. ramigera 115SLR and Z. ramigera 115SLR/pLEX10 was 0.26% (w/v) and 0.16% (w/v), respectively.

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Partial replacement of soybean meal by white lupine seeds in the diet of dairy cows

  • Joch, Miroslav;Kudrna, Vaclav
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.6
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    • pp.957-964
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    • 2020
  • Objective: An experiment was conducted to determine the effect of partial replacement of soybean meal (SBM) by white lupine seeds (WLS) on milk yield and quality, feed efficiency and rumen fermentation of high-yielding dairy cows. Methods: Thirty multiparous cows of two breeds (20 Holstein and 10 Czech Pied cows) in early mid-lactation received three diets (treatments) in a 3×3 Latin square design with a 28-d period. The dietary treatments were as follows: CON (control total mixed ration with SBM, no WLS), WLS30 (30% of the SBM was replaced, on a dry matter basis, by WLS), and WLS50 (50% of the SBM was replaced by WLS). Results: Feed intake by the cows was not affected (p = 0.331) by the diets. Milk production decreased with increasing proportions of WLS in the diet. Cows fed WLS50 yielded approximately 1 kg/d (p<0.001) less milk than cows fed the CON diet. The proportions of milk fat (p = 0.640), protein (p = 0.507), and lactose (p = 0.709) were not altered by the diet. For milk fat, feeding with WLS50 reduced the proportion of total saturated fatty acids (p<0.001) and increased the proportion of total monounsaturated fatty acids (p<0.001), mainly through oleic acid (p<0.001). No differences were found in feed efficiency, body weight, and blood plasma metabolites between groups. Rumen ammonia-N levels tended (p = 0.087) to increase with increasing proportions of WLS in the diet, whereas no effect of diet on rumen pH was found (p = 0.558). Conclusion: We did not identify the safe range within which raw WLS can efficiently replace SBM in the diet of high-producing dairy cows. In contrast, even partial replacement of SBM by WLS favorably changed the milk fatty acid profile.

Partial replacement of soybean meal with different protein sources in piglet feed during the nursery phase

  • Genova, Jansller Luiz;Carvalho, Paulo Levi de Oliveira;Oliveira, Newton Tavares Escocard de;Oliveira, Aparecida da Costa;Gois, Franz Dias;Castro, Davi Elias de Sa e;Souza, Fabio Nicory Costa;Trautenmuller, Heloise;Santos, Liliana Bury de Azevedo dos;Leal, Isabela Ferreira
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.11
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    • pp.1725-1733
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    • 2019
  • Objective: Evaluate the partial replacement of soybean meal with different protein sources in piglet feed during the nursery phase in terms of digestibility of feed, nitrogen balance, growth performance and blood parameters. Methods: Experiment I involved 24 crossbred entire male pigs with an initial body weight (BW) of $18.28{\pm}0.7kg$ and used a randomized complete block design consisting of 3 treatments (fish meal, FM; soybean protein concentrate, SPC; and soybean meal, SBM) and 8 replicates, with 1 pig per experimental unit. Experiment II involved 1,843 crossbred male and female pigs with an initial BW of $6.79{\pm}0.90kg$ and was based on a completely randomized design with a $2{\times}3$ factorial arrangement (2 sexes and 3 protein sources) and 13 replicates. Results: The results of Exp. I indicate effects (p<0.05) of dietary protein sources on digestible protein (FM, 17.84%; SPC, 16.72%, and SBM, 18.13%) and on total nitrogen excretion (TNE, $g/kg\;BW^{0.75}/d$) in which pigs fed with SBM-based feed had TNE values that were 5.36% and 3.72% greater than SPC and FM, respectively. In the Exp. II, there was difference (p<0.01) between sexes in the pre-starter I and starter phases, and total period in average daily feed intake (ADFI), which were greater in females, and between the protein sources, ADFI, final weight and daily weight gain. For urea in the pre-starter II and starter phases and glucose in the pre-starter II phase, there was a difference (p<0.05) between protein sources and between sexes, in starter phase in urea concentrations (females: 57.11 mg/dL and males: 50.60 mg/dL). Conclusion: The use of SBM as only protein source influences larger TNE ($g/kg\;BW^{0.75}/d$), reduces the growth performance of piglets and increases plasma urea concentrations in prestarter II phase.

Synergistic efficacy of LBH and αB-crystallin through inhibiting transcriptional activities of p53 and p21

  • Deng, Yun;Li, Yongqing;Fan, Xiongwei;Yuan, Wuzhou;Xie, Huaping;Mo, Xiaoyang;Yan, Yan;Zhou, Junmei;Wang, Yuequn;Ye, Xianli;Wan, Yongqi;Wu, Xiushan
    • BMB Reports
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    • v.43 no.6
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    • pp.432-437
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    • 2010
  • LBH is a transcription factor as a candidate gene for CHD associated with partial trisomy 2p syndrome. To identify potential LBH-interacting partners, a yeast two-hybrid screen using LBH as a bait was performed with a human heart cDNA library. One of the clones identified encodes ${\alpha}B$-crystallin. Co-immunoprecipitation and GST pull-down assays showed that LBH interacts with ${\alpha}B$-crystallin, which is further confirmed by mammalian two-hybrid assays. Co-localization analysis showed that in COS-7 cells, ${\alpha}B$-crystallin that is cytoplasmic alone, accumulates partialy in the nucleus when co-transfected with LBH. Transient transfection assays indicated that overexpression of LBH or ${\alpha}B$-crystallin reduced the transcriptional activities of p53 and p21, respectively, Overexpression of both ${\alpha}B$-crystallin and LBH together resulted in a stronger repression of the transcriptional activities of p21 and p53. These results showed that the interaction of LBH and ${\alpha}B$-crystallin may inhibit synergistically the transcriptional regulation of p53 and p21.