• Title/Summary/Keyword: Amino acids and Proteins Analysis

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The Juhwangjam (JH) 3rd day of fifth instar silkworm exhibits antioxidative properties and prevents high-fat diet-induced hypercholesterolemia

  • Ji Hae, Lee;Yeon-Ji, Kim;Kyungho, Kim;Hyun Bok, Kim;HaeYong, Kweon
    • International Journal of Industrial Entomology and Biomaterials
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    • v.45 no.2
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    • pp.126-130
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    • 2022
  • The silkworm is a food material that can simultaneously ingest phytochemicals from mulberry leaves, proteins with essential amino acids, and fatty acids. They are known to have hypoglycemic properties; however, further functional investigation is needed. In this study, four varieties of 3rd day of fifth instar silkworm with different cocoon colors, namely Baegokjam (BG), Goldensilk (GS), Yeonnokjam (YN), and Juhwangjam (JH), were compared in terms of antioxidant properties and cholesterol-lowering effect. JH, which had the highest polyphenol content (+38% vs. GS, p<0.05) showed high antioxidant efficacy. Treatment with JH also resulted in the lowest cholesterol biosynthesis enzyme activity (28% vs. control, p<0.05). In the animal study, high-fat diet (HFD)-fed mice that were orally administered JH extract for 12 weeks showed lower body weight gain (-10.4% vs. HFD, p<0.05) and serum total cholesterol levels (-12.7% vs. HFD, p<0.05). Comparing the varieties, JH had the highest effect. In future studies, analysis of the active ingredients according to their variety should be done.

Cloning and characterization of Giardia intestinalis cyclophilin

  • Yu, Hak-Sun;Kong, Hyun-Hee;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.40 no.3
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    • pp.131-138
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    • 2002
  • The cyclophilins (Cyps) are family members of proteins that exhibit peptidylprolyl cis-trans isomerase (PPIase, EC 5.2.1.8) activity and bind the immunosuppressive agent cyclosprin A (CsA) in varying degrees. During the process of random sequencing of a cDNA library made from Giardia intestinalis WB strain, the cyclophilin gene (gicypl) was isolated. An open reading frame of gicyp1 gene was 576 nucleotides, which corresponded to a translation product of 176 amino acids (Gicypl). The identity with other Cyps was about 58-71%. The 13 residues that constituted the CsA binding site of human cyclophilin were also detected in the amino acid sequence of Gicypl, including tryptophan residue essential for the drug binding. The single copy of the gicypl gene was detected in the G. intestinalis chromosome by southern hybridization analysis. Recombinant Gicyp 1 protein clearly accelerated the rate of cis ${\rightarrow}$ trans isomerization of the peptide substrate and the catalysis was completely inhibited by the addition of $0.5{\;}{\mu}M$ CsA.

Mutagenicity of Pyrolytic Products of Korean Animal Ptotein Foods by Salmonella/Mammalian-Microsome Test 1. Mutagenicity of Korean-Style Beef "Bulgogi" (Salmonella/Mammalian Microsome 실험법에 의한 육류식품 열분해산물의 돌연변이 유발능에 관한 연구 1. 한국 "불고기"의 돌연변이능)

  • 이미숙;모수미;박상철
    • Environmental Mutagens and Carcinogens
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    • v.7 no.2
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    • pp.85-102
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    • 1987
  • In spite of high incidences in gastrointestinal cancers in Korea, there have been few studies so far, which tried to detect or isolate the mutagenic, namely the carcinogenic substances in the Korean taste traditional foods. Among the many plausible dietary sources of carcinogens, the pyrolytic products of proteins or amino acids would be the major ones. And it is more noteworthy that the food pattern analysis showed the increasing consumption of protein foods by Korean as pyrolytic products, such as Bulgogi and Kalbi, etc.

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Protein Composition of Domestic and Glyphosate-Tolerant Soybean (국내산 및 제초제 내성 콩의 단백질 조성 분석)

  • Wei, Chun-Hua;Sok, Dai-Eun;Yang, Yun-Hyoung;Oh, Sang-Hee;Kim, Hyoung-Chin;Yoon, Won-Kee;Kim, Hwan-Mook;Kim, Mee-Ree
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.4
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    • pp.470-475
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    • 2006
  • In order to elucidate the differences of protein profiles among soybean cultivars, the protein composition of three conventional domestic soybean cultivars and two imported ones including glyphosate-tolerant HS2906 was analyzed by total nitrogen measurement, amino acid analysis and PAGE/densitometry. There were no statistically significant differences in the levels of any amino acid, including aromatic amino acids, between glyphosale-tolerant soybean and the conventional soybean WS82. In the extraction of protein, the SDS/buffer system was more efficient than the defatting/water system. The SDS-PAGE/densitometry analysis showed that there was a similar profile of proteins among cultivars, although the amount of total protein ranged from 380.2 mg/g to 423.9 mg/g. In addition, there was no discernable difference of protein profile between glyphosate- tolerant soybean (total protein amount, 380.2 mg/g) and the conventional soybean WS82 (390.2 mg/g), although the amount of ${\beta}$-conglycinin (55 kDa) was lower in glyphosate-tolerant soybean. Meanwhile, the amount of 25 kDa protein was greater in domestic soybean cultivars than imported ones. Thus, normal PAGE/ densitometry method would be useful to analyze the difference in protein profiles of soybean proteins, and furthermore Evaluate the protein profile of proteins between GMO and conventional soybean.

The Distribution of Selenium in Proteins of Saccaromyces Cerevisia and Analysis of Selenomethionine in Specific Protein (효모단백질내의 셀레늄 분포 및특정단백질내의 셀레노메티오닌 분석)

  • Shim Heeyoung Shim;Sangwook Ahn;Yonghyun Ahn
    • Journal of the Korean Chemical Society
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    • v.47 no.4
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    • pp.363-369
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    • 2003
  • Selenized yeast (Se yeast) containing $0.1{\%}$(w/w) of selenium was obtained when the yeast was incubated at a selenium concentration of 1$1.14{\times}10_-3 M$ in rich medium. After washing several times, the inorganic selenium on the cell wall was confirmed with MBRT. There was no indication of inorganic selenium on the cell wall when the blue color in MBRT was stayed for 15 minutes. The selenized yeast was sonicated, then the selenium contained protein was obtained after salting out by ammonium sulfate at the concentration $80{\%}$ saturation. The seven protein bands were seperated by SDS-PAGE and the selenium concentration in protein was measured by ICP-AES. Analytical data showed that the large expressed protein band contained a relatively large amount of selenium. The proteins of the 47kDa was contained the concentrations of 69.5 ${\mu}$ Se/g of most many content. The protein (47 kDa) was seperated from PVDF membrane by tank-electroblotting. The isolated protein was hydrolyzed under acid condition and reacted with PITC. The derivatives of amino acids were analyzed by HPLC and compared with the data obtained from regular yeast. The resulting selenium-yeast was analyzed with the selenomethionine concentration of $2{\%}$ comparaed with general amino acids. The goal of this study is to analyze the selenium concentration in protein bands and measure the degree of biotransformation of selenomethionine in a specific protein.

Three transcripts of EDS1-like genes respond differently to Vitis flexuosa infection

  • Islam, Md. Zaherul;Yun, Hae Keun
    • Journal of Plant Biotechnology
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    • v.44 no.2
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    • pp.125-134
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    • 2017
  • Enhanced disease susceptibility1 (EDS1) is a regulator of basal defense responses required for resistance mediated by TIR-NBS-LRR containing R proteins. We identified three transcripts of EDS1-like genes encompassing diverse/separate expression patterns, based on the transcriptome analysis by Next Generation Sequencing (NGS) of V. flexuosa inoculated with Elsinoe ampelina. These genes were designated VfEDL1 (Vitis flexuosa Enhanced Disease Susceptibility1-like1), VfEDL2 and VfEDL3, and contained 2464, 1719 and 1599 bp, with 1791, 1227 and 1599 bp open reading frames (ORFs), encoding proteins of 596, 408 and 532 amino acids, respectively. The predicted amino acid sequences of all three genes showed the L-family lipase-like domain (class 3 lipase domain), and exhibited a potential lipase catalytic triad, aspartic acid, histidine and serine in the conserved G-X-S-X-G. All three VfEDL genes were upregulated at 1 hpi against the bacterial and fungal pathogens Rizhobiumvitis and E. ampelina, respectively, except VfEDL1, which was downregulated against E. ampelina at all time points. Against E. ampelina, VfEDL2 and VfEDL3 showed downregulated expression at later time points. When evaluated against R. vitis, VfEDL1 showed downregulated expression at all time points after 1 hpi, while VfEDL3 showed upregulation up to 24 hpi. Based on the expression response, all three genes may be involved in plant resistant responses against R. vitis, and VfEDL2 and VfEDL3 show additional resistant responses against E. ampelina infection.

Identification and Cloning of jipA Encoding a Polypeptide That Interacts with a Homolog of Yeast Rad6, UVSJ in Aspergillus nidulans

  • Cho, Jae-Han;Yun, Seok-Soong;Jang, Young-Kug;Cha, Mee-Jeong;Kwon, Nak-Jung;Chae, Suhn-Kee
    • Journal of Microbiology
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    • v.41 no.1
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    • pp.46-51
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    • 2003
  • RAD6 in yeast mediates postreplication DNA repair and is responsible for DNA-damage induced mutations. RAD6 encodes ubiquitin-conjugating enzyme that is well conserved among eukaryotic organisms. However, the molecular targets and consequences of their ubiquitination by Rad6 have remained elusive. In Aspergillus nidulans, a RAD6 homolog has been isolated and shown to be an allele of uvs). We screened a CDNA library to isolate UVSJ-interacting proteins by the yeast two-hybrid system. JIPA was identified as an interactor of UVSJ. Their interaction was confirmed in vitro by a GST-pull down assay. JIPA was also able to interact with mutant UVSJ proteins, UVSJl and the active site cysteine mutant UVSJ-C88A. The N- and the C-terminal regions of UVSJ required for the interaction with UVSH, a RAD18 homolog of yeast which physically interacts with Rad6, were not necessary for the JIPA and UVSJ interactions. About 1.4 kb jipA transcript was detected in Northern analysis and its amount was not significantly increased in response to DNA-damaging agents. A genomic DNA clone of the jipA gene was isolated from a chromosome I specific genomic library by PCR-sib selection. Sequence determination of genomic and cDNA of jipA revealed an ORF of 893 bp interrupted by 2 introns, encoding a putative polypeptide of 262 amino acids. JIPA has 33% amino acid sequence identity to TIP41 of Saccharomyces cerevisiae which negatively regulates the TOR signaling pathway.

Development of animal protein(feed for fry) utilizing the rumen ciliates (제1위 섬모충(rumen ciliates)을 이용한 동물성 단백질(치어용 사료) 개발)

  • Jee, Cha-ho;Hyun, Gong-yool
    • Korean Journal of Veterinary Research
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    • v.35 no.2
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    • pp.327-336
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    • 1995
  • This study was carried out to develop the animal protein(feed for fry) that was isolated, purified and lyophilized the rumen ciliates from the healthy rumen contents which have $10^5-10^6/g$ ciliates and were discarded in abattoirs. The rumen ciliates are non-pathogenic, anaerobic and the weight of this protozoa is 2% of rumen content. The rumen protozoan and bacterial proteins both have a biological value for rats of 80-81, which is higher than the 72 of brewer's yeasts. Furthermore, the true digestibility and net protein utility of the protozoan protein are 91 and 73, much higher than those of bacterial(74 and 60) or yeast(84 and 60) proteins. The amino acids of rumen protozoa is nutritionally superior than the others. The size of rumen ciliates is $30-200{\times}20-110{\mu}m$ and so we had isolated and purified the rumen ciliates from the rumen contents by the physical methods. The purified rumen protozoa was lyophilized with freezing dryer. The results of this experiment were as follows : 1. Population dynamics of protozoan ciliates in slaughtered rumens; % of samples which small ciliates were predominated was 82.5%(52/63) and that of large ciliates was 17.5%(11/63). 1) predominant species of small ciliates were Entodinium ovinum and E nanellum. 2) predominant species of large ciliates were Epidinium ecaudatum and Diploplastron affine. 2. The lyophilized rumen ciliates which were isolated and purified from 1 kg of rumen content at the pH 6.2-6.8 was about 7.0 gram. 3. The nutrient analysis of lyophilized rqmen ciliates(LRC) was as follows: 1) Proximate analysis of the LRC and the composition of fry feed; moisture 8.05%(below 10.0), protein 35.37%(45), fat 5.39%(4.5), fiber 1.23%(below 2.5), ash 2.25%(below 15.0), Ca 0.26%(below 2.0), P 0.14%(below 1.1), energy 4,608.11(fish meal 5000 cal/g) 2) Amino acids (% in crude protein) of the LRC and the rotifer(Brachionus plicatilis); Arg 5.19%(4.50), His 2.50%(1.55), Ile 5.29%(3.45), Leu 8.11%(5.85), Lys 10.34%(6.15), Met 2.25% (0.85), Phe 5.66%(3.80), Thr 5.14% (3.45), Val 4.18%(3.90), Ala 4.13%(3.35), Asp 13.26%(8.25), Glu 16.62%(9.20), Gly 4.23%(3.10), Pro 3.25%(5.05), Ser 4.85%(3.85), Tyr 5.04%(3.05) 3) Fatty acids(% in fat) of the LRC and the rotifer(biological feed ; Brachionus plicatilis); myristic acid(C14:0) 3.27%(0.3), myristoleic acid(C14:1) 0.83%(-), palmitic acid(C16:0) 39.11% (23.5), palmitoleic acid(C16:1) 2.81%(2.0), stearic acid(C18:0) 9.36%(5.6), oleic acid(C18:1) 25.54%(3.5), linoleic acid(C18:2) 15.05%(32.9), linolenic acid(C18:3) 1.74%(9.8). Judging from the above investigated results, the analytical data of proximate analysis, amino acids, fatty acids of the purified and lyophilized rumen protozoa are reasonable for the feed of freshwater fishes(fry and fingerling). But it was disappointed of our expectation that the crude protein of lyophilized rumen ciliates contains low percentage, it was thought that because of the small ciliates(starch digester) in beef cattle rumens which were administered the concentrated feed, is much difficult to isolate and purify than the large ciliates(fiber digester).

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Identification of Retinol-binding Protein Produced by Caprine Endometrium during Periattachment Period of Early Pregnancy

  • Liu, K.H.;Huang, J.C.;Lin, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.12
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    • pp.1708-1713
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    • 2002
  • Endometrial explants obtained from does between days 13 and 21 of pregnancy were cultured in a modified minimum essential medium in the presence of [$^35S$]methionine and [$^3H$]-leucine. Proteins synthesized and secreted into medium were analyzed by fluorography of two-dimensional polyacrylamide gel electrophoresis and fluorography. No marked qualitative changes in patterns of protein production by caprine endometrium between days 13-21 of pregnancy. At least 11 proteins showed consistently a clear spot or a grouping of spots with characteristic location on two-dimensional gels. A major low molecular weight protein consisted of two major isoforms (pI 5.3-6.0) of similar molecular mass (21 kDa). Limited N-terminal sequence analysis of these two isoforms showed that the protein had complete homology with bovine placental and plasma retinol-binding protein (RBP) over the first 20 amino acids. Through use of the antiserum raised against bovine placental RBP, immunoreactive RBP was detected in cultures conditioned by uterine explants prepared at days 13, 15 and 21 of pregnancy. In the present study, proteins synthesized and secreted by caprine endometrium during periattachment period of early pregnancy were characterized. The pregnant endometrium secreted a number of neutral-to-acidic proteins which constituted, in part, the histotroph. A vitamin A-transport protein, RBP, was identified in cultures conditioned by endometrium of days 13-21 of pregnancy. The uterine endometrium is the only source of retinol for embryonic tissues. The uterine RBP appears to transport retinol locally toward embryonic tissues. Secretion of RBP by caprine endometrium of days 13, 15 and 21 of pregnancy suggested that retinol played an important role in conceptus development during periattachment period of early pregnancy.

Identification and Characterization of Two New S-Adenosylmethionine-Dependent Methyltransferase Encoding Genes Suggested Their Involvement in Stipe Elongation of Flammulina velutipes

  • Huang, Qianhui;Mukhtar, Irum;Zhang, Yelin;Wei, Zhongyang;Han, Xing;Huang, Rongmei;Yan, Junjie;Xie, Baogui
    • Mycobiology
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    • v.47 no.4
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    • pp.441-448
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    • 2019
  • Two new SAM-dependent methyltransferase encoding genes (fvsmt1 and fvsmt2) were identified from the genome of Flammulina velutipes. In order to make a comprehensive characterization of both genes, we performed in silico analysis of both genes and used qRT-PCR to reveal their expression patterns during the development of F. velutipes. There are 4 and 6 exons with total length of 693 and 978 bp in fvsmt2 and fvsmt1, respectively. The deduced proteins, i.e., FVSMT1 and FVSMT2 contained 325 and 230 amino acids with molecular weight 36297 and 24894 Da, respectively. Both proteins contained a SAM-dependent catalytic domain with signature motifs (I, p-I, II, and III) defining the SAM fold. SAM-dependent catalytic domain is located either in the middle or at the N-terminal of FVSMT2 and FVSMT1, respectively. Alignment and phylogenic analysis showed that FVSMT1 is a homolog to a protein-arginine omega-N-methyltransferase, while FVSMT2 is of cinnamoyl CoA O-methyltransferase type and predicted subcellular locations of these proteins are mitochondria and cytoplasm, respectively. qRT-PCR showed that fvsmt1 and fvsmt2 expression was regulated in different developmental stages. The maximum expression levels of fvsmt1 and fvsmt2 were observed in stipe elongation, while no difference was found in mycelium and pileus. These results positively demonstrate that both the methyltransferase encoding genes are involved in the stipe elongation of F. velutipes.