• Title/Summary/Keyword: 구성아미노산

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Physicochemical Characteristics of Filefish and Cod Skin Collagen (말쥐치피 및 대구피 콜라겐의 물리화학적 특성)

  • Kim, Se-Kwon;Kang, Ok-Ju;Kwak, Dong-Chae
    • Applied Biological Chemistry
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    • v.36 no.3
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    • pp.163-171
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    • 1993
  • The collagens from filefish (Novoden modestus) and cod (Gadus macrocephalus Tilesius) skin were isolated and their physicochemical properties were investigated. Glutamic acid, hydroxyproline, valine and phenylalanine in the filefish skin collagen (FSC) were presented at higher levels than those of cod skin collagen (CSC), but the contents of glycine, proline and serine were contrary. The content of essential amino acids of FSC (265 residues/1000 residues) was higher than CSC (229 residues). The solubilities of both collagens were the lowest at pH 7.0, but precipitously increased at acid zone(below pH 5.0). FSC has lower viscosity than CSC. Furthermore, while the viscosities of both collagens were the lowest at pH 7.0, the viscosities of FSC and CSC were the highest at pH 4.0 and pH 2.0, respectively. The denaturation temperature of $FS(25^{\circ}C)$ was higher than $CSD\;(17^{\circ}C)$. The free hydrophobic residue contents of FSC and CSC tended to increase till $60^{\circ}C,\;and\;50^{\circ}C$ respectively, and to decrease thereafter. Hydration capacities of both collagens were the lowest at pH 7.0, and CSC had the superior hydration capacity to FSC. In addition, emulsifying and emulsifying stability of CSC was also superior to FSC.

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Analysis of a Region Required for the Functions of Fission Yeast Nucleoporin Nup184 and Its SUMO Modification (분열효모 핵공단백질인 Nup184의 기능에 필요한 부위 분석 및 SUMO 변성)

  • Chai, Ai-Ree;Jang, Soo-Yeon;Yoon, Jin-Ho
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.66-72
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    • 2012
  • The Nup188 protein is one of the largest evolutionally conserved nucleoprins (Nups) that compose the inner ring of nuclear pore complex (NPC). The Nup184 protein, fission yeast Schizosaccharomyces pombe ortholog of Nup188p, is required for normal growth and mRNA export in nutrient-rich medium (YES). Here, we identified a carboxyl region (482 to 1628) of Nup184 protein that was enough to complement the defects of both growth and mRNA export when the ${\Delta}nup184$ knock-out mutant was grown in YES medium. This region is also required for localization of GFP-Nup184 fusion to the nuclear periphery. In addition, we found that ORF of Nup184 (predicted 1564 amino-acid protein) registered in S. pombe GeneDB (hosted by Sanger Institute, UK) is 64 amino-acid residues shorter than that predicted by our sequence data. This carboxy-terminal region is necessary for the functions of Nup184p. We further demonstrated that Nup184 protein was conjugated with SUMO in vivo.

Complete Nucleotide Sequence Analysis and Structural Comparison of 3 members of Tomato Phenylalanine ammonia-lyase gene (토마토에서 분리한 3종류의 Phenylalanine ammonia-lyase gene에 대한 염기서열 및 특성비교)

  • 여윤수;예완해;이신우;배신철;류진창;장영덕
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.1
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    • pp.41-47
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    • 1999
  • Phenylalanine ammonia-lyase (PAL; EC 4, 3, 1, 5) genomic clones were isolated from tomato(Lycopersicon esculentum L.) genomic DNA libraries using tomato PAL5 cDNA sequences as probes. The nucleotide sequences of tPAL1, tPAL4 and tPAL5 were compared. tPAL5 contains an open reading frame encoding a polypeptide of 722 amino acids, interrupted by a 710 bp intron in the codon for the amino acid 139. tPAL1 encodes a polypeptide of 249 amino acids which is much shorter than tPAL5 gene due to a premature stop codon and does not contain an intron. tPAL4 encodes a polypeptide of 357 amino acids, interrupted by a 305 bp intron in the codon for the amino acid 138. Premature stop codons observed in tPAL1 and tPAL4 gene produce a short polypeptide rather than a normal polypeptide (722 aa). tPALl shows 87.2% homology with tPAL4 and 85.3% homology with tPAL5 gene whereas tPAL4 showes 91.4% homology with tPAL5 at nucleotide level. In general, phylogenetic analysis showed that genes isolated from tomato, potato, and sweet potato were belong to the same group and another dicot plants such as parsley, bean, soybean, pea and alfalfa formed another group. PAL genes isolated from rice and yeast showed very low homology with other PAL genes and formed the other group.

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Quality Characteristics of Eogeul-tang Added with Beef Gristle (도가니 연골 첨가에 따른 어글탕의 품질특성)

  • Moon, Min-Jung;Jin, So-Yeon;Kim, Myung-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.3
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    • pp.431-438
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    • 2014
  • Eogeul-tang is a traditional Korean soup dish made from dried pollack, minced beef, and tofu. This comparative study analyzed the quality characteristics of Eogeul-tang by varying the added amounts of cow gristle. The objective of this study was to analyze Eogeul-tang to promote its functionality and increased consumer preference. Collagen contents were $18.08{\pm}0.13g$ and $22.17{\pm}0.14g$ per 100 g of dried pollack skin and knee cartilage of cow, respectively. The overall collagen content was higher in the knee cartilage of cow. Different amounts of cow gristle (25%, 50%, 75%, and 100%) were added to traditionally cooked Eogeul-tang, and the general composition of Eogeul-tang was analyzed. Significant differences were exhibited in the amounts of water content, crude protein, crude fat, and crude ash depending on the added amounts of cow gristle. Moreover, the collagen content also significantly increased as the amount of gristle content increased. In particular, significant increases in the amounts of collagen components, including proline, glycine, and alanine were observed with increasing amount of gristle. When a sensory test was conducted on Japanese, overall preference values were highest for Eogeul-tang composed of 25% beef and 75% gristle compared to traditionally cooked Eogeul-tang. In conclusion, the study results promote the functionality of collagen as well as the increased quality of Eogeul-tang added with cow gristle manufactured by traditional cooking methods.

Isolation and Characterization of Eukaryotic Translation Initiation Factor 5A (eIF-5A) from Potato (감자로부터 Eukaryotic Translation Initiation Factor 5A (elF-5A) 유전자의 동정 및 발현 분석)

  • 인준교;신동호;최관삼;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.5
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    • pp.283-287
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    • 2001
  • Differential display based on PCR was employed to identify genes expressed during tuber-developing stage of potato (Solanum tuberosum L. cv. Irish Cobbler). An eukaryotic initiation factor 5A (eIF-5A) clone isolated from a cDNA library constructed with developing micro-tuber using a probe of PCR fragment. We isolated three positive clones and ore of them contained open reading frame. This clone revealed high sequence similarity to tomato eIF 5A cDNA. At the DNA level, there is 94.8% identity with the tomato eIF-5A4, whereas at the protein level there is a high identity with 97.5%. The potato eIF 5A clone is 716 bp in length and contains a single open reading frame from 57 to 539 bp, a 56 bp 5'-untranslated region and a 177 bp 3'-untranslated region. The deduced protein composed of 160 amino acid residues, with a predicted molecular mass of 17.4 kD and an estimated pl of 5.5. The sequence of 12 (STSKTGKHGHAK) amino acids among eIF-5A proteins is perfectly conserved from yeast to human. That sequence in potato eIF-5A protein is also conserved at position 46 to 57 amino acid. This region embeds the post-translational modification site of the lysine residue (at the seventh K) to hypusine that is crucial to eIF-5A activity. The northern blot analysis of eIF5A has shown abundant expression, mainly in flower organs (stamen, ovary, petal, sepal), fruit and stolen.

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Action mechanism of upstream open reading frame from S-adenosylmethionine decarboxylase gene as a in vivo translational inhibitor (S-Adenosylmethionine decarboxylase 유전자의 upstream open reading frame이 in vivo에서 translational inhibitor 로서의 작용 기작)

  • Choi, Yu-Jin;Park, Ky-Young
    • Journal of Plant Biotechnology
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    • v.38 no.1
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    • pp.87-93
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    • 2011
  • S-Adenosylmethionine decarboxylase (SAMDC; EC 4.1.4.50), a key enzyme for polyamines biosynthesis, was tightly regulated for homeostatic levels. Carnation SAMDC gene (CSDC9) has an small upstream open reading frame (uORF) of 54 amino acids in 5'-leader sequence. To explore the functional mechanism of uORFs in controlling translation, we used a GUS reporter gene driven with the 35S promoter and uORF region of SAMDC gene for making transgenic tobacco plants. In our experiment, there were a translational inhibition of its downstream GUS ORF by SAMDC uORF sequence or SAMDC uORF protein. Expecially, translational inhibition was most effective in point-mutated construct, in which the start codon was changed. Therefore, this results suggested the ribosomal stalling might be involved in this translational inhibitory process. The frame shift in amino acid sequence of SAMDC uORF with start codon and stop codon resulted in a moderate increasing in GUS activity, suggesting the native amino acid sequence was important for a function as a translational inhibitor. Also, we showed that the production of GUS protein was significantly inhibited in the presence of the small uORF using histochemical analysis of GUS expression in seedlings and tobacco flowers. Importantly, the small uORF sequence induced a real peptide of 5.7 kDa, which was provided the presence of SAMDC uORF peptide band using an in vitro transcription/translation system. The peptide product of uORF might interact with other components of translational machinery as well as polyamines, which was resulted from that polyamine treatment was inhibited GUS protein band in SDS-PAGE experiment.

Isolation and Identification of Chlorella sp. CMS-1 and the Chemical Composition of Its Hot Water Extract (클로렐라 균주의 분리 동정 및 열수 추출물의 이화학적 조성)

  • Cha, Jae-Young;Kim, Jung-Wook;Park, Bo-Kyung;Jin, Hyun-Jin;Kim, Sung-Young;Cho, Young-Su
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1723-1727
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    • 2008
  • Chlorella sp. CMS-1 strain was isolated from the outdoors cultivation pools in Culmansa Co., Ltd. This strain was found to be a rounded type of 3 ${\mu}m$. Phylogenetic analysis by the 18S rRNA sequencing using isolated strain is most similar to Chlorella sp. IFRPD 1018 gene at the level of nucleotide sequence identity at 99%. Accordingly, the isolated Chlorella strain was named as Chlorella sp. CMS-1 based on its morphological and phylogenetic properties. The concentrations of crude protein and fat were 59% and 0.01%, respectively. Major compositional amino acids (mg%) were glutamic acid 6.21, alanine 5.76, aspartic acid 5.44%, glycine 4.29%, and threonine 3.09% and major free amino acids (mg%) were ${\gamma}$-aminobutyric acid (GABA) 7.13%, L-alanine 1.44%, L-glutamic acid 0.90, L-leucine 0.26% and L-glycine 0.20%. The concentrations of major minerals were P 2.25%, K 2.25%, Na 1.09%, Mg 0.63%, and Ca 0.28%.

Expression of the Second Isoform of Gonadotropin-Releasing Hormone (Chicken GnRH-II Type) in the First Trimester Human Placenta (임신초기 사람의 태반조직에서 GnRH-II mRNA와 Peptide의 발현)

  • Cheon, Kang-Woo;Hong, Sung-Ran;Lee, Hyoung-Song;Kang, Inn-Soo
    • Development and Reproduction
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    • v.5 no.1
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    • pp.81-88
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    • 2001
  • Gonadotropin-releasing hormone (GnRH) has been known to play a role in the regulation of hCG secretion by human placenta. Recently, a gene encoding the second f개m of GnRH (GnRH-II) was identified in human. Herein, we demonstrate that GnRH-II is expressed in human placenta and assess GnRH-II expression by nested RT-PCR and immunohistochemistry in human placenta during the first trimester. We found that two altematively spliced transcripts of GnW-II mRNA were expressed in human placental tissues of first trimester and the shorter variant had a 21-bp deletion in GnRH-associated peptide (GAP). Immunoreactive GnRH-II was localized in both cytotrophoblastic and syncytiotrophoblastic cytoplasm. The immunostaining intensity was stronger in cytotrophoblast. Villous stromal cells also showed GnRH-II immunoreactiyiry. The results of our study report that the second isoform of GnRH (GnRH-II) is expressed in the first trimester human placenta and we suggest that GnRH-II may also play a regulatory role in maintenance of early pregnancy and hCG secretion in human placenta.

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Optimization of enzymatic hydrolysis of viscera waste proteins of black body fowl(Yeonsan Ogae) to produce peptides using a commercial protease and it's characters analysis (단백질 분해효소를 이용한 연산오계 내장 펩타이드 생산 최적화 및 특성분석)

  • Choi, So-Young;Kim, A-Yeon;Song, Yu-Rim;Ji, Joong-Gu;Yoo, Sun-Kyun
    • Journal of Digital Convergence
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    • v.14 no.1
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    • pp.253-262
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    • 2016
  • Yeonsan Ogae has been known as supporting health and high efficacy of treatment. In recent days, as the efficacy of functional peptides has known, the optimization of oligo peptides production and its characteristics from Ogae viscera has been performed. Response surface method was used to perform the optimizaion of enzyme hydrolysis. The range of processes was temperature (40, 50 and $60^{\circ}C$), pH(6.0, 7.0 and 8.0), and enzyme(1, 2 and 3%). The degree of hydrolysis, amono acids, molecular weight of products were analyzed. The optimum process of enzyme hydrolysis were determined as temperature $58^{\circ}C$, pH 7.5, and enzyme concetration 3%. At optimum conditions, the degree of hydrolysis after 2 h reaction was 75-80%. The total amino acids of amino acid and were 386.15 mg/100 g and 155.26 mg/100 g, respectively. The molecular weight of products by using Maldi-TOF was ranged from 300 to 1,000 Da.

The amino acid analysis of polyhedrin and DNA sequence of ployhedrin gene in nuclear polyhedrosis virus (Nuclear polyhedrosis virus의 polyhedrin 아미노산 및 polyhedrin gene 염기서열 분석)

  • Lee, Keun-Kwang
    • Journal of fish pathology
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    • v.8 no.1
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    • pp.37-46
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    • 1995
  • The amino acid analysis of polyhedrin protein and nucleotide sequence of polyhedrin gene in H. cunea nuclear polyhedrosis virus (HcNPV) genome have been studied. Polyhedrin had three polypeptide bands in SDS - polyactylamide gel electrophoresis. The major polypeptide had a molecular weight of 25 kd. The polyhedrin was composed of 17 different amino acids. HcNPV DNA was digested with EcoRI restriction enzyme and hybridized with ($\alpha^{32}P$) -labelled AcNPV polyhedrin gene cDNA. The polyhedrin gene was located on the fragment of EcoRI-H. The EcoRI - H fragment containing polyhedrin gene was cloned into the EcoRI site of pUC8 vector which was confirmed with southern blotting, and the recombinant plasmid containg polyhedrin gene was designated as hPE-H. The promoter region of polyhedrin genomic DNA was sequenced. The sequences identified as the TATA box was found at the 5' flanking region of the polyhedrin genomic DNA approximately -79 bp upstream from the transcriptional start site. But CAAT-like box was not shown near the TATA-like box in the polyhedrin gene. Four tandem repeats with the sequence 5' -CTAATAT-3' and 5'-TAAATAA-3' were found between -141 and -108 or -83 upstream and -52 bp downstream from the translation start site. About -141 bp region upstream from the translational start site was highly AT (78%) rich. The coding region for the polyhedrin starts and ends with ATG and TAA, respectively.

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