• 제목/요약/키워드: Amino acid sequence

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Characteristics of Chitosanases from Aspergillus fumigatus KB-1

  • Eom, Tae-Kyoung;Lee, Kang-Man
    • Archives of Pharmacal Research
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    • 제26권12호
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    • pp.1036-1041
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    • 2003
  • Two chitosanases produced by Aspergillus fumigatus KB-1 were purified by ion exchange and size exclusion chromatographies. Molecular weights of chitosanases were 111.23 kDa (chitosanase I) and 23.38 kDa (chitosanase II). The N-terminal amino acid sequence of chitosanase II was determined as follows: YNLPNNLKQIYDKHKGKXSXVLAKGFTN. The optimum pH of the chitosanase I and II was 6.5 and 5.5, respectively. The optimum temperatures were $60^{\circ}C$ for chitosanase land $70^{\circ}C$ for chitosanase II. Hydrolysis products of two chitosanases were analyzed by HPLC and GPC. Chitosanase I hydrolyzed substrate to glucosamine. Chitosanase II produced chitooligosaccharides.

Cloning and Functional Verification of the Candida milleri HIS3 Gene Encoding Imidazoleglycerol Phosphate Dehydratase

  • Park, Eun-Hee;Kwun, Se-Young;Han, Seung-Ah;Lee, Jong-Sub;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • 제22권10호
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    • pp.1441-1445
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    • 2012
  • The entire nucleotide sequence of the HIS3 gene encoding imidazoleglycerol phosphate dehydratase (IGPD) in yeast Candida milleri CBS 8195 was determined. Sequence analysis revealed an open-reading frame of C. milleri HIS3 that spans 678 bp, encodes 226 amino acids, and shares 80.5% amino acid identity to Torulaspora delbrueckii IGPD, followed by that to Saccharomyces cerevisiae (79.6%). The cloned HIS3 gene complemented a his3 mutation in S. cerevisiae, suggesting that it encodes a functional IGPD in C. milleri CBS 8195. A new auxotrophic marker is now available for acid-tolerant yeast C. milleri.

Cloning of Cellobiohydrolase Gene (cbhI) in Radiation Induced Mutant of Pleurotus florida

  • Lee, Young-Keun;Sathesh-Prabu, Chandran;Kim, Min Kyung
    • 방사선산업학회지
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    • 제8권2호
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    • pp.65-69
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    • 2014
  • The cellobiohydrolase gene (cbhI), a key component of a cellulolytic system, of a mutant PfCM4 (Pleurotus florida), developed through gamma ray radiation mutagenesis, was isolated and cloned. The deduced amino acid sequence was closely related to the glycoside hydrolase family 7 (GH7). The molecular weight of the deduced amino acid sequence of cbhI gene was found to be 22.4 kDa. Though the percent identity was found to be much less (35.61%) between the wild type and mutant, the cellulolytic activity of PfCM4 was 17.24% higher than that of the wild type. This shows that the catalytic domain of the cbhI gene was conserved in the mutant PfCM4.

Bacillus subtilis subsp. krictiensis가 생산하는 항진균 물질 KRF-001의 구조 결정 (Structure Determination of Antifungal KRF-001 Produced by Bacillus subtilis subsp. krictiensis)

  • 김성기;이남경;정태숙;김영국;최진자;복성해
    • 한국미생물·생명공학회지
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    • 제19권6호
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    • pp.598-603
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    • 1991
  • 신규 미생물 Bacillus subtilis subsp. krictiensis의 배양액으로부터 식물 및 인체 병원균에 폭넓은 항진균 활성을 나타내는 DRF-001으로 총칭되는 6개의 cyclic peptide의 복합체(A에서 F)를 분리하였다. 이 6개 peptides의 분자량은 FAB-MS 측정결과, A가 1042, B와 C가 1056, D와 E가 1070 그리고 F가 1084였다. 이 복합체의 구조를 각종 자기분석을 통해 해석한 결과, 구조 중에 1몰씩의 glutamine, proline, tyrosine, serine 및 unusal $\beta$-amino acid와 3몰의 asparagine을 공통적으로 갖고 있으나, $\beta$-amino acid의 탄소수와 말단 methyl 구조에 차이점이 있음을 알 수있었다. 구성 amino acid들의 서열과 입체구조 결정에 의하여 KRF-001의 잠정구조를 결정하였다.

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Genotypic Diversity of the Complete Open-Reading Frame 7 Sequences of Porcine Reproductive and Respiratory Syndrome Viruses in Korea and Coexistence of Two Genotypes

  • Chu, Jia-Qi;Kim, Myung-Cheol;Park, Chang-Sik;You, Myung-Jo;Jun, Moo-Hyung
    • 한국임상수의학회지
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    • 제25권3호
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    • pp.139-145
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    • 2008
  • To investigate the genotypic diversity of the porcine reproductive and respiratory syndrome viruses (PRRSV) in Korea, we examined 92 clinical samples from three provinces by RT-PCR and a nested PCR, and the complete open-reading frame 7 (ORF 7) sequences of 15 samples selected from 72 PCR-positive specimens were analyzed. When we compared nucleotide (amino acid) sequences of 80 isolates from Korea and overseas countries, the sequences of 7 samples belonged to North American (NA)-genotype, and those of 8 samples, to European (EU)-genotype. The nucleotide (amino acid) identities between two genotypes were 63.7% (59.8%) to 65.1% (63.1%). When compared with NA prototype VR-2332, the 7 strains of NA-genotype shared 89.8% (93.6%) to 91.2% (96.0%) identity of nucleotide (amino acid) sequence. The 8 strains of EU-type shared 93.6% (92.3%) to 94.3% (93.8%) identity of nucleotide (amino acid) sequence as compared to EU prototype Lelystad. In phylogenetic tree analysis by neighbor-joining method, all of the 8 EU-type strains were clustered into group 4 distinct from ED-prototype Lelystad (group 1). In NA-genotype, 24 domestic isolates reported previously and the 7 strains of NA-type determined in this study were clustered into group 1, while US prototype VR 2332 was classified into different group (group 2). These results suggest that emergence of EU-genotype and the dual-infection of NA- and EU-genotypes may be prevalent in the pig farms in Korea. The high degree of genetic diversity of field PRRSVs should be taken into consideration for control and preventive measures.

Endophytic Bacillus sp. CY22 from a Balloon Flower (Platycodon grandiflorum) Produces Surfactin Isoforms

  • Cho, Soo-Jeong;Hong, Su-Young;Kim, Jin-Young;Park, Sang-Ryeol;Kim, Min-Keun;Lim, Woo-Jin;Shin, Eun-Chule;Kim, Eun-Ju;Cho, Yong-Un;Yun, Han-Dae
    • Journal of Microbiology and Biotechnology
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    • 제13권6호
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    • pp.859-865
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    • 2003
  • Surfactin is a mixture of cyclic lipopeptides built from variants of a heptapeptide and a ${\beta}-hydroxy$ fatty acid produced by several strains of Bacillus sp. Surfactin isoforms produced by endophytic Bacillus sp. CY22 from a balloon flower were isolated and characterized. It was found that the purified surfactin had three isoforms with protonated masses of m/z 1,008, 1,022, and 1,036, and different structures in combination with Na, K, Ca ions using MALDI-TOF MS, ESI-MS/MS, and ICP MS, respectively. In the MS/MS analysis, the isolated surfactin had the identical amino acid sequence (LLVDLL) and hydroxy fatty acids (with 13 to 15 carbons in length), even though isolated from different Bacillus strains. The sfp22 gene, required for producing the surfactin, consisted of an open reading frame (ORF) of 675 bp encoding 224 amino acid residues with a signal peptide of 20 amino acids. The predicted amino acid sequence of sfp22 was very similar to that of Ipa-8.

Structural Characteristics of the Putative Protein Encoded by Arabidopsis AtMTN3 Gene

  • Cheong, Jong-Joo;Kwon, Hawk-Bin;Kim, Minkyun
    • Journal of Applied Biological Chemistry
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    • 제44권3호
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    • pp.125-130
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    • 2001
  • A putative protein encoded by Arabidopsis AtMTN3 gene, a homologue of Medicago truncatula MTN3, consists of 285 amino acid residues, and has a predicted molecular mass of 31.5 kDa and a calculated pI of 9.1. Primary amino acid sequence analyses have revealed that the protein contains seven putative transmembrane regions with N-terminus oriented to the outside of the membrane. The AtMTN3 protein shows overall 16.4% of amino acid identity with the rat GALR3 protein, known to be a G-protein-coupled receptor. The gene is present as a single copy in the Arabidopsis genome, and expressed in aerial parts but not in roots of Arabidopsis. Therefore, AtMTN3 appears not to be specifically involved in Rhizobium-induced nodule development, as was predicted for the MTN3 gene. These proteins possibly mediate signal transmission through G-protein-coupled pathways during general interactions between plants and symbiotic or pathogenic microbes.

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Identification of Amino Acid Residues in the Carboxyl Terminus Required for Malonate-Responsive Transcriptional Regulation of MatR in Rhizobium leguminosarum bv. trifolii

  • Lee, Hwan-Young;Kim, Yu-Sam
    • BMB Reports
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    • 제34권4호
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    • pp.305-309
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    • 2001
  • MatR in Rhizobium trifolii is a malonate-responsive transcription factor that regulates the expression of genes, matABC, enabling decarboxylation of malonyl-CoA into acetyl-CoA, synthesis of malonyl-CoA from malonate and CoA, and malonate transport. According to an analysis of the amino acid sequence homology, MatR belongs to the GntR family The proteins of this family have two-domain folds, the N-terminal helix-turn-helix DNA-binding domain and the C-terminal ligand-binding domain. In order to End the malonate binding site and amino acid residues that interact with RNA polymerase, a site-directed mutagenesis was performed. Analysis of the mutant MatR suggests that Arg-160 might be involved in malonate binding, whereas Arg-102 and Arg-174 are critical for the repression activity by interacting with RNA polymerase.

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A Statistical Thermodynamic Study on the Conformational Transition of Oligopeptide Multimer

  • 김용구;박형석
    • Bulletin of the Korean Chemical Society
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    • 제17권2호
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    • pp.131-138
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    • 1996
  • The conformational transition of oligopeptide multimer,-(HPPHPPP)n-, is studied (H:hydrophobic amino acid, P:hydrophilic amino acid). The helix/coil transitions are detected in the multimer. The transition depends on the number of amino acid in the sequence, the concentration of the oligopeptide, and temperature which affects helix stability constant (${\xi}$) and hydrophobic interaction parameter (wj). In the thermodynamic equilibrium system jA${\rightarrow}$Aj (where A stands for oligopeptide monomer), Skolnick et al., explained helix/coil transition of dimer by the matrix method using Zimm-Bragg parameters ${\xi}$ and $\sigma$ (helix initiation constant). But the matrix method do not fully explain dangling H-bond effects which are important in oligopeptide systems. In this study we propose a general theory of conformational transitions of oligopeptides in which dimer, trimer, or higher multimer coexists. The partition of trimer is derived by using zipper model which account for dangling H-bond effects. The transitions of multimers which have cross-linked S-S bonds or long chains do not occur, because they keep always helical structures. The transitions due to the concentration of the oligopeptides are steeper than those due to the chain length or temperature.

Effects of Guanidination with Trypsin, Lys-C, or Glu-C Digestion on Mass Spectrometric Signal Intensity and Protein Sequence Coverage

  • Han, Hye-Sun;Nho, Seon-Ho;Lee, Ae-Ra;Kim, Jeong-Kwon
    • Bulletin of the Korean Chemical Society
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    • 제31권6호
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    • pp.1527-1534
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    • 2010
  • The conventional peptide modification process of guanidination, in which the amino groups of lysine residues are converted to guanidino groups using O-methylisourea to create more basic homoarginine residues, is often used to improve the signal intensity of lysine-containing peptides in matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Here, we used three different protease enzymes (trypsin, Lys-C, and Glu-C) to evaluate the effects of guanidination on the MS signals of two enzymatically digested proteins. Horse heart myoglobin and bovine serum albumin were guanidinated either before or after digestion with trypsin, Lys-C, or Glu-C. The resulting peptides were subjected to MALDI-MS, and signal intensities and sequence coverage were systematically evaluated for each digest. Guanidination prior to Glu-C digestion improved sequence coverage for both proteins. For myoglobin, guanidination before enzymatic digestion with trypsin or Lys-C also enhanced sequence coverage, but guanidination after enzymatic digestion enhanced sequence coverage only with Lys-C. For albumin, guanidination either before or after Glu-C digestion increased sequence coverage, whereas pre- or post-digestion guanidination decreased sequence coverage with trypsin and Lys-C. The amino acid composition of a protein appears to be the major factor determining whether guanidination will enhance its MALDI-MS sequence coverage.