• Title/Summary/Keyword: Polypeptide

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Nucleotide Sequence and Characterization of ptsG Gene Encoding Glucose-specific Enzyme II of Phosphotransferase System from Brevibacterium flavum

  • Yoon, Ki-Hong
    • Journal of Applied Biological Chemistry
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    • v.48 no.4
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    • pp.218-221
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    • 2005
  • Nucleotide sequence of Brevibacterium flavum ptsG gene capable of complementing Escherichia coli ZSC113 mutations defective to glucose permease activity of phosphotransferase system was completely determined, and the gene product was compared with other glucose-specific enzyme II ($EII^{Glc}$). A ptsG gene of B. flavum consisted of open reading frame of 2,025 nucleotides putatively encoding polypeptide of 675 amino acid residues and TAA stop codon. Deduced amino acid sequence of B. flavum ($EII^{Glc}$) had high homology with ($EIIs^{Glc}$) of Corynebacterium glutamicum, C. efficiens, and B. lactofermentum. Arrangement of structural domains, IIBCA, of B. flanum ($EII^{Glc}$) protein was identical to that of EIIs belonging to glucose-phosphotransferase system.

Analysis of a Putative DNA Polymerase I gene in Brevibacterium ammoniagenes. (Brevibacterium ammoniagenes의 DNA Polymerase I 유사 유전자의 분석)

  • 오영필;윤기홍
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.105-110
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    • 2002
  • The sequence of 3,221 nucleotides immediately adjacent to rpsA gene encoding 30S ribosomal protein S1 of Brevibacterium ammoniagenes was determined. A putative open reading frame (ORF) of 2,670 nucleotides for a polypeptide of 889 amino acid residues and a TAG stop codon was found, which is located at a distance of 723 nucleotides upstream from rpsA gene with same translational direction. The deduced amino acid sequence of the ORF was found to be highly homologous to the DNA polymerase I of Streptomyces griseus (75.48%), Rhodococcus sp. ATCC 15963 (56.69%), Mycobacterium tuberculosis (55.46%) and Mycobacterium leprae (53.99%). It was suggested that the predicted product of the ORF is a DNA polymerase I with three functional domains. Two domains of 5 → 3 exonuclease and DNA polymerase are highly conserved with other DNA polymerase I, but 3 → 5 exonuclease domain is less conserved.

Cloning and Sequencing of the ddh Gene involved in the Novel Pathway of Lysine Biosynthesis from Brevibacterium lactofermentum

  • Kim, Ok-Mi;Kim, Hyun-Jeong;Kim, Dal-Sang;Park, Dong-Chul;Lee, Kap-Rang
    • Journal of Microbiology and Biotechnology
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    • v.5 no.5
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    • pp.250-256
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    • 1995
  • The ddh gene encoding meso-diaminopimelate (meso-DAP)-dehydrogenase (DDH) in Brevibacterium lactofermentum was isolated by complementation of the Escherichia coli dapD mutation. It was supposed from subcloning experiments and complementation tests that the evidence for DDH activity appeared in about 2.5 kb Xhol fragmented genome. The 2.5 kb Xhol fragment containing the ddh gene was sequenced, and an open reading frame of 960 bp encoding a polypeptide comprising 320 amino acids was found. Computer analysis indicated that the deduced amino acid of the B. lactofermentum ddh gene showed a high homology with that of the Corynebacterium glutamicum ddh gene.

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Identification of Multiple Active Forms in Cellulase-xylanase of Aspergillus sp. 8-17 by Active Staining

  • Shin, Pyung-Gyun;Ahn, Jun-Bae;Kim, Chang-Young;Jeong, Won-Hwa;Ryu, Jin-Chang
    • Journal of Microbiology and Biotechnology
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    • v.8 no.1
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    • pp.49-52
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    • 1998
  • A fungal strain able to produce filter paper activity (FPase) was isolated from soil by testing the ability to hydrolyze using filter paper. The isolated strain was identified as an Aspergilus sp. judging from its morphological and microscopical characteristics. The cellulase-xylanase system of Aspergillus sp. 8-17 was detected in situ after gel electrophoresis in the presence of SDS and showed that each protein pattern had a distinct polypeptide composition. ${\beta}$-1,4-Glucanase, cellobiohydrolase, and xylanase activity profiles differ from protein patterns. The Aspergillus sp. 8-17 hydrolytic enzymes responsible for the hydrolysis of ${\beta}$-glucan, MUC, and xylan have multiple active forms.

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Cloning and Expression of Inulin Fructotransferase Gene of Arthrobacter sp. A-6 in Escherichia coli and Bacillus subtilis

  • Kim, Hwa-Young;Kim, Chan-Wha;Choi, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.275-280
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    • 2000
  • The inulin fructotransferse (depolymerizing) (IFTase, EC 2.4.1.93) gene of Arthrobacter sp. A-6 was cloned and expressed in Escherichia coli and Bacillus subtilis. The IFTase gene consisted of an ORF of 1.311 nucleotides encoding a polypeptide of 436 amino acids containing a signal peptide of 31 amino acids in the N-terminus. The molecular mass of the IFTase based on the nucleotide sequence was calculated to be 46.116 Da. The recombinant E. coli $DH5{\alpha}$ cells expressing the Arthrobacter sp. A-6 IFTase gene produced most of the IFTase intracelularly. In contrast, the recombinant B. subtilis DB 104 carrying the IFTas gene on a B. subtilis-E. Coli expression vector secreted the IFTase into the culture fluid efficiently.

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Diffusional Behavior of Rodlike Poly(${\gamma}-benzyl$ L-glutamate) in Concentrated Solution as Studied by the Field-Gradient $^{1}H$ NMR Methods

  • Kuroki, Shigeki
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.345-345
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    • 2006
  • It is well known that poly(glutamate)s with long n-alkyl side chains form thermotropic liquid crystalline state by melting of the side-chain crystallites and also poly(glutamate)s such as poly(${\gamma}-benzyl\;L-glutamate$ )(PBLG), poly(${\gamma}-n-alkyl\;L-glutamate$), etc. in solvent form the isotropic, biphasic and liquid crystalline phases which contains cholesteric and columnar liquid crystalline forms depending on the polypeptide concentration. Although there is no diffusion study for PBLG in liquid crystalline state, because it is difficult to observed $^{1}H$ spectrum of PBLG in liquid crystalline state for its very short $^{1}H\;T_{2}$. In this study, the diffusional behavior of rodlike PBLG in concentrated solution is successfully elucidated as studied by the field-gradient $^{1}H$ NMR methods.

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Cloning and expression of glutathione S-transferase (GST) cDNA from Gossypium hirsutum L.

  • Kang, Won-Hee;Kim, Myong-Jo;Lim, Jung-Dae;Yun, Song-Joong;Chung, Ill-Min;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.4
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    • pp.294-297
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    • 2002
  • A gene coding for the GST of cotton (Gh-5) was cloned into Escherichia coli and experssed. The enzyme remained within the cytoplasm of E. coli. An 696 bp open reading frame was in the 988 base pair fragment of the recombinant plasmid pET-30b(+). The deduced protein sequence consists of 232 amino acids and has a molecular mass of 30235.58 Da. The cloned enzyme conjugated reduced glutathione and 1-chloro-2,4-dinitrobenzene (CDNB). Plant GST cDNA was expressed in microbe and produced polypeptide had function as an enzyme.

Studies on the Mechanism of Resistance to and Mode of Action of Viomycin in Mycobacterium smegmatis (Mycobacterium smegmatis를 이용한 Viomycin의 내성 및 작용 기전에 관한 연구)

  • 최응칠
    • YAKHAK HOEJI
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    • v.24 no.1
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    • pp.1-10
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    • 1980
  • Viomycin inhibited polypeptide biosynthesis, initiation complex formation and translocation of peptidyl-tRNA on ribosomes derived from a sensitive strain of Mycobacterium smegmatis (R-15), but not significantly on ribosomes from viomycin-resistant mutants(R-31 and R-43). The inhibition of translocation was stronger than that of initiation complex formation in the sensitive strain. The binding of [$^{14}C$] tuberactinomycin O, a viomycin analog, to ribosomal particles was studied by Millipore filter method. The sensitive ribosome exhibited higher affinity for the antibiotic than the resistant ribosomes. The resistance was localized on the large ribosomal subunit in a mutant(R-31), and on the small subunit in another mutant(R-43). The binding of the drug to the sensitive ribosomal subunit was markedly reduced by combination with the resistant pair subunit, and the entire ribosome became resistant to the antibiotic.

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Effect of γ-Irradiation on the Molecular Properties of Myoglobin

  • Lee, Yong-Woo;Song, Kyung-Bin
    • BMB Reports
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    • v.35 no.6
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    • pp.590-594
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    • 2002
  • To elucidate the effect of gamma-irradiation on the molecular properties of myoglobin, the secondary and tertiary structures, as well as the molecular weight size of the protein, were examined after irradiation at various irradiation doses. Gamma-irradiation of myoglobin solutions caused the disruption of the ordered structure of the protein molecules, as well as degradation, cross-linking, and aggregation of the polypeptide chains. A SDS-PAGE study indicated that irradiation caused initial fragmentation of the proteins and subsequent aggregation, due to cross-linking of the protein molecules. The effect of irradiation on the protein was more significant at lower protein concentrations. Ascorbic acid protected against the degradation and aggregation of proteins by scavenging oxygen radicals that are produced by irradiation. A circular dichroism study showed that an increase of the irradiation decreased the a-helical content of myoglobin with a concurrent increase of the aperiodic structure content. Fluorescence spectroscopy indicated that irradiation increased the emission intensity that was excited at 280 nm.

The Synthetic Potential of SET Photochemistry of Silicon-Substituted Polydonor-Linked Phthalimides

  • Yoon, Ung Chan;Mariano, Patrick S.
    • Bulletin of the Korean Chemical Society
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    • v.27 no.8
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    • pp.1099-1114
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    • 2006
  • Our studies in the area of single electron transfer (SET) photochemistry have led to the discovery of efficient processes, in which regioselective formation of carbon-centered radicals takes place by nucleophile assisted desilylation of $\alpha$-trialkylsilyl substituted ether, thioether, amine and amide centered cation radicals. The rates of bimolecular desilylation of the intermediate cation radicals exceed those of other cation radical $\alpha$-fragmentation processes (e.g.,-deprotonation). This sereves as the basis for the design of highly regioselective, SET-induced photomacrocyclization reactions of polyether, polythioether, polyamide, and polypeptide linked phthalimides. Photocyclization reactions of trimethylsilyl-terminated substrates in these families are unique in that they produce polyfunctionalized macrocyclic substances in a highly efficient and regioselective manner. In addition, our studies in this area have led to important information about the factors that govern chemical and quantum efficiencies that should be applicable to a wide variety of redox processes promoted by SET from substrates containing more than one electron donor site.