• Title/Summary/Keyword: pharmaceutical residues

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NMR Spectroscopic Assessment of the Structure and Dynamic Properties of an Amphibian Antimicrobial Peptide (Gaegurin 4) Bound to SDS Micelles

  • Park, Sang-Ho;Son, Woo-Sung;Kim, Yong-Jin;Kwon, Ae-Ran;Lee, Bong-Jin
    • BMB Reports
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    • v.40 no.2
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    • pp.261-269
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    • 2007
  • The structure and dynamics of a 37-residue antimicrobial peptide gaegurin 4 (GGN4) isolated from the skin of the native Korean frog, Rana rugosa, was determined in SDS micelles by NMR spectroscopy. The solution structure of the peptide in SDS micelles was determined from 352 NOE-derived distance constraints and 22 backbone torsion angle constraints. Dynamic properties for the amide backbone were characterized by $^1H-^{15}N $heteronuclear NOE experiments. The structural study revealed two amphipathic helices spanning residues 2-10 and 16-32 and that the helices were connected by a flexible loop. An intraresidue disulfide bridge was formed between residues Cys31 and Cys37 near the C-terminus. The loop region (11-15) connecting the two helices are were slightly more flexible than these helices themselves. From the fact that since there is no contact NOEs between two helices, it is implied that the GGN4 peptide shows an independent motion of both helices which has an angle of about $ 60^{\circ}-120^{\circ}$ from each other.

Backbone 1H, 15N, and 13C Resonance Assignment of HP1242 from Helicobacter pylori

  • Kang, Su-Jin;Park, Sung-Jean;Jung, Seo-Jeong;Lee, Bong-Jin
    • BMB Reports
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    • v.38 no.5
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    • pp.591-594
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    • 2005
  • One of the small proteins from Helicobacter pylori, HP1242, was investigated by the solution nuclear magnetic resonance (NMR) spectroscopy. HP1242 is known as a 76-residue conserved hypothetical protein and its function cannot be identified based on sequence homology. Here, the results of the backbone $^1H$, $^{15}N$, and $^{13}C$ resonance assignments of the HP1242 are reported using double- and triple-resonance techniques. About 95% of all of the $^1HN$, $^{15}N$, $^{13}CO$, $^{13}C{\alpha}$, and $^{13}C{\beta}$ resonances that cover 75 non- Proline residues of the 76 residues are clarified through sequential- and specific- assignments. In addition, three helical regions were clearly identified on the basis of the resonance assignments.

Pharmaceuticals in Environment and Their Implication in Environmental Health (의약물질의 환경오염과 환경보건)

  • Choi, Kyung-Ho;Kim, Pan-Gyi;Park, Jeong-Im
    • Journal of Environmental Health Sciences
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    • v.35 no.6
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    • pp.433-446
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    • 2009
  • Pharmaceuticals in the aquatic environment are trace contaminants of growing importance in environmental health due to their physiologically active nature. Pharmaceuticals could affect non-target species and might eventually damage sustainability of susceptible populations in the ecosystem. Potentials for health consequences among susceptible human population cannot be ruled out since long-term exposure to cocktails of pharmaceuticals, which might be present in drinking water, is possible. Selection of antibiotic resistant microorganisms is of another concern. In order to understand, and if needed, to properly address the environmental health issues of pharmaceutical residues, knowledge gaps need to be filled. Knowledge gaps exist in many important areas such as prioritization of target pharmaceuticals for further risk studies, occurrence patterns in different environments, chronic toxicities, and toxicities of pharmaceutical mixtures. Appropriate treatment technologies for drinking water and wastewater could be developed when they are deemed necessary. One of the simplest, yet most efficient measures that could be undertaken is to implement a return program for unused or expired drugs. In addition, implementation of environmental risk assessment frameworks for pharmaceuticals would make it possible to efficiently manage potential environmental health problems associated with pharmaceutical residues in the environment.

Bioactive Fabaceous Saponins and Structure-Activity Relationship

  • Kinjo, Junei;Nohara, Toshihiro
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.143-146
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    • 2002
  • Saponins are glycosidic compounds present in many edible and inedible plants. Structurally, they are composed of a lipid-soluble aglycone consisting of either a sterol or, more commonly, a triterpenoid and water-soluble sugar residues differing in type and amount of sugars [1]. Because of their amphiphilic nature, they are highly surface-active. Their biological activity is closely related to the chemical structures that determine the polarity, hydrophobicith and acidity of compounds [1]. (omitted)

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Expression and Cloning of Microbial Transglutaminase in S. cerevisiae (세균 유래 단백질연결효소 Transglutaminase의 클로닝과 효모에서의 발현)

  • Kim, Hyoun-Young;Oh, Dong-Soon;Kim, Jong-Hwa
    • The Korean Journal of Mycology
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    • v.36 no.1
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    • pp.93-97
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    • 2008
  • A $Ca^{2+}-independent$ microbial transglutaminase (mTGase) from the actinomycete Streptomyces mobaraensis IFO13819 is a useful enzyme in the food industry. It is consists 406 amino acid residues, which comprised leader and pro region of 75 amino acid residues and the structure region of 331 amino acid residues. Pro and structure gene of TGase were cloned into the yeast shuttle vector pYAEG-TER and then used to transform Saccharomyces cerevisiae 2805. Expression of mTGase in recombinant was confirmed with Northern hybridization and the maximal activity of TGase was shown 26 mU/ml.

Enhancement of 1,3-Dihydroxyacetone Production from Gluconobacter oxydans by Combined Mutagenesis

  • Lin, Xi;Liu, Sha;Xie, Guangrong;Chen, Jing;Li, Penghua;Chen, Jianhua
    • Journal of Microbiology and Biotechnology
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    • v.26 no.11
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    • pp.1908-1917
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    • 2016
  • Wild strain L-6 was subjected to combined mutagenesis, including UV irradiation, atmospheric and room temperature plasma, and ion beam implantation, to increase the yield of 1,3-dihydroxyacetone (DHA). With application of a high-throughput screening method, mutant Gluconobacter oxydans I-2-239 with a DHA productivity of 103.5 g/l in flask-shake fermentation was finally obtained with the starting glycerol concentration of 120 g/l, which was 115.7% higher than the wild strain. The cultivation time also decreased from 54 h to 36 h. Compared with the wild strain, a dramatic increase in enzyme activity was observed for the mutant strain, although the increase in biomass was limited. DNA and amino acid sequence alignment revealed 11 nucleotide substitutions and 10 amino acid substitutions between the sldAB of strains L-6 and I-2-239. Simulation of the 3-D structure and prediction of active site residues and PQQ binding site residues suggested that these mutations were mainly related to PQQ binding, which was speculated to be favorable for the catalyzing capacity of glycerol dehydrogenase. RT-qPCR assay indicated that the transcription levels of sldA and sldB in the mutant strain were respectively 4.8-fold and 5.4-fold higher than that in the wild strain, suggesting another possible reason for the increased DHA productivity of the mutant strain.

Role of Disulfide Bond of Arylsulfate Sulfotransferase in the Catalytic Activity

  • Kwon, Ae-Ran;Choi, Eung-Chil
    • Archives of Pharmacal Research
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    • v.28 no.5
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    • pp.561-565
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    • 2005
  • Bacterial arylsulfate sulfotransferase (ASST) catalyzes the transfer of sulfate group from a phenyl sulfate ester to a phenolic acceptor. The promoter region and the transcripti on start sites of Enterobacter amnigenus astA have been determined by primer extension analysis. Northern blot analysis resolved two mRNA species with lengths of 3.3 and 2.0 kb, which correspond to the distances between the transcriptional initiation sites and the two inverted repeat sequences (IRSs). By length, the 3.3 kb RNA could comprise the three-gene (astA with dsbA and dsbB) operon. ASST has three highly conserved cysteine residues. Reducing and non-reducing SDS-PAGE and activity staining showed that disulfide bond is needed for the activity of the enzyme. To identify the cysteine residues responsible for the disulfide bond formation, a series of Cys to Ser mutants has been constructed and the enzymatic activity was measured. Based on the results, we assumed that the first cysteine (Cys349) might be involved in disulfide bond mainly with the second cysteine (Cys445) and result in active conformation.

Cleaning Validation Studies for Multi-Purpose Facility : Vial Filling Machine (다품목 공용 제약설비인 바이알 충전기에 대한 세척공정 밸리데이션)

  • Choi, Han-Gon;Yang, Ho-Joon;Kim, Young-Ran;Sung, Jun-Ho;Hwang, Ma-Ro;Kim, Jong-Oh;Yong, Chul-Soon
    • Journal of Pharmaceutical Investigation
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    • v.39 no.4
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    • pp.263-267
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    • 2009
  • The purpose of this study is to evaluate the efficacy of stipulated cleaning process, and the prohibition of cross-contamination and microbiological contamination, which inadequate cleaning in multi-production could occur, through cleaning validation of multi-purpose facility used to produce five biopharmaceutical products as sterile injection. After production of five biopharmaceutical products such as hGH, rhGCSF, rhEPO, rhFSH and rhIFN using vial filling machine, the cleaning validation such as residual analysis of active ingredients or human serum albumin, measurement of total organic carbon (TOC), residual analysis of detergent and microbiological contamination were carried out. In the case of rhGH and rhGCSF clean validations, drug residues were not detected. Furthermore, in the case of rhEPO, rhFSH and rhIFN clean validations, human serum albumin residues were not detected. At TOC (total organic carbon) analysis, all clean validations gave the TOC of about average 137.93%, not more than 150% of acceptance criteria. At sodium analysis for the checking of residues of cleaning agent, sodium residues were not detected. In sterility test, they showed no microbiological contamination of bacteria and fungi. Thus, this cleaning validation was determined as successful in protection of cross-contamination and induction of safety in multi-purpose facility.