• Title/Summary/Keyword: Mutant

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Tetraacctylphytospingosinc (TAPS) 생산을 위한 Pichia ciferrii의 균주 개량과 TAPS 생산에 대한 sodium acetate의 영향

  • Hong, Seong-Gap;Yu, Yeon-U
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.370-373
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    • 2001
  • Experiments were carried out to develop of mutant strain from Pichia ciferrii ATCC 14091 and investigate the effect of sodium acetate on the production of tetraacetylphytosphingosine (TAPS). A high-TAPS producing mutant was simply selected by staining with sudan black B and colony shape after the treatment of UV and NTG. Sodium acetate enhanced the TAPS production.

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방선균의 xylB 변이주에 의한 포도당 이성화효소의 생산

  • 주길재;이인구
    • Microbiology and Biotechnology Letters
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    • v.25 no.1
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    • pp.75-81
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    • 1997
  • Streptomyces chibaensis J-59 did not grow in the culture medium containing only xylose or xylan as a carbon source, because it was defective in xylulokinase production; xylB mutant. S. chibaensis J-59 was able to produce xylanase and $\beta $-xylosidase as well as glucose isomerase. The glucose isomerase in S. chilbaensis J-59 was induced in the medium containing xylan or xylose which could be utilized as an inducer but not sa carbon and energy sources. So we tried to produce glucose isomerase whthout consumption of xylose or xylan as an inducer by using xylB mutant S. chilbaensis J-59. The optimum condition for the production of the glucose isomerase was attained in a culture medium composed of 1% xylan, 0.15% glucose, 1.5% corn steep liquor, 0.1% MaSO$_{4}$ $\CDOT $7H$_{2}$O, and 0.012% CoCL$_{2}$ $\CDOT $ 6H$_{2}$O(pH 7.0). The production of the enzyme reached to a maximum level when the bacteria were cultured for 42 h at 30$\circ $C. The enzyme production in a jar fermentor was increased twice as much as that in a flask culture.

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Performance Improvement of Genetic Algorithms by Strong Exploration and Strong Exploitation (감 탐색과 강 탐험에 의한 유전자 알고리즘의 성능 향상)

  • Jung, Sung-Hoon
    • Proceedings of the Korean Institute of Intelligent Systems Conference
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    • 2007.04a
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    • pp.233-236
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    • 2007
  • A new evolution method for strong exploration and strong exploitation termed queen-bee and mutant-bee evolution is proposed based on the previous queen-bee evolution [1]. Even though the queen-bee evolution has shown very good performances, two parameters for strong mutation are added to the genetic algorithms. This makes the application of genetic algorithms with queen-bee evolution difficult because the values of the two parameters are empirically decided by a trial-and-error method without a systematic method. The queen-bee and mutant-bee evolution has no this problem because it does not need additional parameters for strong mutation. Experimental results with typical problems showed that the queen-bee and mutant-bee evolution produced nearly similar results to the best ones of queen-bee evolution even though it didn't need to select proper values of additional parameters.

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Rapid Screening of Mutant Strains of Trigonopsis variabilis (ATCC10679) for Cephalosporin C Bioconversion and Sequences of D-amino acid oxidase Genes (Cephalosporin C 생물전환을 위한 Trigonopsis variabilis (ATCC10679) 변이균주의 간편한 선별 및 D-amino acid oxidase 유전자 배열)

  • 강용호;박선영
    • KSBB Journal
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    • v.14 no.2
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    • pp.235-240
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    • 1999
  • Simple and rapid screening methods were developed to screen mutant strains of Trigonopsis variabilis ATCC10679 (TW). D-amino acid oxidase (D-AAO) from a mutant strain, T26, showed about 30% higher specific activity against cephalosporin C than from its wild type, TW. D-AAO genes from both TW and T26 strains were cloned and sequenced. There was one nucleotide changed from T to C at 811 position, resulting in an amino acid codon changed from Phe-258 to Ser-258.

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Analysis of the Stoichiometry and the Domain for Interaction of Simian Virus 40 Small-t Antigen with Protein Phosphatase 2A

  • Yang, Sung-Il;Mumby, Marc C.
    • BMB Reports
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    • v.28 no.4
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    • pp.331-335
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    • 1995
  • Simian virus 40 (SV40) small-t antigen (small-t) has been known to regulate the activity of a cellular enzyme, protein phosphatase 2A (PP2A), composed of A. B, and C subunits, via binding to the A subunit In the study presented here, the stoichiometry of the binding of small-t to PP2A was determined to be 1: 1. It was also shown that small-t binds to the AC form of PP2A with a higher apparent affinity than it binds to the free A subunit. We also characterized the interaction of PP2A with wild-type and various mutant small-ts. A single-point mutant (Val134Met) and a double-point mutant (Trp147Gly;Leu152 Pro) of small-t exhibited 3-fold and 5-fold lower potencies in inhibiting PP2A activity. respectively. This suggests that the region around amino acids between 134 and 152 of small-t might be important in regulating the enzyme activity of PP2A.

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Proteomic Analysis of Circadian Clock Mutant Mice

  • Lee Joon-Woo;Kim Han-Gyu;Bae Kiho
    • Biomedical Science Letters
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    • v.11 no.4
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    • pp.493-501
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    • 2005
  • Circadian rhythms, time on a scale of about 24 hours, are present in a number of organisms including animals, plants, and bacteria. The control of the biochemical, physiological and behavioral processes is regulated by endogenous clocks in the suprachiasmatic nucleus (SCN). At the core of this timing mechanism is molecular machinery that are present both in the brain and in the peripheral tissues throughout the body, and even in a single cultured cell. In this study, we performed two-dimensional gel electrophoresis to figure out any correlation between protein expression patterns and the requirement of two canonical clock proteins, either mPER1 or mPER2, by comparing global protein expression profiles in livers from wildtype or mPer1/mPer2 double mutant mice. We could identify several differentially expressed protein candidates with respect to time and genotypes. Further analysis of these candidate proteins in detail in vivo will lead us to the better understanding of how circadian clock functions in mammals.

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Determinant Involved in the Loss of Pathogenicity in Wilt - Inducing Pseudomonas solanacerum (마름병 병원균 Pseudomonas solanacearum의 병원성 상실요인에 관하여)

  • 김을제;윤경란;이영하;이청호;박지창;최광태
    • Journal of the Korean Society of Tobacco Science
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    • v.12 no.1
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    • pp.9-18
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    • 1990
  • To study the determinants which are involved in the loss of pathogenicity in wilt-inducing Pseudomoms solamcewum, several physlologica I functions were compared in a virulent P. solanacearum strain and an avirulent, spontaneously derived mutant strain. The polyacrylamide gel electrophoresis showed the distinction between two strains in the patterns and the relative intensity of proteins produced intracellularly or extracellularly. Enzyme assays showed that the level of polygalacturonase activity in the culture filtrate of the avirulent mutant was markedly reduced, while carboxymethylcellulase(rondoglucanase) activity in both strains were nearly negligible. These results suggest that the loss of pathogenicity in mutant strain is attributed in part to the reduced production of polygalacturonase. In audit ion, comparative analyses by agarose gel electrophoresis of DNA molecules isolated from both strains show that the pathogenicity genes of p. solanaceerum are not located on plasmid but are on chromosome.

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Subcellular Localization of Catalase Encoded by the ctl+ Gene in Schizosaccharomyces pombe

  • Lee, Sang-il;Lee, Joon;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.38 no.3
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    • pp.156-159
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    • 2000
  • The cttl+ gene in Schizosaccharomyces pombe encoeds a catalse responsible for H2O2-resistance of this organism as judged by the H2O2-sensitive phenotype of the ctt1Δ mutant. In this study, we investigated the subcellular localization of the Ctt1 gene product. In wild type cells catalase activity was detected in the organelle fraction as well as in the cytosol. The ctt1Δ mutant contained no catalase activity, indicating that both cytosolic and organellar catalases are the products of a single ctt1+ gene. Western bolt analysis revealed two catalase bands, both of which disappeared in the ctt1Δ mutant. The major, fastermigrating band existed in the cytosol whereas the monor, slower-migrating band appeared to be located in organelles, most likely in peroxisomes. These results suggest that the ctt1+ gene product targeted to the peroxisome is a modified form of the one in the cytosol.

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Abscisic Acid Binding to Extracts from Normal and Viviparous-1 Mutant Aleurone Layers of Zea mays L.

  • Bai, Dong-Gyu
    • Journal of Plant Biology
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    • v.37 no.2
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    • pp.151-158
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    • 1994
  • Aleurone layers of normal and vp1 mutant maize kernels were extracted and centrifuged at 100,000g to yield a cytosol fraction. Binding of [3H]ABA cis, trans (+)ABA to a soluble macromolecular components present in the cytosol was demonstrated by Sephadex chromatography and non-denaturing PAGE. The binding component was of high molecular weight and seems to be an aggregate of proteins. A rapid DEAE-cellulose filter method for assaying bound [3H]ABA to a soluble protein was adapted. Binding assays were performed with cytosol that had been preheated or incubated with several enzymes, indicating that heat and protease treatments disrupted the binding. This suggested that binding occurred to proteins. Some properties of the ABA binding proteins were described. The [3H]ABA binding were reduced dramatically when unlabeled ABA was added as a competitor, suggesting a specific binding of [3H]ABA. Gel filtration profiles and autoradiogram of [3H]ABA binding showed no difference in the binding components of Vp1 and vp1/vp1 mutant cytosol, indicating that Vp1 protein is not a sole ABA binding protein.

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The effect of surface charge balance on thermodynamic stability and kinetics of refolding of firefly luciferase

  • Khalifeh, Khosrow;Ranjbar, Bijan;Alipour, Bagher Said;Hosseinkhani, Saman
    • BMB Reports
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    • v.44 no.2
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    • pp.102-106
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    • 2011
  • Thermodynamic stability and refolding kinetics of firefly luciferase and three representative mutants with depletion of negative charge on a flexible loop via substitution of Glu by Arg (ER mutant) or Lys (EK mutant) as well as insertion of another Arg in ER mutants (ERR mutant) was investigated. According to thermodynamic studies, structural stability of ERR and ER mutants are enhanced compared to WT protein, whereas, these mutants become prone to aggregation at higher temperatures. Accordingly, it was concluded that enhanced structural stability of mutants depends on more compactness of folded state, whereas aggregation at higher temperatures in mutants is due to weakening of intermolecular repulsive electrostatic interactions and increase of intermolecular hydrophobic interactions. Kinetic results indicate that early events of protein folding are accelerated in mutants.