• 제목/요약/키워드: Substitution site

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Functional Studies of Cysteine Residues in Human Glutathione S-Transferase P1-1 by Site-Directed Mutagenesis

  • Park, Hui Jung;Lee, Gwang Su;Gong, Gwang Hun
    • Bulletin of the Korean Chemical Society
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    • v.22 no.1
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    • pp.77-83
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    • 2001
  • To gain further insight into the relationship between structure and function of glutathione S-transferase (GST), the four cysteine mutants, C14S, C47S, C101S and C169S, of human GST P1-1 were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized glutathione (GSH). The catalytic activities of the four mutant enzymes were characterized with five different substrates as well as by their binding to four different inhibitors. Cys14 seems to participate in the catalytic reaction of GST by stabilizing the conformation of the active-site loop, not in the GSH binding directly. The substitution of Cys47 with serine significantly reduces the affinity of GSH binding, although it does not prevent GSH binding. On the other hand, the substitution of Cys101 with serine appears to change the binding affinity of electrophilic substrate by inducing a conformational change of the $\alpha-helix$ D. Cys169 seems to be important for maintaining the stable conformation of the enzyme. In addition, all four cysteine residues are not needed for the steroid isomerase activity of human glutathione S-transferase P1-1.

Local Structure Refinement of the $BaFe_{1-x}Sn_xO_{3-y}$ System with Fe K-Edge X-Ray Absorption (XANES/EXAFS) Spectroscopy

  • 김민규;곽기섭;로권선;여철현
    • Bulletin of the Korean Chemical Society
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    • v.18 no.7
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    • pp.743-749
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    • 1997
  • Local structure refinement of the BaFe1-xSnxO3-y system (x=0.00-0.50) has been carried out with Fe K-edge x-ray absorpion spectroscopic studies. It is found out that the Fe ions are placed in two different symmetric sites such as tetrahedral and octahedral sites in the compounds by comparison with Fe K-edge x-ray absorption near edge structure (XANES) spectrum of the γ-Fe2O3 compound as a reference. Small absorption peaks of dipole-forbiden transitions appear at a pre-edge region of 7111 eV due to the existence of Fe ions in the tetrahedral and octahedral sites. The peak intensity decreases with the substitution amount of Sn ion. Three different absorption peaks of 1s→4p dipole-allowed transition appear on the energy region between 7123 and 7131 eV. The peaks correspond to 1s→4p main transition of Fe ions in tetrahedral and octahedral sites and 1s→4p transition followed by the shakedown process of ligand to metal charge transfer. The bond distances between Fe ions in the tetrahedral site and nearest neighboring oxygen atom (Fe-4O), and those in octahedral site (Fe-6O) are determined with the extended x-ray absorption fine structure (EXAFS) analysis. Two different interatomic distances increase with the substitution amount of Sn ion and also the bond lengths of Fe-4O are shorter than those of Fe-6O in all compounds.

Computational screening of electroactive indolequinone derivatives as high-performance active materials for aqueous redox flow batteries

  • Han, Young-Kyu;Jin, Chang-Soo
    • Current Applied Physics
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    • v.18 no.12
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    • pp.1507-1512
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    • 2018
  • The development of an organic-based aqueous redox flow battery (RFB) using quinone as an electroactive material has attracted great attention recently. This is because this battery is inexpensive, produces high energy density, and is environment friendly in stationary electrical energy storage applications. Herein, we investigate the redox potentials and solubilities of indole-5,6-quinone and indole-4,7-quinone derivatives in terms of the substituent effects of functional groups using theoretical calculations. Our results indicate that full-site substituted derivatives of indolequinone are more useful as active materials compared to single-site substituted derivatives. In particular, our calculations reveal that the substitution of $-PO_3H_2$ and $-SO_3H$ functional groups with multiple polar bonds is very effective in increasing the activity of the aqueous RFB. As a strategy to overcome the limitation that the aqueous solubility is intrinsically low because they are organic molecules, we suggest the substitution of functional groups with multiple polar bonds to the backbones of active organic materials. Among 180 indolequinone derivatives, 17 candidates that meet the redox potential standards ($${\leq_-}0.2V$$ or $${\geq_-}0.9V$$) and eight candidates with solubility exceeding 2 mol/L are identified. Three indolequinone derivatives that satisfy both conditions are finally presented as promising electroactive candidates for an aqueous RFB.

Improvement of Bacilysin Production in Bacillus subtilis by CRISPR/Cas9-Mediated Editing of the 5'-Untranslated Region of the bac Operon

  • Hadeel Waleed Abdulmalek;Ayten Yazgan-Karatas
    • Journal of Microbiology and Biotechnology
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    • v.33 no.3
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    • pp.410-418
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    • 2023
  • Bacilysin is a dipeptide antibiotic composed of L-alanine and L-anticapsin produced by certain strains of Bacillus subtilis. Bacilysin is gaining increasing attention in industrial agriculture and pharmaceutical industries due to its potent antagonistic effects on various bacterial, fungal, and algal pathogens. However, its use in industrial applications is hindered by its low production in the native producer. The biosynthesis of bacilysin is mainly based on the bacABCDEF operon. Examination of the sequence surrounding the upstream of the bac operon did not reveal a clear, strong ribosome binding site (RBS). Therefore, in this study, we aimed to investigate the impact of RBS as a potential route to improve bacilysin production. For this, the 5' untranslated region (5'UTR) of the bac operon was edited using the CRISPR/Cas9 approach by introducing a strong ribosome binding sequence carrying the canonical Shine-Dalgarno sequence (TAAGGAGG) with an 8 nt spacing from the AUG start codon. Strong RBS substitution resulted in a 2.87-fold increase in bacilysin production without affecting growth. Strong RBS substitution also improved the mRNA stability of the bac operon. All these data revealed that extensive RBS engineering is a promising key option for enhancing bacilysin production in its native producers.

The Study of Sintering Behavior and Piezoelectric Properties in $Pd_{(1-x)}$$Cd_x$[(Mn, Sb), Zr, $Ti]O_3$ Ceramics ($Pd_{(1-x)}$$Cd_x$[(Mn, Sb), Zr, $Ti]O_3$ 세라믹스의 소결 거동 및 압전 특성에 대한 연구)

  • 나은상;최성철
    • Journal of the Korean Ceramic Society
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    • v.37 no.4
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    • pp.395-401
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    • 2000
  • In this study, we chose the basic composition which indicated the best electrical properties by change of x content(0, 0.05, 0.1, 0.15, 0.2, 0.25 mol respectively) in xPb(Mn1/3Sb2/3)O3-(1-x)Pb(Zr0.52Ti0.48)O3 ceramics. And we substituted Cd2+ for Pb2+ site, then observed the sintering behavior, microstructure and electrical propertties according to the various sintering temperature. The basic composition was the 0.05PMS-0.95PZt, and it showed single perovskite phase and excellent properties. In case of Cd2+ substitution, we were able to sinter at 90$0^{\circ}C$ which was lower than conventional sintering temperature(1200~130$0^{\circ}C$). Especially, when the 2mol% substituted PMS-PZT specimens were sintered at 90$0^{\circ}C$ for 2h, we obtained the p=7.6g/㎤, kp=56%, Qm=520 and made sure of a position of Cd2+ substitution by observing lattice parameter, phase transition temperature. From this results, we could infer that because Cd2+ substituted fro A-site, low temperature sintering of Cd2+ substituted PMS-PZT without any loss of electrical properties shows its applicability for the piezoelectric ceramic transformer.

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Biochemical Properties of Second Site Mutation of Human Immunodeficiency Virus Integrase

  • Kim, Do-Jin;Oh, You-Take;Lee, Sang-Kwang;Shin, Cha-Gyun
    • BMB Reports
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    • v.32 no.6
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    • pp.599-604
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    • 1999
  • A highly conserved amino acid, glutamic acid (Glu), present at position 152 in the catalytic domain of the human immunodeficiency virus type 1 (HIV-1) integrase (IN) protein has been known to be critical for enzymatic function since substitution of Glu 152 with other residues results in a complete loss of enzymatic activities. In order to better understand the role of Glu 152 as a conserved residue in enzymatic action, intragenic second site mutations have been introduced around residue 152 of a mutant IN (E152A), and their biochemical properties were analyzed in terms of enzymatic activities. Disintegration activities were found to be significantly restored in several second site mutant INs, while integration activities were only recovered weakly. However, endonucleolytic activities were not discovered in all the mutant INs. These findings indicate that the second site mutations can partially restore that catalytic structure of the active site disturbed by the E152A mutation and lead to the regaining of integration and disintegration activities. In addition, it is also suggested that endonucleolytic activity requires a more accurate structure of the catalytic site than that for the integration and disintegration activities.

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Effect of Gd Substitution for the Ca Site in the Bi1.84Pb0.34Sr1.91 ({Ca1-xGdx)2.03Cu3.06O10+δ(x=0.0~0.06) Superconductors

  • Lee, Min-Soo
    • Journal of the Korean Ceramic Society
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    • v.40 no.5
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    • pp.405-409
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    • 2003
  • The effect of substitution of Gd ions for Ca ions in the B $i_{1.84}$P $b_{0.34}$S $r_{1.91}$(C $a_{1-x}$ G $d_{x}$)$_{2.03}$ C $u_{3.06}$ $O_{ 10+{\delta}}$/ (x=0.0~0.06) was investigated by measuring x-ray diffraction patterns, lattice constants, do resistivity and Hall effect. We found the solubility limit of Gd in the 110 K phase to be x < 0.015. Within the solubility limit, the c-axis seemed to decrease with increasing x. In the region of the 110 K single phase, the critical temperature $T_{c}$ gradually decreased with an increasing the Gd concentration x, corresponding to a small change of the carrier concentration.

Catalytic Properties of Borosilicate in Methanol Conversion (메탄올의 전환반응에서 보로실리케이트의 촉매성질)

  • Lee, Gye Su;Jo, Min Su;Jeong, Byeong Gu;Seo, Gon
    • Journal of the Korean Chemical Society
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    • v.34 no.4
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    • pp.360-369
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    • 1990
  • Borosilicate, HZSM-5 zeolite and iron-substituted borosilicate and HZSM-5 zeolite were prepared and their catalytic properties in methanol conversion were studied. The effects of strength and amount of acid site determined from TPD spectra of ammonia on the product distribution was examined. Selectivity to propylene was high over borosilicate with small amount of strong acid site, but selectivity to aromatic compound was high over HZSM-5 zeolite with large amount of the strong acid site. The participation of weak acid site on the conversion did not confirmed, and the product distribution could be explained in terms of the amount of the strong acid site. Although the amount of the weak acid site was increased by substitution of iron, there was no meaningful change in the product distribution.

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Effect of Polar Amino Acid Residue Substitution by Site-Directed Mutagenesis in the N-terminal Domain of Pseudomonas sp. Phytase on Enzyme Activity

  • Lee, Ga Hye;Jang, Won Je;Kim, Soyeong;Kim, Yoonha;Kong, In-Soo
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.1104-1107
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    • 2020
  • The N-terminal domain of the Pseudomonas sp. FB15 phytase increases low-temperature activity and catalytic efficiency. In this study, the 3D structure of the N-terminal domain was predicted and substitutions for the amino acid residues of the region assumed to be the active site were made. The activity of mutants, in which alanine (A) was substituted for the original residue, was investigated at various temperatures and pH values. Significant differences in enzymatic activity were observed only in mutant E263A, suggesting that the amino acid residue at position 263 of the N-terminal domain is important in enzyme activity.

Sequence Analysis of the Internal Transcribed Spacer of Ribosomal DNA in the Genus Rhizopus

  • Park, You-Jung;Min, Byung-Re
    • Mycobiology
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    • v.33 no.2
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    • pp.109-112
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    • 2005
  • The internal transcribed spacer (ITS) regions including the 3'-end of 18S rRNA gene, 5.8S rRNA gene and the 5'-end of the 28S rRNA gene of Rhizopus spp. were amplified by PCR and analyzed by DNASIS program. Length polymorphism of these region ranged from 564 bp in R. oryzae to 789bp in R. stolonifer. The length and sequence of 5.8S was very conserved with $154{\sim}155\;bp$. The sequence of ITS2 was more variable than that of ITS1. The base substitution rates were ranged from 0 to 0.6069 per site, and higher rate was found in R. stolonifer. In general, transition was usually more frequent than transversion. On the basis of sequencing results, four groups were clustered with value of 61.9% similarity; R. oryzae, R. micros pores, R. homothallicus, and R. stolonifer groups.