• 제목/요약/키워드: Bifunctional

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Production of Bioactive 3'-Hydroxystilbene Compounds Using the Flavin-Dependent Monooxygenase Sam5

  • Heo, Kyung Taek;Lee, Byeongsan;Son, Sangkeun;Ahn, Jong Seog;Jang, Jae-Hyuk;Hong, Young-Soo
    • Journal of Microbiology and Biotechnology
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    • 제28권7호
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    • pp.1105-1111
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    • 2018
  • The flavin-dependent monooxygenase Sam5 was previously reported to be a bifunctional hydroxylase with coumarate 3-hydroxylase and resveratrol 3'-hydroxylase activities. In this article, we showed the Sam5 enzyme has 3'-hydroxylation activities for methylated resveratrols (pinostilbene and pterostilbene), hydroxylated resveratrol (oxyresveratrol), and glycosylated resveratrol (piceid) as substrates. However, piceid, a glycone-type stilbene used as a substrate for bioconversion experiments with the Sam5 enzyme expressed in Escherichia coli, did not convert to the hydroxylated compound astringin, but it was converted by in vitro enzyme reactions. Finally, we report a novel catalytic activity of Sam5 monooxygenase for the synthesis of piceatannol derivatives, 3'-hydroxylated stilbene compounds. Development of this bioproduction method for the hydroxylation of stilbenes is challenging because of the difficulty in expressing P450-type hydroxylase in E. coli and regiospecific chemical synthesis.

Sodium 2-(2,3-dibromopropionylamino)-5-(4,6-dichloro-1,3,5-triazinylamino)-benzenesulfonate를 이용한 나일론/면 복합소재의 단일 산성염료 일욕염색 (Dyeing of Nylon/Cotton Blend with Acid Dyes Using Sodium 2-(2,3- dibromopropionylamino)-5-(4,6-dichloro-1,3,5-triazinylamino)-benzenesulfonate)

  • 김미경;윤남식;김태경
    • 한국염색가공학회지
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    • 제24권1호
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    • pp.8-17
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    • 2012
  • In order to dye nylon/cotton blended fabrics in solid colors with only acid dyes, a hetero-bifunctional bridge compound (DBDCBS) was examined. The bridge compound was designed to react only onto cotton fabrics first, on which amino-containing acid dyes react later by covalent bonding. By the effect of DBDCBS, amino-containing acid dyes exhibited high affinity toward the cotton fabrics. From the dyeing properties examined at various conditions, the optimum dyeing was decided at pH 4 and $100^{\circ}C$. Consequently, the solid color was obtained on nylon/cotton blended fabrics in one bath dyeing process using only acid dyes. The DBDCBS did not show any negative effect on nylon side in terms of dyeing and physical properties.

Photoinitiator-free Photo-reactive Coloration of Wool Fabrics Using C.I. Reactive Black 5

  • Dong, Yuanyuan;Jang, Jin-Ho
    • 한국염색가공학회지
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    • 제24권2호
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    • pp.97-105
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    • 2012
  • Compared with conventional adsorption-based coloration, the photoreactions of dyes such as photo-copolymerization and photo-crosslinking under UV irradiation can be employed for the coloration of textiles, which can be carried out without salt addition at room temperature. C.I. Reactive Black 5, a homo-bifunctional reactive dye containing two sulfatoethylsulfone groups, is used as a photo-reactive dye for wool fibers. Upon UV irradiation, the photo-reactive dye was grafted onto wool fabrics without photoinitiators. Since the disulfide bonds in the cystine residues of wool can be easily photodecomposed to active thiyl radicals which initiate the polymerization, the dye can be polymerized to an oligomeric dye of a degree of polymerization of 12 or more. The grafted fabrics reached a grafting yield of 2.3% o.w.f. and a color yield (K/S) of 18.2 by the photografting of an aqueous dye concentration of 9% using a UV energy of 25J/$cm^2$. Furthermore, the photochemically dyed wool fabric showed higher colorfastness properties to light, laundering and rubbing comparable to conventional reactive dyeing.

Catechol생산을 위한 Tyrosinase의 안정화 (Stabilization of Tyrosinase for Catechol Production)

  • 박종현;김용환유영제이윤식
    • KSBB Journal
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    • 제9권5호
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    • pp.525-531
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    • 1994
  • 본 연구에서는 tyrosinase 효소반응시에 발생하는 반응 비활성화를 막아 조엽 얀정성을 향상시키는 것을 목적으로 연구를 수행하였다. Quinone에 의해 친핵성 공격을 받는 lysine기를 bifunctional rea gent인 glutaraldehyde로 변형시켜 줌으로써 효소 의 비활성화를 줄일 수 있었다. 또한, 고정화된 효소 를 충진층 반응기에 충진시켜 비활생화를 열으키는 생산물을 연속적으로 제거함으로써 효소의 얀정성을 크게 향상시켰다. 고정화 방법으로는 glass bead를 이용한 담체 가교법을 이용하였으며, 이때 glutaral­d dehyde와 효소의 lysine이 반응하게 되어 qumone 에 의한 친핵성 공격을 받지 않게 되어 안정성이 더욱 향상되 었다. 반응매 질로 borate buffer를 이용하 게되면 catechol과 borate의 복합체 형성으로 catechol에서 qumone으로의 전환이 줄어들게 되어 효소 비활성화가 감소하였다.

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2,4-이니트로할로벤젠과 치환된 아닐린의 반응속도에 대한 용매효과 (제3보) (Influence of Solvents on Rates of Reactions of 2,4-Dinitro Substituted Halobenzenes with Substituted Anilines (III))

  • 이해황;이익춘
    • 대한화학회지
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    • 제22권4호
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    • pp.221-228
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    • 1978
  • 2,4-Dinitrohalo (F,Br) benzene과 파라치환아닐린간의 반응속도를 아세토니트릴-메탄올 혼합용매하에서 측정하였다. 2,4-Dinitrobromobenzene과 치환아닐간의 반응에서 메탄올은 친핵촉매 현상만을 보였으며, 2,4-dinitrofluorobenzene과 치환아닐린간의 반응에서는 메탄올은 양쪽성(친핵 및 친전자) 촉매현상을 보였다. 메탄올에 의한 친핵촉매현상은 아마도 천이상태에서 알코올기의 수소와 이탈기의 플루오르사이의 수소결합에 의해 설명될 수 있으며 친전자촉매현상은 알코올기의 산소와 아닐린의 아민기 수소사이의 수소결합에 기인되는 것임을 알았다.

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Novel Macromonomer as a Reactive Stabilizer in the Dispersion Polymerization of Methylmethacrylate

  • Jung, Hye-Jun;Lee, Kang-Seok;Shim, Sang-Eun;Yang, Sun-Hye;Lee, Jung-Min;Lee, Hui-Je;Choe, Soon-Ja
    • Macromolecular Research
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    • 제12권5호
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    • pp.512-518
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    • 2004
  • We have synthesized a novel macromonomer of vinyl-terminated bifunctional polyurethane having a molecular weight of 37,000 g/mol and successfully applied it to the dispersion polymerization of methylmethacrylate(MMA). We verified the presence of the vinyl terminal group and the macromonomer grafted onto the poly(ethylene glycol)(PEG) block in the PMMA particles by using $^1$H and $\^$13/C NMR spectroscopies. Monodisperse PMMA microspheres that have good uniformity of 1.01 were prepared at 20 wt% macromonomer content; we investigated the characteristics of the PMMA particles in terms of their molecular weight, molecular weight distribution, size of the particles, thermal properties, and glass transition temperature. We have found that the synthesized polyurethane macromonomer is an effective stabilizer.

Cofactor Regeneration Using Permeabilized Escherichia coli Expressing NAD(P)+-Dependent Glycerol-3-Phosphate Dehydrogenase

  • Rho, Ho Sik;Choi, Kyungoh
    • Journal of Microbiology and Biotechnology
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    • 제28권8호
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    • pp.1346-1351
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    • 2018
  • Oxidoreductases are effective biocatalysts, but their practical use is limited by the need for large quantities of NAD(P)H. In this study, a whole-cell biocatalyst for NAD(P)H cofactor regeneration was developed using the economical substrate glycerol. This cofactor regeneration system employs permeabilized Escherichia coli cells in which the glpD and gldA genes were deleted and the gpsA gene, which encodes $NAD(P)^+-dependent$ glycerol-3-phosphate dehydrogenase, was overexpressed. These manipulations were applied to block a side reaction (i.e., the conversion of glycerol to dihydroxyacetone) and to switch the glpD-encoding enzyme reaction to a gpsA-encoding enzyme reaction that generates both NADH and NADPH. We demonstrated the performance of the cofactor regeneration system using a lactate dehydrogenase reaction as a coupling reaction model. The developed biocatalyst involves an economical substrate, bifunctional regeneration of NAD(P)H, and simple reaction conditions as well as a stable environment for enzymes, and is thus applicable to a variety of oxidoreductase reactions requiring NAD(P)H regeneration.

Proton Transfer Reactions and Ion-Molecule Reactions of Ionized XCH2CH2Y (X and Y = OH or NH2)

  • Choi, Sung-Seen;So, Hun-Young
    • Bulletin of the Korean Chemical Society
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    • 제27권4호
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    • pp.539-544
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    • 2006
  • Proton transfer reactions and ion-molecule reactions of bifunctional ethanes of $H_2NCH_2CH_2NH_2$, $H_2NCH_2CH_2OH$, and $HOCH_2CH_2OH$ were studied using Fourier transform mass spectrometry (FTMS). The rate constants for proton transfer reactions between the fragment ions and neutral molecules were obtained from the temporal variation of the ion abundances. The rate constants were consistent with the heats of reaction. The fastest proton transfer reactions were the reactions of $CH_2N^+$, $CHO^+$, and $CH_3O^+$ for $H_2NCH_2CH_2NH_2$, $H_2NCH_2CH_2OH$, and $HOCH_2CH_2OH$, respectively. The $[M+13]^+$ ion was formed by the ion-molecule reaction between $H_2C=NH_2 ^+$ or $H_2C=OH^+$ and the neutral molecule. The major product ions generated from the ion-molecule reactions between the protonated molecule and neutral molecule were $[2M+H]^+$, $[M+27]^+$, and $[M+15]^+$.

Cloning and Characterization of Actinorhodin Biosynthetic Gene Clusters from Streptomyces lividans TK24

  • Park, Kie-In
    • Animal cells and systems
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    • 제6권4호
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    • pp.305-309
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    • 2002
  • Actinorhodin antibiotics produced by Streptomyces lividans TK24 are blue pigments with a weak antibiotic activity, derived from one acetyl-CoA and 15 malonyl-CoA units via a typical ployketide pathway. In an attempt to clone polyketide biosynthetic genes of S. lividans TK24, hybridizing fragments in the genomic DNA of S. lividans TK24 were detected by use of acn and act III polyketide synthase gene probes. Since typical aromatic polyketide bio-synthetic gene clusters are roughly 22-34 Kb long, we constructed in E. coli XL-Blue MR using the Streptomyces-E. coli bifunctional shuttle cosmid vector (pojn46). Then, about 5,000 individual E. coii colonies were thor-oughly screened with acrl-ORFI and actIII probes. From these cosmid libra-ries, 12 positive clones were identified. Restriction analysis and southern hybridization showed two polyketide biosynthetic gene clusters in this organism. These cosmid clones can be transformed into Streptomyces parvulus 12434 for expression test that identify product of actinorhodin biosynthetic genes by heterologous expression. Thus, heterologous expres-sion of a derivative compound of a actinorhodin biosynthetic intermediate was obtained in pKE2430. Expression of these compounds by the trans-formants was detected by photodiode array HPLC analysis of crude extracts.

A Comparative Analysis of Monofunctional Biosynthetic Peptidoglycan Transglycosylase (MBPT) from Pathogenic and Non-pathogenic Bacteria

  • Baker, Andrew T.;Takahashi, Natsumi;Chandra, Sathees B.
    • Genomics & Informatics
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    • 제8권2호
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    • pp.63-69
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    • 2010
  • Monofunctional biosynthetic peptidoglycan transglycosylase (MBPT) catalyzes the formation of the glycan chain in bacterial cell walls from peptidoglycan subunits: N-acetylglucosamine (NAG) and acetylmuramic acid (NAM). Bifunctional glycosyltransferases such as the penicillin binding protein (PBP) have peptidoglycan glycosyltransferase (PGT) on their C terminal end which links together the peptidoglycan subunits while transpeptidase (TP) on the N terminal end cross-links the peptide moieties on the NAM monosaccharide of the peptide subunits to create the bacterial cell wall. The singular function of MBPT resembles the C terminal end of PBP as it too contains and utilizes a similar PGT domain. In this article we analyzed the infectious and non infectious protein sequences of MBPT from 31 different strains of bacteria using a variety of bioinformatic tools. Motif analysis, dot-plot comparison, and phylogenetic analysis identified a number of significant differences between infectious and non-infectious protein sequences. In this paper we have made an attempt to explain, analyze and discuss these differences from an evolutionary perspective. The results of our sequence analysis may open the door for utilizing MBPT as a new target to fight a variety of infectious bacteria.