• 제목/요약/키워드: acceptor reaction

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김치에서 분리한 Bacillus sp. S-6의 Cyclodextrin Glucanotransferase의 특성과 최적생산조건 (Some Properties and Optimal Culture Conditions of Cyclodextrin Glucanotransferase of Bacillus sp. S-6 Isolated from Kimchi)

  • 전홍기;조영배;김수진;배경미
    • 한국식품영양과학회지
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    • 제27권4호
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    • pp.609-617
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    • 1998
  • A microorganism capable of producing high level of extracellular cyclodextrin glucanotransferase(EC 2.4.1.19 ; CGTase) was isolated from Kimchi. 2-O-$\alpha$-D-glucopyranosyl L-ascorbic acid(AA-2G) was synthesized by transglycosylation reaction of CGTase using starch as a donor and L-ascorbic acid as an acceptor. The isolated strain S-6 was identified as Bacillus sp. S-6. The maximal CGTase production was observed in a medium containing 0.5% soluble starch, 1% yeast extract, 1% NaCO3, 0.1% K2HPO4, and 0.02% MgSO4 with initial pH 8.0. The strain was cultured at 37$^{\circ}C$ for 40 hr with reciprocal shaking. Using the culture supernatant as crude enzyme, the optimal pH and temperature of the CGTase activity of this strain were 7.0 and 4$0^{\circ}C$. In the effects of pH and temperature on the stability of the enzyme, the enzyme was stable in the range of pH 6.0~10.0 and up to 45$^{\circ}C$, respectively.

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Activity Determination, Kinetic Analyses and Isoenzyme Identification of Gamma Glutamyltransferase in Human Neutrophils

  • Sener, Azize;Yardimci, Turay
    • BMB Reports
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    • 제38권3호
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    • pp.343-349
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    • 2005
  • Gamma-glutamyltransferase (GGT, EC 2.3.2.2) which hydrolyzes glutathione (GSH), is required for the maintenance of normal intracellular GSH concentration. GGT is a membrane enzyme present in leukocytes and platelets. Its activity has also been observed in human neutrophils. In this study, GGT was purified from Triton X-100 solubilized neutrophils and its kinetic parameters were determined. For kinetic analyses of transpeptidation reaction, $\gamma$-glutamyl p-nitroanilide was used as the substrate and glycylglycine as the acceptor. Apparent $K_m$ values were determined as 1.8 mM for $\gamma$-glutamyl p-nitroanilide and 16.9 mM for glycylglycine. The optimum pH of GGT activity was 8.2 and the optimum temperature was $37^{\circ}C$. It had thermal stability with 58% relative activity at $56^{\circ}C$ for 30 min incubation. L-serine, in the presence of borate, was detected as the competetive inhibitor. Bromcresol green inhibited neutrophil GGT activity as a noncompetetive inhibitor. The neutrophils seem to contain only the isoenzyme that is present in platelets. We characterized the kinetic properties and compared the type of the isoenzyme of neutrophil GGT with platelet GGT via polyacrylamide gel electrophoresis (PAGE) under a standart set of conditions.

Substituent Effects on the Gas-Phase Pyrolyses of 2-Substituted Ethyl N,N-Dialkylcarbamates: A Theoretical Study

  • Kim, Chang-Kon;Kim, Dong-Jin;Zhang, Hui;Hsieh, Yih-huang;Lee, Bon-Su;Lee, Hai-Whang;Kim, Chan-Kyung
    • Bulletin of the Korean Chemical Society
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    • 제28권6호
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    • pp.1031-1034
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    • 2007
  • The R- and Z-substituent effects for the gas-phase thermal decompositions of carbamates, R2NC(=O)- OCH2CH2Z, have been investigated theoretically at B3LYP level with 6-31G(d) and 6-31++G(d,p) basis sets. Both the Z- and R-substituent effects on reactivity (ΔH≠) were well consistent with experimental results, although the R-substituent effect was underestimated theoretically. No correlations were found between activation enthalpies and reaction enthalpies. The substituent effects on reactivity seemed to be complicated at a glance, but were understandable by concurrent electronic and steric factors. Variations of bond lengths at TS structures were well correlated with the Taft's σ* values and the TS structures became tighter as the Zsubstituent became a stronger electron-acceptor (δσ* > 0). However the effects of R-substituents on the TS structures were much smaller when compared to those of Z-substituents.

Crystallization and Preliminary X-Ray Diffraction Analysis of BcOMT2 from Bacillus cereus: A Family of O-Methyltransferase

  • Cho, Jang-Hee;Lim, Yoong-Ho;Ahn, Joong-Hoon;Rhee, Sang-Kee
    • Journal of Microbiology and Biotechnology
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    • 제17권2호
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    • pp.369-372
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    • 2007
  • O-Methyltransferases (OMTs), one of the ubiquitous enzymes in plants, bacteria, and humans, catalyze a methyl-transfer reaction using S-adenosylmethionine and a wide range of phenolics as a methyl donor and acceptor, respectively. Substrates for most bacterial OMTs have largely remained elusive, but recent investigation using BcOMT2, an OMT from Bacillus cereus, suggested that ortho-dihydroxyflavonoids could serve as substrates. To elucidate the functional and structural features of BcOMT2, we expressed, and purified BcOMT2, and crystallized an apoenzyme and its ternary complex in the presence of a flavonoid and S-adenosylhomocysteine. Each crystal diffracted to $1.8{\AA}$ with its space group of C2 and $P2_{1}2_{1}2_{1}$, respectively. Structural analysis of apo-BcOMT2 and its ternary complex will provide the structural basis of methyl transfer onto (iso)flavonoids in a regiospecific manner.

탈질조건에서 nitroglycerin의 생물학적 분해 동역학 및 미생물 군집 변화 (Nitroglycerin Biodegradation under Denitrification Conditions and Corresponding Microbial Community Shifts upon Acclimation)

  • 최원철;배범한
    • 한국지하수토양환경학회지:지하수토양환경
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    • 제24권5호
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    • pp.42-54
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    • 2019
  • Biodegradation of an explosive compound, glyceryl trinitrate (GTN), was studied with a denitrifying microbial culture grown in a sequencing batch reactor and a GTN acclimated denitrifying culture. The GTN acclimated culture, which were fed on GTN for 1 month, degraded GTN regioselectively via denitration on C1 position as compared to C2 position denitration by denitrifying culture that has never been exposed to GTN. Accumulation of two isomeric glyceryl dinitrates (GDNs) in both culture medium suggests that GDN denitration is the rate-limiting step in GTN biodegradation. The first order GTN degradation rate normalized to cell concentration of the acclimated culture was calculated to be 0.045 (${\pm}0.002$) L/g-hr. Increasing concentration of electron acceptor(nitrate) resulted in discouraged GTN degradation. According to microbial community analysis, prolonged GTN exposure resulted in 25% increase in the genus level of the GTN acclimated culture with the disappearance of two dominating denitrifying microbial species of Methyloversatilis universalis and Hyphomicrobium zavarzinii in the denitrifying culture.

유기발광 디바이스용 녹색 발광재료의 합성 (Synthesis of Green Emitting Materials for OLED)

  • 정평진;김미래
    • 공업화학
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    • 제22권6호
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    • pp.594-598
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    • 2011
  • 본 연구는 유기발광 디바이스용(Organic Light Emitting Device, OLED) 녹색 발광재료인 3-크로몬알데히드 유도체의 합성에 관한 것으로서, 유도체들은 탈수 축합반응으로 합성되었다. 이들은 전자흡인성의 3-크로몬알데히드류와 전자공여성의 디아민류의 공액구조를 가지고 있다. 합성한 물질들은 각각 FT-IR, $^1H-NMR$ 스펙트럼으로부터 그의 구조적 특성을 확인하였고, 융점, 수득률 등을 통하여 열적 안정성, 반응성들을 확인하였으며, 여기스펙트럼과 발광스펙트럼으로부터 자외가시광과 발광특성을 확인하였다.

Glycosylation of Semi-Synthetic Isoflavene Phenoxodiol with a Recombinant Glycosyltransferase from Micromonospora echinospora ATCC 27932

  • Seo, Minsuk;Seol, Yurin;Park, Je Won
    • Journal of Microbiology and Biotechnology
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    • 제32권5호
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    • pp.657-662
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    • 2022
  • Glycosyltransferase (GT)-specific degenerate PCR screening followed by in silico sequence analyses of the target clone was used to isolate a member of family1 GT-encoding genes from the established fosmid libraries of soil actinomycetes Micromonospora echinospora ATCC 27932. A recombinant MeUGT1 was heterologously expressed as a His-tagged protein in E. coli, and its enzymatic reaction with semi-synthetic phenoxodiol isoflavene (as a glycosyl acceptor) and uridine diphosphate-glucose (as a glycosyl donor) created two different glycol-attached products, thus revealing that MeUGT1 functions as an isoflavonoid glycosyltransferase with regional flexibility. Chromatographic separation of product glycosides followed by the instrumental analyses, clearly confirmed these previously unprecedented glycosides as phenoxodiol-4'-α-O-glucoside and phenoxodiol-7-α-O-glucoside, respectively. The antioxidant activities of the above glycosides are almost the same as that of parental phenoxodiol, whereas their anti-proliferative activities are all superior to that of cisplatin (the most common platinum chemotherapy drug) against two human carcinoma cells, ovarian SKOV-3 and prostate DU-145. In addition, they are more water-soluble than their parental aglycone, as well as remaining intractable to the simulated in vitro digestion test, hence demonstrating the pharmacological potential for the enhanced bio-accessibility of phenoxodiol glycosides. This is the first report on the microbial enzymatic biosynthesis of phenoxodiol glucosides.

Influence of Reactive Media Composition and Chemical Oxygen Demand as Methanol on Autotrophic Sulfur Denitrification

  • Qambrani, Naveed Ahmed;Oh, Sang-Eun
    • Journal of Microbiology and Biotechnology
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    • 제22권8호
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    • pp.1155-1160
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    • 2012
  • Sulfur-utilizing autotrophic denitrification relies on an inorganic carbon source to reduce the nitrate by producing sulfuric acid as an end product and can be used for the treatment of wastewaters containing high levels of nitrates. In this study, sulfur-denitrifying bacteria were used in anoxic batch tests with sulfur as the electron donor and nitrate as the electron acceptor. Various medium components were tested under different conditions. Sulfur denitrification can drop the medium pH by producing acid, thus stopping the process half way. To control this mechanism, a 2:1 ratio of sulfur to oyster shell powder was used. Oyster shell powder addition to a sulfur-denitrifying reactor completely removed the nitrate. Using 50, 100, and 200 g of sulfur particles, reaction rate constants of 5.33, 6.29, and $7.96mg^{1/2}/l^{1/2}{\cdot}h$ were obtained, respectively; and using 200 g of sulfur particles showed the highest nitrate removal rates. For different sulfur particle sizes ranging from small (0.85-2.0 mm), medium (2.0-4.0 mm), and large (4.0-4.75 mm), reaction rate constants of 31.56, 10.88, and $6.23mg^{1/2}/l^{1/2}{\cdot}h$ were calculated. The fastest nitrate removal rate was observed for the smallest particle size. Addition of chemical oxygen demand (COD), methanol as the external carbon source, with the autotrophic denitrification in sufficiently alkaline conditions, created a balance between heterotrophic denitrification (which raises the pH) and sulfur-utilizing autotrophic denitrification, which lowers the pH.

Enzymatic Characterization and Substrate Specificity of Thermostable $\beta-Glycosidase$ from Hyperthermophilic Archaea, Sulfolobus shibatae, Expressed in E. coli

  • Park, Na-Young;Cha, Jae-Ho;Kim, Dae-Ok;Park, Cheon-Seok
    • Journal of Microbiology and Biotechnology
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    • 제17권3호
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    • pp.454-460
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    • 2007
  • Enzymatic properties and substrate specificity of recombinant $\beta-glycosidases$ from a hyperthermophilic archaeon, Sulfolobus shibatae (rSSG), were analyzed. rSSG showed its optimum temperature and pH at $95^{\circ}C$ and pH 5.0, respectively. Thermal inactivation of rSSG showed that its half-life of enzymatic activity at $75^{\circ}C$ was 15 h whereas it drastically decreased to 3.9 min at $95^{\circ}C$. The addition of 10 mM of $MnCl_2$ enhanced the hydrolysis activity of rSSG up to 23% whereas most metal ions did not show any considerable effect. Dithiothreitol (DTT) and 2-mercaptoethanol exhibited significant influence on the increase of the hydrolysis activity of rSSG rSSG apparently preferred laminaribiose $(\beta1\rightarrow3Glc)$, followed by sophorose $(\beta1\rightarrow2Glc)$, gentiobiose $(\beta1\rightarrow6Glc)$, and cellobiose $(\beta1\rightarrow4Glc)$. Various. intermolecular transfer products were formed by rSSG in the lactose reaction, indicating that rSSG prefers lactose as a good acceptor as well as a donor. The strong intermolecular transglycosylation activity of rSSG can be applied in making functional oligosaccharides.

$\alpha$-Cyano-$\beta$-piperonylacrylic Acid에 대한 n-Propylmercaptan의 친핵성 첨가반응에 관한 연구 (The Kinetics and Mechanism of Nucleophilic Addition of n-Propylmercaptan to $\alpha$-Cyano-$\beta$-piperonylacrylic Acid)

  • 김태린;허태성
    • 대한화학회지
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    • 제17권5호
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    • pp.363-370
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    • 1973
  • $\alpha$-Cyano-$\beta$-piperonylacrylic acid에 대한 n-Propylmercaptan의 친핵성 첨가반응 속도 상수를 측정하여 넓은 pH 범위에서 잘 맞는 반응속도식을 구하였다. 이 식에 의하면 넓은 pH 범위에서 일어나는 복잡한 반응메카니즘을 정량적으로 잘 설명할 수 있었다. 즉 pH4.5이하에서는 $\alpha$-Cyano-$\beta$-piperonylacrylic acid에 mercaptan이 직접 첨가되며 pH 4.5∼6.5 사이에서는 주로 $\alpha$-Cyano-$\beta$-piperonylacrylic ion에 mercaptan 분자가 첨가되고 pH 7.5 부터는 $\alpha$-Cyano-$\beta$-piperonylacrylic ion에 mercaptan 분자와 mercaptide ion의 첨가가 경쟁적으로 일어나며 pH 10이상에서는 $\alpha$-Cyano-$\beta$-piperonylacrylic anion에 mercaptide ion의 첨가만이 일어남을 알 수 있었다.

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