• Title/Summary/Keyword: Artificial enzyme

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Purification and Characterization of Manganese Peroxidase of the White-Rot Fungus Irpex lacteus

  • Shin Kwang-Soo;Kim Young Hwan;Lim Jong-Soon
    • Journal of Microbiology
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    • v.43 no.6
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    • pp.503-509
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    • 2005
  • The production of manganese peroxidase (MnP) by Irpex lacteus, purified to electrophoretic homogeneity by acetone precipitation, HiPrep Q and HiPrep Sephacryl S-200 chromatography, was shown to correlate with the decolorization of textile industry wastewater. The MnP was purified 11.0-fold, with an overall yield of $24.3\%$. The molecular mass of the native enzyme, as determined by gel filtration chromatography, was about 53 kDa. The enzyme was shown to have a molecular mass of 53.2 and 38.3 kDa on SDS-PAGE and MALDI-TOF mass spectrometry, respectively, and an isoelectric point of about 3.7. The enzyme was optimally active at pH 6.0 and between 30 and $40^{\circ}C$. The enzyme efficiently catalyzed the decolorization of various artificial dyes and oxidized Mn (II) to Mn (III) in the presence of $H_2O_2$. The absorption spectrum of the enzyme exhibited maxima at 407, 500, and 640 nm. The amino acid sequence of the three tryptic peptides was analyzed by ESI Q- TOF MS/MS spectrometry, and showed low similarity to those of the extracellular peroxidases of other white-rot basidiomycetes.

Reduction of Azobenzene by Purified Bovine Liver Quinone Reductase

  • Kim, Kyung-Soon;Shin, Hae-Yong
    • BMB Reports
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    • v.33 no.4
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    • pp.321-325
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    • 2000
  • Quinone reductase was purified to homogeneity from bovine liver by using ammonium sulfate fractionation, ionexchange chromatography, and gel filtration chromatography. The enzyme utilized either NADH or NADPH as the electron donor. The enzyme catalyzed the reduction of several quinones and other artificial electron acceptors. Furthermore, the enzyme catalyzed NAD(P)H-dependent reduction of azobenzene. The apparent Km for 1,4-benzoquinone and azobenzene was 1.64 mM and 0.524 mM, respectively. The reduction of azobenzene by quinone reductase was almost entirely inhibited by dicumarol or Cibacron blue 3GA, potent inhibitors of the mammalian quinone reductase. In the presence of 1.0${\mu}M$ Cibacron blue 3GA, azoreductase activity was lowered by 45%, and almost complete inhibition was seen above 2.0 ${\mu}M$ Cibacron blue 3GA.

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Evaluation of Blood compatibility at lumbrokinase immobilized polymer valves in vivo (룸브로키나제가 고정화된 폴리머 밸브의 invivo 혈액적합성 평가)

  • Park, Yong-Doo;Ryu, Eun-Sook;Kim, Jong-Won;Min, Byoung-Goo
    • Proceedings of the KOSOMBE Conference
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    • v.1998 no.11
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    • pp.227-228
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    • 1998
  • Lumbrokinase, potent fibrinolytic enzyme purified from earthworm, was immobilized onto polyurethane valves using photoreaction, photoreactive polyallyl-amino as a photoreactive linker. For evaluation of blood compatibility, lumbrokinase immobilized polymer valves were assembled into the total artificial heart (TAH). This TAH was implanted to 60kg healthy lamb for 1-3 days with the cardiac output 5 L/min. In the control lamb, the valves were untreated, in ore other, only valves on the right were treated, and in the remaining animal, only those on the left. To facilitate the thrombus formation, low doses of heparin were administered. For evaluation of the immobilized lumbrokinase, thrombus formation, proteolytic and fibrinolytic activity was measured. This data shows that lumbrokinase-treated polyurethane valves lead to decreased thrombus formation in vivo, and that their biocompatibility is therefore higher than that of untreated valves.

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Promotion of Tricholoma matsutake mycelium growth by Penicillium citreonigrum

  • Doo-Ho Choi;Jae-Gu Han;Kang-Hyo Lee;An Gi-Hong
    • Mycobiology
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    • v.51 no.5
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    • pp.354-359
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    • 2023
  • Tricholoma matsutake has been the most valuable ectomycorrhizal fungi in Asia because of its unique flavor and taste. However, due to the difficulty of artificial cultivation, the cultivation of T. matsutake has relied on natural growth in forests. To cultivate the T. matsutake artificially, microorganisms in fairy rings were introduced. In this study, we isolated 30 fungal species of microfungi from the soil of fairy rings. Among them, one single fungal strain showed a promoting effect on the growth of T. matsutake. The growth effect was confirmed by measuring the growth area of T. matsutake and enzyme activities including a-amylase, cellulase, and b-glucosidase. In comparison with control, microfungal metabolite increased the growth area of T. matsutake by 213% and the enzyme activity of T. matsutake by 110-200%. The isolated fungal strain was identified as Penicillium citreonigrum by BLAST on the NCBI database. The Discovery of this microfungal strain is expected to contribute to artificial cultivation of T. matsutake.

Cosmeceutical Properties of Fructan (Levan) Produced by Zymomonas mobilis

  • Kim, K. H.;C. S. Han;K. I. Ko;E. K. Yang;Kim, C. H.;Park, S. N.
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.700-718
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    • 2003
  • Fructan, a polysaccharide existing in plants or produced by microorganisms, is a sugar polymer of fructose with $\beta$-2,6 linkages. In this study, we investigated some cosmeceutical properties of Fructan such as moisturizing effect, cell proliferation effect, anti-inflammation effect and cell cytotoxicity. Zymomonas mobilis, a microorganism producing Fructan, was cultured in a medium containing 10% sucrose and 2% yeast extract as main components for 24 hours at 37$^{\circ}C$ and pH 7. Fructan was obtained by precipitation from the cultured medium by adding alcohol (alcohol ratio of 1:3) after removing the enzyme by centrifuging. Fructan exhibited almost same moisturizing effect as hyaluronic acid and cell proliferation effect on human fibroblast and keratinocyte as well. Moreover, on cell proliferation test on bio-artificial skin constructed by 3-dimensional(3-D) culture after inducing primary skin inflammation with 0.5% sodium lauryl sulfate (SLS), the 3-D artificial skin treated with 0.01 mg/ml, 0.05mg/ml of Fructan exhibited higher cell proliferation than the 3-D artificial skin treated with SLS only. On anti-inflammation test on 3-D artificial skin evaluated by measuring secreted quantity of interleukin-1$\alpha$ (IL-1$\alpha$) which is a pre-inflammatory mediator induced by SLS, the quantity of IL-1$\alpha$on the 3-D artificial skin treated with 0.01 mg/ml, 0.05mg/ml of Fructan was less than the one on the 3-D artificial skin treated with SLS only. As a result of these studies, Fructan has anti-inflammation effect against inflammatory reaction by a skin irritant as well as cell proliferation effect in bio-artificial skin. Fructan was also evaluated as a safe material without any toxicity in safety tests using fibroblasts and animals.

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Characterization and optimum production condition of extracellular protease from Pseudoalteromonas donghaensis HJ51 (Pseudoalteromonas donghaensis HJ51의 체외 단백질 분해효소 특성과 생산 조건)

  • Oh, Ji-Sung;Choi, Yoon-Soo;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.75-80
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    • 2015
  • Pseudoalteromonas donghaensis HJ51, isolated from the East Sea, has been reported as a novel strain to produce extracellular protease. Crude supernatant was used to determine optimal activity and optimal production conditions for the enzyme. It was found that the optimal temperature and pH of the protease were $40^{\circ}C$ and pH 7.5-10.5, respectively. The enzyme activity was kept to 88% at the pH 11. In metal requirement analysis, the enzyme exhibited the highest activity when 10 mM $Fe^{3+}$ was supplied. While supplementation of additional carbon sources used in study showed no positive effect on cell growth and enzyme activity, the addition of beef extract, tryptone, or casamino acids instead of peptone of PY-ASW containing 1% glucose increased enzyme production to 21, 7, 4%, respectively. Taken together these properties, the enzyme produced from P. donghaensis HJ51 can be applied to the industries that require protease activity under alkaline pH and low temperature.

Coal Bottom Ash Application on Park Site Soil and Its Impacts on Turfgrass Growth and Soil Quality

  • Oh, Se Jin;Kim, Yong Hyok
    • Korean Journal of Soil Science and Fertilizer
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    • v.50 no.2
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    • pp.127-134
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    • 2017
  • Bottom ash (BA) has different macro- and micronutrients such as B, Mo, Fe, Ca, and Mg, providing useful resources for plant growth and soil quality. The objective of this study was to evaluate the applicability of artificial top-soil treated with BA in park area as a vegetation base material, especially for turfgrass growth. Collected BA was mixed with peat moss and clay at the ratio of 70:10:20 (w/w). In order to evaluate the park quality and turfgrass growth in park area, the artificial soil was applied with BA along with the control or without BA. Result showed that exchangeable K and P were increased by $11.4mg\;kg^{-1}$ and $163mg\;kg^{-1}$, respectively, compared to the control soil when the artificial soil was treated with BA. Microbial population and enzyme activity (Acid-phosphatase, APA) in artificial soil having BA also increased as 2 times and 10%, respectively, compared to the control soil. Comparing turfgrass growth and yield between general soil and artificial soil, about 2 times higher plant yield (fresh weight) was observed as artificial soil was applied comparing to general soil. Furthermore, nutrient concentration in turfgrass was averaged as 0.440% for $P_2O_5$, 0.456% for CaO, 1.198% for $K_2O$ and 0.188% for MgO that all values are higher than general soil (0.323% for $P_2O_5$, 0.416% for CaO, 0.610% for $K_2O$ and 0.173% for MgO). Application of BA can be useful for vegetation base material in park site.

Enhanced Production of Succinic Acid by Metabolically Engineered Escherichia coli with Amplified Activities of Malic Enzyme and Fumarase

  • Hong, Soon-Ho;Lee, Sang-Yup
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.4
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    • pp.252-255
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    • 2004
  • A pfl ldhA double mutant Escherichia coli strain NZN 111 was used to produce succinic acid by overexpressing the E. coli malic enzyme gene (sfcA). This strain, however, produced a large amount of malic acid as well as succinic acid. After the analyses of the metabolic pathways, the fumB gene encoding the anaerobic fumarase of E. coli was co-amplified to solve the problem of malic acid accumulation. A plasmid, pTrcMLFu, was constructed, which contains an artificial operon (sfcA-fumB) under the control of the inducible trc promoter. From the batch culture of recombinant E. coli NZN 111 harboring pTrcMLFu, 7 g/L of succinic acid was produced from 20 g/L of glucose, with no accumulation of malic acid. From the metabolic flux analysis the strain was found under reducing power limiting conditions by severe reorientation of metabolic fluxes.

The effective model of the human Acetyl-CoA Carboxylase inhibition by aromatic-structure inhibitors

  • Minh, Nguyen Truong Cong;Thanh, Bui Tho;Truong, Le Xuan;Suong, Nguyen Thi Bang;Thao, Le Thi Xuan
    • Journal of IKEEE
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    • v.21 no.3
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    • pp.309-319
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    • 2017
  • The research investigates the inhibition of fatty acid biosynthesis of the human Acetyl-CoA Carboxylase enzyme by the aromatic-structure inhibitors (also known as ligands) containing variables of substituents, contributing an important role in the treatment of fatty-acid metabolic syndrome expressed by the group of cardiovascular risk factors increasing the incidence of coronary heart disease and type-2 diabetes. The effective interoperability between ligand and enzyme is characterized by a 50% concentration of enzyme inhibitor ($IC_{50}$) which was determined by experiment, and the factor of geometry structure of the ligands which are modeled by quantum mechanical methods using HyperChem 8.0.10 and Gaussian 09W softwares, combining with the calculation of quantum chemical and chemico-physical structural parameters using HyperChem 8.0.10 and Padel Descriptor 2.21 softwares. The result data are processed with the combination of classical statistical methods and modern bioinformatics methods using the statistical softwares of Department of Pharmaceutical Technology - Jadavpur University - India and R v3.3.1 software in order to accomplish a model of the quantitative structure - activity relationship between aromatic-structure ligands inhibiting fatty acid biosynthesis of the human Acetyl-CoA Carboxylase.