• Title/Summary/Keyword: K_{cat}/K_m$

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FIBREWISE INFINITE SYMMETRIC PRODUCTS AND M-CATEGORY

  • Hans, Scheerer;Manfred, Stelzer
    • Bulletin of the Korean Mathematical Society
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    • v.36 no.4
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    • pp.671-682
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    • 1999
  • Using a base-point free version of the infinite symmetric product we define a fibrewise infinite symmetric product for any fibration $E\;\longrightarrow\;B$. The construction works for any commutative ring R with unit and is denoted by $R_f(E)\;l\ongrightarrow\;B$. For any pointed space B let $G_I(B)\;\longrightarrow\;B$ be the i-th Ganea fibration. Defining $M_R-cat(B):= inf{i\midR_f(G_i(B))\longrihghtarrow\;B$ admits a section} we obtain an approximation to the Lusternik-Schnirelmann category of B which satisfies .g.a product formula. In particular, if B is a 1-connected rational space of finite rational type, then $M_Q$-cat(B) coincides with the well-known (purely algebraically defined) M-category of B which in fact is equal to cat (B) by a result of K.Hess. All the constructions more generally apply to the Ganea category of maps.

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Analysis of Antioxidant Enzyme Activity During Seedling Growth of Leymus chinensis Trin Under Salt and Dehydration Stresses (고염과 건조 스트레스 처리 조건 동안 양초 유식물체의 항산화효소 활성 분석)

  • Shim, Donghwan;Nam, Ki Jung;Kim, Yun-Hee
    • Journal of Life Science
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    • v.28 no.7
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    • pp.772-777
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    • 2018
  • To understand the adaptability of Leymus chinensis forage grass to environmental stresses, we analyzed the $H_2O_2$ scavenging activity based on several antioxidant enzymes and total phenolics content, including peroxidase (POD), ascorbate peroxidase (APX), and catalase (CAT), in shoots and roots subjected to salt and dehydration stresses during seedling growth. After NaCl or PEG treatment, plants showed reduced seedling growth under over 200 mM NaCl or 30% PEG treatment condition in shoots and roots compared with the control condition. In addition, plants showed high enzymatic activity of CAT in the shoots, whereas they exhibited high activity levels of APX and POD in the roots in both the NaCl and PEG treatment conditions. These results seem to indicate that Leymus chinensis seedlings responding to salt and dehydration stresses during initial growth is associated with enhanced activity of $H_2O_2$ scavenging antioxidant enzymes in the shoots or roots. The plants also showed high levels of total phenolics under NaCl treatment, with a high concentration in both the shoots and roots. Our results showed that the induced activity patterns of APX in the roots and CAT in the shoots indicate that low $H_2O_2$ levels were mainly maintained through tissue-specific redox homeostasis involving enzymes such as APX and CAT during salt and dehydration stresses. This study highlights the importance of antioxidant enzymes in the establishment of Leymus chinensis seedlings under high salinity conditions, such as typical desertification.

Establishment of Cryopreservation of Leopard Cat Semen Collected by Electro-ejaculation Method

  • Ha, A-Na;Jo, A-Ra;Kim, Yu-Gon;Yoon, Jin-Ho;Bang, Jae-Il;Deb, Gautam K.;Fakruzzaman, M.;Lim, Yang-Mook;Yong, Hwan-Yul;Kong, Il-Keun
    • Journal of Embryo Transfer
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    • v.26 no.4
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    • pp.245-250
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    • 2011
  • The aim of this study was to evaluate the post-thawed characteristics of leopard cat semen. In this experiment, semen was collected from two leopard cats (A and B) at wild animal center in Seoul Grand Park in Korea. After collection, the sperms were washed with D-PBS and diluted by the freezing medium (Irvine science, USA) and stored in liquid nitrogen. The post-thawed concentration was $357{\times}10^6sperms/ml$ for A and $97{\times}10^6sperms/ml$ for B. The viability of post-thawed sperm from A and B individual was 24.0% and 19.0%, respectively. Pre-freezing motility of A and B individual semen was 68.54% and 56.65. Leopard cat A had more normal sperm than that of B (69.5% vs. 54.5%). Acrosome integrity analysis detected live (14.5% vs. 9.0%), damage (39.0% vs. 44.0%) and dead (46.0% vs. 47.0%) in leopard cat A and B, respectively. The present results concluded that leopard cat semen can be collected successfully by electro-ejaculation method and cryopreserved successfullyfor future use in different assisted reproductive technologies. The cryopreservation protocol needs to be modified for increasing post-thawed viability of leopard cat spermatozoa.

Heteroexpression and Functional Characterization of Glucose 6-Phosphate Dehydrogenase from Industrial Aspergillus oryzae

  • Guo, Hongwei;Han, Jinyao;Wu, Jingjing;Chen, Hongwen
    • Journal of Microbiology and Biotechnology
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    • v.29 no.4
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    • pp.577-586
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    • 2019
  • The engineered Aspergillus oryzae has a high NADPH demand for xylose utilization and overproduction of target metabolites. Glucose-6-phosphate dehydrogenase (G6PDH, E.C. 1.1.1.49) is one of two key enzymes in the oxidative part of the pentose phosphate pathway, and is also the main enzyme involved in NADPH regeneration. The open reading frame and cDNA of the putative A. oryzae G6PDH (AoG6PDH) were obtained, followed by heterogeneous expression in Escherichia coli and purification as a his6-tagged protein. The purified protein was characterized to be in possession of G6PDH activity with a molecular mass of 118.0 kDa. The enzyme displayed maximal activity at pH 7.5 and the optimal temperature was $50^{\circ}C$. This enzyme also had a half-life of 33.3 min at $40^{\circ}C$. Kinetics assay showed that AoG6PDH was strictly dependent on $NADP^+$ ($K_m=6.3{\mu}M$, $k_{cat}=1000.0s^{-1}$, $k_{cat}/K_m=158.7s^{-1}{\cdot}{\mu}M^{-1}$) as cofactor. The $K_m$ and $k_{cat}/K_m$ values of glucose-6-phosphate were $109.7s^{-1}{\cdot}{\mu}M^{-1}$ and $9.1s^{-1}{\cdot}{\mu}M^{-1}$ respectively. Initial velocity and product inhibition analyses indicated the catalytic reaction followed a two-substrate, steady-state, ordered BiBi mechanism, where $NADP^+$ was the first substrate bound to the enzyme and NADPH was the second product released from the catalytic complex. The established kinetic model could be applied in further regulation of the pentose phosphate pathway and NADPH regeneration of A. oryzae to improve its xylose utilization and yields of valued metabolites.

Expression of Catalase (CAT) and Ascorbate Peroxidase (APX) in MuSI Transgenic Tobacco under Cadmium Stress

  • Kim, Kye-Hoon;Kim, Young-Nam;Lim, Ga-Hee;Lee, Mi-Na;Jung, Yoon-Hwa
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.1
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    • pp.53-57
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    • 2011
  • The MuSI is known as a multiple stress resistant gene with several lines. A previous study using RT-PCR showed that the expression of MuSI gene in tobacco plant induced its tolerance to Cd stress. This study was conducted to examine the enhanced Cd tolerance of the MuSI transgenic tobacco plant through germination test and to understand the role of the involved antioxidant enzymes for the exhibited tolerance. Germination rate of MuSI transgenic tobacco was more than 10% higher than that of wild-type tobacco, and seedlings of MuSI transgenic tobacco grew up to 1.6 times larger and greener than seedlings of wild-type tobacco at 200 and 300 ${\mu}M$ Cd. From the third to the fifth day, CAT activities at 100 and 200 ${\mu}M$ Cd and APX activities at 100, 200 and 300 ${\mu}M$ Cd of MuSI transgenic tobacco were up to two times higher than those of wild-type tobacco. MuSI gene is shown to enhance the activities of antioxidant enzymes resulting in higher tolerance to oxidative stress compared with the control plant.

Kinetic Characterization of an Iron-sulfur Containing Enzyme, L-serine Dehydratase from Mycobacterium tuberculosis H37Rv (Mycobacterium tuberculosis H37Rv로부터 유래된 철-황 함유 효소인 L-세린 탈수화효소의 동력학적 특성)

  • Han, Yu Jeong;Lee, Ki Seog
    • Journal of Life Science
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    • v.28 no.3
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    • pp.351-356
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    • 2018
  • L-Serine dehydratase (LSD) is an iron-sulfur containing enzyme that catalyzes the conversion of L-serine to pyruvate and ammonia. Among the bacterial amino acid dehydratases, it appears that only the L-serine specific enzymes utilize an iron-sulfur cluster at their catalytic site. Moreover, bacterial LSDs are classified into four types based on structural characteristics and domain arrangement. To date, only the LSD enzymes from a few bacterial strains have been studied, but more detailed investigations are required to understand the catalytic mechanism of various bacterial LSDs. In this study, LSD type II from Mycobacterium tuberculosis (MtLSD) H37Rv was expressed and purified to elucidate the biochemical and catalytic properties using the enzyme kinetic method. The L-serine saturation curve of MtLSD exhibited a typically sigmoid character, indicating an allosteric cooperativity. The values of $K_m$ and $k_{cat}$ were estimated to be $59.35{\pm}1.23mM$ and $18.12{\pm}0.20s^{-1}$, respectively. Moreover, the plot of initial velocity versus D-serine concentration at fixed L-serine concentrations showed a non-linear hyperbola decay shape and exhibited a competitive inhibition for D-serine with an apparent $K_i$ value of $30.46{\pm}5.93mM$ and with no change in the $k_{cat}$ value. These results provide insightful biochemical information regarding the catalytic properties and the substrate specificity of MtLSD.

Biomimetic Balancing Mechanism for Walking Robot (생체모사를 통한 보행로봇의 균형감에 관한 연구)

  • Kim, Jong Jin;Chung, Seong Youb
    • Journal of Institute of Convergence Technology
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    • v.4 no.2
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    • pp.55-59
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    • 2014
  • A cat is able to quickly recover balance from unstable posture. To observe the balance recovery procedure of the cat, an impulse is applied to the cat while walking on a narrow bridge. We find that it rotates its tail toward the falling direction. In our previous research, the balance recovery procedure is analyzed based on the law of the angular momentum conservation and then a key equation is derived to maintain the balance. However, it did not consider the gravity, so the performance is not good. In this paper, a new dynamic model is proposed using the Lagrangian mechanics. In the method, the gravity is included in the potential energy. Through the proposed dynamic model, controlling the balance of a walking robot is possible.

A Numerical Study on Clear-Air Turbulence Events Occurred over South Korea (한국에서 발생한 청천난류 사례들에 대한 수치연구)

  • Min, Jae-Sik;Kim, Jung-Hoon;Chun, Hye-Yeong
    • Atmosphere
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    • v.22 no.3
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    • pp.321-330
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    • 2012
  • Generation mechanisms of the three moderate-or-greater (MOG)-level clear-air turbulence (CAT) encounters over South Korea are investigated using the Weather Research and Forecasting (WRF) model. The cases are selected among the MOG-level CAT events occurred in Korea during 2002-2008 that are categorized into three different generation mechanisms (upper-level front and jet stream, anticyclonic flow, and mountain waves) in the previous study by Min et al. For the case at 0127 UTC 18 Jun 2003, strong vertical wind shear (0.025 $s^{-1}$) generates shearing instabilities below the enhanced upper-level jet core of the maximum wind speed exceeding 50 m $s^{-1}$, and it induces turbulence near the observed CAT event over mid Korea. For the case at 2330 UTC 22 Nov 2006, areas of the inertia instability represented by the negative absolute vorticity are formed in the anticyclonically sheared side of the jet stream, and turbulence is activated near the observed CAT event over southwest of Korea. For the case at 0450 UTC 16 Feb 2003, vertically propagating mountain waves locally trigger shearing instability (Ri < 0.25) near the area where the background Richardson number is sufficiently small (0.25 < Ri < 1), and it induces turbulence near the observed CAT over the Eastern mountainous region of South Korea.

In vivo Physiological Activity of Mentha viridis L. and Mentha piperita L. (박하의 in vivo 생리활성)

  • Lee, Seung-Eun;Han, Hee-Sun;Jang, In-Bok;Kim, Geum-Soog;Seong, Nak-Sul
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.6
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    • pp.261-267
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    • 2005
  • Alcohol metabolizing and antioxidant activity of Mentha species were investigated in rat liver. Fifty six Sprague Dawley rats were randomly divided into seven groups such as normal (ethanol excluded), negative control (40% ethanol (10 g/kg of body weight/day) fed), positive control (1 g Silymarin/kg of body weight/day with ethanol fed), two Mentha viridis extracts (0.2 g & 1 g M. viridis methanol ext./kg of body weight/day with ethanol fed) and two M piperita extracts (0.2 g & 1 g M. piperita methanol ext./kg of body weight/day with ethanol fed) groups. After 2 weeks, rats were sacrificed under ether. The activities of alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH), catalase (CAT), manganese superoxide dismutase (Mn-SOD), glutathione peroxidase (GAH-px) and the content ofthiobarbituric acid reactive substance (TBARS) in the rat livers and the activity of glutamate pyruvate transferase (GPT) in serum were evaluated. From the analyses, 1 g M. viridis and 0.2 g M. piperita administrated groups showed higher ADH and ALDH activity than the other groups. Groups fed with 0.2 g and 1 g M. viridis ext. and 0.2 g M. piperita ext. showed higher CAT activity than the other groups. All the Mentha extract fed groups exhibited more effective in recovering Mn-SOD, GSH-px and GPT acitivities to a similar degree of normal group. TBARS contents of two M. viridis ext. fed group and 0.2 g M. piperita ext. fed group were higher than those of the other groups. M. viridis extract fed groups showed more effective in CAT and Mn-SOD activities than M. piperita extract groups at p < 0.05. Finally, it is concluded that both Mentha species have alcohol metabolizing and antioxidant activity and M viridis is more effective than M. piperita.

Determination of the Kinetic Properties of Platy cod in D for the Inhibition of Pancreatic Lipase Using a 1, 2-Diglyceride- Based Colorimetric Assay

  • Zhao, Hai-Un;Kim, Yeong-Shik
    • Archives of Pharmacal Research
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    • v.27 no.9
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    • pp.968-972
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    • 2004
  • A 1, 2-diglyceride-based multi-step colorimetric assay to measure the pancreatic lipase activ-ity was applied for the determination of the kinetic profiles of the lipase inhibition with a slight modification and the validity verification. With this assay method, our study revealed that platy-codin D, one of major constituents of Platycodi Radix, inhibits the pancreatic lipase activity in a competitive type, with the value of $K_1$ being 0.18${\pm}$0.02 mM. In addition, PO has affected the val-ues of $K_{m}$, app/ and $K_{cat}$/$K_{m}$ in a dose-dependent manner. The results shed a meaningful light on how PO mediates lipid metabolism in the intestinal tracts. On the other hand, since the revised assay is sensitive, rapid, and does not affect the accuracy to the kinetic properties, it is applica-ble not only to evaluation of the kinetic properties of the pancreatic lipase, but also to high-throughput screening of pancreatic lipase activity.