• Title/Summary/Keyword: K_{cat}/K_m$

Search Result 395, Processing Time 0.036 seconds

Effects of Structural Difference of Ionic Liquids on the Catalysis of Horseradish Peroxidase

  • Hong, Eun-Sik;Park, Jung-Hee;Yoo, Ik-Keun;Ryu, Keun-Garp
    • Journal of Microbiology and Biotechnology
    • /
    • v.19 no.7
    • /
    • pp.713-717
    • /
    • 2009
  • The dependence of the catalytic properties of horseradish peroxidase on the structural changes of ionic liquids was investigated with two water-miscible ionic liquids, N-butyl-3methypyridinium tetraftuoroborate ([$BMP_y$][$BF_4$]) and 1-butyl-3-methylimidazolium methylsulfate ([BMIM][$MeSO_4$]), each of which shares an anion ($BF_4^-$) or a cation ($BMIM^+$) with 1-butyl-3-methylimidazolium tetraftuoroborate ([BMIM][$BF_4$]), respectively. The oxidation of guaiacol (2-methoxyphenol) with $H_2O_2$was used as a model reaction. In order to minimize the effect of solution viscosity on the kinetic constants of the enzymatic catalysis, the enzymatic reactions for the kinetic study were performed in water-ionic liquid mixtures containing 25% (v/v) ionic liquid at maximum. Similarly to the previously reported results for [BMIM][$BF_4$], as the concentration of [$BMP_y$][$BF_4$] increased, the $K_m$value increased with a decrease in the $k_{cat}$value: the $K_m$value increased markedly from 2.8 mM in 100% water to 12.6 mM in 25% (v/v) ionic liquid, indicating that ionic liquid significantly weakens the binding affinity of guaiacol to the enzyme. On the contrary, [BMIM][$MeSO_4$] decreased the Km value to 1.4 mM in 25% (v/v) ionic liquid. [BMIM][$MeSO_4$] also decreased $k_{cat}$more than 3-folds [from 13.8 $s^{-1}$in 100% water to 4.1 $s^{-1}$in 25% (v/v) ionic liquid]. These results indicate that the ionic liquids interact with the enzyme at the molecular level as well as at a macroscopic thermodynamic scale. Specifically, the anionic component of the ionic liquids influenced the catalysis of horseradish peroxidase in different ways.

Community-acquired Extended-spectrum and Plasmid-mediated ampC Beta-lactamase-producing Multidrug-resistant Enterobacter cloacae Septicaemia in a Cat with Euthyroid Sick Syndrome (정상 갑상샘 질환 증후군 고양이의 지역사회획득 광범위 및 플라스미드 유래 ampC beta-lactamase 양성 다약제내성 Enterobacter cloacae 패혈증)

  • Han, Jae-Ik;Na, Ki-Jeong
    • Journal of Veterinary Clinics
    • /
    • v.32 no.2
    • /
    • pp.191-195
    • /
    • 2015
  • A 7-year-old castrated male Korean Shorthair cat was referred with lethargy and anorexia. Laboratory examination revealed moderate degenerative changes of peripheral neutrophils on blood smear examination and decreased levels of free and total thyroxine ($T_4$) as well as bacterial growth on blood culture. Molecular analyses of the 16S ribosomal RNA gene and heat shock protein 60 gene confirmed the bacterium as Enterobacter cloacae. A minimal inhibitory concentration test showed multidrug resistance of the bacterium against 16 antibiotics. Polymerase chain reaction (PCR) and subsequent sequencing specifically for $bla_{TEM}$, $bla_{SHV}$, $bla_{CTX-M}$, and plasmid-mediated ampC genes revealed positive results to $bla_{TEM-1}$, $bla_{CTX-M-15}$, and plasmid-mediated $bla_{ACT-1}$ genes, indicating that the isolated bacterium contains plasmids containing genes encoding extended-spectrum beta-lactamase and plasmid-mediated ampC beta-lactamase. After 1 month of treatment with antibiotics and levothyroxine, the cat's condition improved; both the thyroid function test and the blood culture showed no abnormalities. This is the first report of community-acquired multidrug-resistant E. cloacae-induced euthyroid sick syndrome in a cat. By the prompt diagnostic procedures and properly selected antibiotic therapy, the cat was recovered from the multidrug-resistant bacterium-induced septicaemia.

Reproductivity of Cloned Male Cat

  • Choi, E.G.;Lee, H.S.;Yin, X.J.;Ho, J.H.;Cho, E.M.;Kang, S.H.;Bae, I.H.;Cho, S.K.;Kong, I.K.
    • Proceedings of the Korean Society of Developmental Biology Conference
    • /
    • 2005.10a
    • /
    • pp.129-129
    • /
    • 2005
  • PDF

Reproductivity of Cloned Male Cat

  • Choi, E.G.;Lee, H.S.;Yin, X.J.;Oh, J.H.;Cho, E.M.;Kang, S.H.;Bae, I.H.;Cho, S.K.;Kong, I.K.
    • Proceedings of the Korean Society of Embryo Transfer Conference
    • /
    • 2005.10a
    • /
    • pp.129-129
    • /
    • 2005
  • PDF

Biochemical Analysis on the Parallel Pathways of Methionine Biosynthesis in Corynebacterium glutamicum

  • Hwang, Byung-Joon;Park, Soo-Dong;Kim, Youn-Hee;Kim, Pil;Lee, Heung-Shick
    • Journal of Microbiology and Biotechnology
    • /
    • v.17 no.6
    • /
    • pp.1010-1017
    • /
    • 2007
  • Two alternative pathways for methionine biosynthesis are known in Corynebacterium glutamicum: one involving transsulfuration (mediated by metB and metC) and the other involving direct sulfhydrylation (mediated by metY). In this study, MetB (cystathionine ${\gamma}-synthase$) and MetY (O-acetylhomoserine sulfhydrylase) from C. glutamicum were purified to homogeneity and the biochemical parameters were compared to assess the functional and evolutionary importance of each pathway. The molecular masses of the native MetB and MetY proteins were measured to be approximately 170 and 280 kDa, respectively, showing that MetB was a homotetramer of 40-kDa subunits and MetY was a homohexamer of 45-kDa subunits. The $K_m$ values for the O-acetylhomoserine catalysis effected by MetB and MetY were 3.9 and 6.4 mM, and the maximum catalysis rates were $7.4\;(k_{cat}=21\;S^{-1})\;and\;6.0\;(k_{cat}=28\;S^{-1})\;{\mu}mol\;mg^{-1}\;min^{-1}$, respectively. This suggests that both MetB and MetY can be comparably active in vivo. Nevertheless, the $K_m$ value for sulfide ions by MetY was 8.6mM, which was too high, considering the physiological condition. Moreover, MetB was active at a broad range of temperatures $(30\;and\;65^{\circ}C)$ and pH (6.5 and 10.0), as compared with MetY, which was active in a range from 30 to $45^{\circ}C$ and at pH values from 7.0 to 8.5. In addition, MetY was inhibited by methionine, but MetB was not. These biochemical data may provide insight on the role of the parallel pathways of methionine biosynthesis in C. glutamicum with regard to cell physiology and evolution.

Inabenfide-Induced Alleviation of Salt Stress in Rice as Linked to Changes in Salicylic Acid Content and Catalase Activity

  • Sawada, Hiroko;Kim, Dea-Wook;Kobayashi, Katsuichiro;Shim, Ie-Sung
    • Journal of Crop Science and Biotechnology
    • /
    • v.10 no.1
    • /
    • pp.39-44
    • /
    • 2007
  • The effect of inabenfide was investigated in rice seedlings subjected to salt stress in relation to changes in chlorophyll fluorescence(${\Delta}F/Fm'$), lipid peroxidation, salicylic acid(SA) content, and catalase(CAT) activity. A reduction in shoot growth of rice seedlings by 120 mM NaCl treatment was significantly alleviated by pretreatment with 30 ${\mu}M$ inabenfide. Sodium ion content was not affected by pretreatment with inabenfide, suggesting that alleviation was not due to a reduction in sodium ion uptake by the rice seedlings. At three days after NaCl treatment, the rice seedlings pretreated with inabenfide showed a higher ${\Delta}F/Fm'$(30%) and lower lipid peroxidation(28%) compared with the rice seedlings treated with NaCl alone. After NaCl treatment, CAT activity in the third leaf of rice seedlings decreased significantly but alleviated by pretreatment with inabenfide. Furthermore, pretreatment with inabenfide also reduced the level of SA which accumulated drastically in the third leaf of rice seedlings within a day after exposure to salt stress. These results suggest that inabenfide prevents SA accumulation in rice seedlings under salt stress which eventually induces the alleviation of salt stress damage.

  • PDF

The Airflow Obstruction and Subjective Health Status Among Stable Chronic Obstructive Pulmonary Disease Patients Residing in the Community (안정된 만성폐쇄성폐질환 환자의 기류제한 정도와 주관적 건강상태)

  • Song, Hee-Young
    • Journal of Korean Biological Nursing Science
    • /
    • v.19 no.1
    • /
    • pp.38-47
    • /
    • 2017
  • Purpose: This study was undertaken to examine the relationship between airflow obstruction and subjective health status reported by stable patients with chronic obstructive pulmonary disease (COPD) residing in the community. Methods: A cross-sectional descriptive study was conducted with 78 stable COPD patients aged 69.7 years old on average and selected by a convenient sampling from an outpatient department of pulmonology in tertiary hospitals. They completed a constructed questionnaire including general characteristics, smoking history, dyspnea by modified medical research council (mMRC) scale, and health status by COPD assessment test (CAT). Anthropometric measurements were performed for body mass index (BMI) and pulse oxymetry for $O_2$ saturation (Sat $O_2$). Medical records were reviewed to obtain disease-related characteristics including duration of the disease, cardiovascular comorbidity, and forced expiratory volume in 1 second ($FEV_1$). Data were analyzed using PASW statistics 20.0. Results: Mean $FEV_1%$ and CAT scores were 55.11% and 17.73, respectively. Those in the lower stage of mMRC showed significantly higher $FEV_1$ and lower CAT. $FEV_1$ and CAT showed significant negative correlations; age and BMI with $FEV_1$, and Sat $O_2$ with CAT. Conclusion: The findings suggest that the less airway obstruction was, the better health status was, and provide the support for using subjective measures in clinical practices for COPD patients.

Protective effect of the methanol extract of Polyopes lancifolia (Harvey) kawaguchi et wang against ionizing radiation-induced mouse gastrointestinal injury

  • Jeong, Jinwoo;Yang, Wonjun;Ahn, Meejung;Kim, Ki Cheon;Hyun, Jin Won;Kim, Sung-Ho;Moon, Changjong;Shin, Taekyun
    • Korean Journal of Veterinary Research
    • /
    • v.51 no.3
    • /
    • pp.177-183
    • /
    • 2011
  • The radioprotective efficacy of a methanol extract of the red algae Polyopes lancifolia (Harvey) kawaguchi et wang (mPL) was evaluated in mice subjected to total-body gamma irradiation. mPL protection against radiation-induced oxidative stress was examined by histological evaluation of intestinal crypt-cell survival and liver activities of the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). mPL (100 mg/kg body weight) administered intraperitoneally at 24 h and 1 h prior to irradiation protected jejunal crypt cells from radiation-induced apoptosis (p < 0.01). The pretreatment of mPL attenuated a radiation-induced decrease in villous height (p < 0.05), and improved jejunal crypt survival (p < 0.05). The dose reduction factor was 1.14 at 3.5 days after irradiation. Treatment with mPL prior to irradiation resulted in significantly higher (p < 0.01) levels of SOD and CAT activities, compared to those levels of irradiated control mice with vehicle treatment. These results suggest that mPL is a useful radioprotective agent capable of defending intestinal progenitor cells against total-body irradiation, at least in part through mPL antioxidative activity.

Echinostoma macrorchis in Lao PDR: Metacercariae in Cipangopaludina Snails and Adults from Experimentally Infected Animals

  • Sohn, Woon-Mok;Chai, Jong-Yil;Na, Byoung-Kuk;Yong, Tai-Soon;Eom, Keeseon S.;Park, Hansol;Min, Duk-Young;Rim, Han-Jong
    • Parasites, Hosts and Diseases
    • /
    • v.51 no.2
    • /
    • pp.191-196
    • /
    • 2013
  • The echinostome metacercariae encysted in Cipangopaludina sp. snails that were purchased from a market in Vientiane Municipality, Lao PDR, were identified as Echinostoma macrorchis (Digenea: Echinostomatidae) through recovery of adult flukes after experimental infection to rats and a cat. The metacercariae were round, $113-128(121){\times}113-125(120){\mu}m$, having a thick cyst wall, a head collar armed with collar spines, and excretory granules. The adult flukes recovered from the rats and cat at day 14 and 30 post-infection, respectively, were elongated, ventrally curved, and $3.9-6.3{\times}0.7-1.1mm$ in size. The head collar was distinct, bearing 43-45 collar spines with 5 angle spines on each side. Two testes were large (as the name implies), tandem, and slightly constricted at the middle, with irregular margins. Eggs were operculated, ovoid to elliptical, and $88-95{\times}56-60{\mu}m$. In scanning electron microscopy, the head collar was prominent, with 43-45 collar spines. Scale-like tegumental spines were densely distributed on the ventral surface between the oral and ventral suckers. Sensory papillae were distributed mainly on the tegument around the 2 suckers. It is confirmed that E. macrorchis is distributed in Lao PDR using Cipangopaludina sp. snails as the second intermediate host.

Hydroxylation of Compactin (ML-236B) by CYP105D7 (SAV_7469) from Streptomyces avermitilis

  • Yao, Qiuping;Ma, Li;Liu, Ling;Ikeda, Haruo;Fushinobu, Shinya;Li, Shengying;Xu, Lian-Hua
    • Journal of Microbiology and Biotechnology
    • /
    • v.27 no.5
    • /
    • pp.956-964
    • /
    • 2017
  • Compactin and pravastatin are competitive cholesterol biosynthesis inhibitors of 3-hydroxy-3-methylglutaryl-CoA reductase and belong to the statin drugs; however, the latter shows superior pharmacokinetic characteristics. Previously, we reported that the bacterial P450, CYP105D7, from Streptomyces avermitilis can catalyze the hydroxylation of 1-deoxypentalenic acid, diclofenac, and naringenin. Here, we demonstrate that CYP105D7 could also catalyze compactin hydroxylation in vitro. In the presence of both bacterial and cyanobacterial redox partner systems with an NADPH regeneration system, the reaction produced two hydroxylated products, including pravastatin (hydroxylated at the C6 position). The steady-state kinetic parameters were measured using the redox partners of putidaredoxin and its reductase. The $k_m$ and $k_{cat}$ values for compactin were $39.1{\pm}8.8{\mu}M$ and $1.12{\pm}0.09min^{-1}$, respectively. The $k_{cat}/K_m$ value for compactin ($0.029min^{-1}{\cdot}{\mu}M^{-1}$) was lower than that for diclofenac ($0.114min^{-1}{\cdot}{\mu}M^{-1}$). Spectroscopic analysis showed that CYP105D7 binds to compactin with a $K_d$ value of $17.5{\pm}3.6{\mu}M$. Molecular docking analysis was performed to build a possible binding model of compactin. Comparisons of different substrates with CYP105D7 were conclusively illustrated for the first time.