• Title/Summary/Keyword: inactivation.

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Conformational Lock and Dissociative Thermal Inactivation of Lentil Seedling Amine Oxidase

  • Moosavi-Nejad, S. Zahra;Moosavi-Movahedi, Ali-Akbar;Rezaei-Tavirani, Mostafa;Floris, Giovanni;Medda, Rosaria
    • BMB Reports
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    • v.36 no.2
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    • pp.167-172
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    • 2003
  • The kinetics of thermal inactivation of copper-containing amine oxidase from lentil seedlings were studied in a 100 mM potassium phosphate buffer, pH 7, using putrescine as the substrate. The temperature range was between $47-60^{\circ}C$. The thermal inactivation curves were not linear at 52 and $57^{\circ}C$; three linear phases were shown. The first phase gave some information about the number of dimeric forms of the enzyme that were induced by the higher temperatures using the "conformational lock" pertaining theory to oligomeric enzyme. The "conformational lock" caused two additional dimeric forms of the enzyme when the temperature increased to $57^{\circ}C$. The second and third phases were interpreted according to a dissociative thermal inactivation model. These phases showed that lentil amine oxidase was reversibly-dissociated before the irreversible thermal inactivation. Although lentil amine oxidase is not a thermostable enzyme, its dimeric structure can form "conformational lock," conferring a structural tolerance to the enzyme against heat stress.

Chemical Modification of Serratia marcescens Acetolactate Synthase with Cys, Trp, and Arg Modifying Reagents

  • Choi, Ho-Il;Kim, Soung-Soo
    • BMB Reports
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    • v.28 no.1
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    • pp.40-45
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    • 1995
  • Acetolactate synthase purified from Serratia marcescens ATCC 25419 was rapidly inactivated by the thiol specific reagent p-chloromercuribenzoate (PCMB), the tryptophan specific reagent N-bromosuccinimide (NBS), and the arginine modifying reagent phenylglyoxal (PGO). Inactivation by PCMB was prevented by both ${\alpha}$-ketobutyrate and pyruvate, and the second order rate constant for the inactivation was $2480\;M^{-1}{\cdot}min^{-1}$. The reaction order with respect to PCMB was 0.94. The inactivation of the enzyme by NBS was also substantially reduced by both ${\alpha}$-ketobutyrate and pyruvate. The second order rate constant for inactivation by NBS was $15,000\;M^{-1}{\cdot}min^{-1}$, and the reaction order was 2.0. On the other hand, inactivation by PGO was partially prevented by ${\alpha}$-ketobutyrate, but not by pyruvate. The second order rate constant for the inactivation was $1480\;M^{-1}{\cdot}min^{-1}$ and the order of reaction with respect to PGO was 0.75. These results suggest that essential cysteine, tryptophan and arginine are located at or near the substrate binding site.

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Study on the inactivation of intracellular enzyme mlecules by X-ray irradiation (X-선조사(線照射)에 의한 세포내효소분자(細胞內酵素分子)의 불활성화(不活性化)에 관한 연구(硏究))

  • Lee, Sang-Bok
    • Journal of Radiation Protection and Research
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    • v.2 no.1
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    • pp.31-37
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    • 1977
  • Inactivation of the glutamic acid dehydrogenase and glucose-6-phosphate dehydrogenase enzyme-molecules in the Ehrlich ascites tumor cells of the mouse were studied. The above mentioned intracellular enzymemolecules were irradiated by the X-ray radiation under the condition of 65 kV, I Amp. under the atmosphere of nitrogen gases and by $4^{\circ}C$. Thereby, irradiation doses were 580 KR/min($error:{\pm}3%$). After irradiation, the cell homogentes were prepared through liquid air techniquese. There after, the activities of the enzymes were measured with photometric method given by O. Warburg and W. Christian. The dose effect curves of the activities of the two enzymes by the X-ray irradiation showed both exponential and the inactivation doses were $6,5.10^{0}\;and\;5,0.10^{6}$ R respectively. These results showed one side that the inactivation process of the intracelluar enzymemolecules was one hit reaction after target theory, and the other side that this inactivation process could not be the primary causes of the death through X-ray irradiation of the vertebrate animals, because of the high resistance of the intracellular protein molecules against X-ray irradiation. The one hit reaction by the inactivation process of the irradiated intracellular enzymemolecules was discussed.

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Chemical Modification of Glycolate Oxidase from Spinach by Diethyl Pyrocarbonate. Evidence of Essential Histidine for Enzyme Activity$^\dag$

  • Lee, Kun-Kook;Kim, Hong-Sun;Choi, Jung-Do
    • Bulletin of the Korean Chemical Society
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    • v.8 no.4
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    • pp.280-285
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    • 1987
  • FMN-dependent glycolate oxidase from spinach is inactivated by diethyl pyrocarbonate at pH 7.0. Inactivation of both apo- and holoenzyme by diethyl pyrocarbonate follows pseudo-first-order kinetics and first order with respect to the reagent. A series of difference spectra of inactivated and native enzymes show a single peak at 240 nm, indicating the modification of histidyl residues. No decrease in absorbance at around 280 nm due to formation of O-carbethoxytyrosine is observed. The rate of inactivation is dependent on pH, and the data for pH dependent rates implicate the involvement of a group with a pKa of 6.9. The activity lost by treatment with diethyl pyrocarbonate could be almost fully restored by incubation with 0.75M hydroxylamine. The reactivation by hydroxylamine and the pH dependence of inactivation are also consistent with that the inactivation is due to modification of histidyl residues. Although coenzyme FMN is without protective effect, the substrate glycolate, the product glyoxylate, and two competitive inhibitors, oxalate and oxalacetate, provide marked protection against the inactivation of the holoenzyme. These results suggest that the inactivation of the oxidase by diethyl pyrocarbonate occurs by modification of essential histidyl residue(s) at the active site.

Inactivation Effect of Cryptosporidium by Ozone and UV (Ozone과 UV를 이용한 Cryptosporidium의 불활성화 효과)

  • Kim, Yun-Hee;Lee, Chul-Hee;Lee, Shun-Hwa
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.1
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    • pp.31-39
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    • 2007
  • The objective of this study was to investigate the inactivation characteristics of Cryptosporidium oocysts by ozone and UV and to suggest the better, disinfection method. The inactivation CT value of 1 log(90%) and 2 log(99%) in of one disinfection, which is an index of disinfection for inactivation effect by ozone, were respectively 5.77 $mg{\cdot}min/L$ and 21.30 $mg{\cdot}min/L$. The inactivation in UV disinfection was not affected by pHs(5, 7 and 9), low turbidity(5 and below NTU) and UV intensity(0.2 and 0.6 $mWs/cm^2$) but obviously decreased at high turbidity over 20 NTU. Therefore UV disinfection capacity can be obtained when a good turbidity removal in drinking water treatment process is achieved. And if oocysts is exposed by high UV over 0.6 mWs/cm2 during enough time, the better inactivation effect will be obtained.

Prediction of Cryptosporidium parvum Inactivation in Advanced Ozone Drinking Water Treatment with Lab Scale Experiments (실험실 규모 크립토스포리디움의 불활성화 실험을 통한 오존 고도정수처리 정수장에서 소독 효과 예측)

  • Cho, Min;Chung, Hyenmi;Kim, Reeho;Shon, Jinsik;Park, Sangjung;Yoon, Jeyong
    • Journal of Korean Society on Water Environment
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    • v.21 no.1
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    • pp.7-13
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    • 2005
  • With the appearance of pathogenic microorganisms, which were resistant to free chlorine, the significant attention to the necessity of powerful alternative disinfection methods such as ozone, chlorine dioxide, LTV irradiation to inactivating pathogens has been increased in water treatment. Among these alternative disinfection methods, ozone is well known as strong biocidal method and the usage of ozone is also increasing in Korea. However, in Korea, there has been no report on the quantitative study of Cryptosporidium parvum with ozone and its evaluation in advanced drinking water treatments. This study reports on the methodology for predicting the ozone inactivation of Cryptosporidium parvum by ozone disinfection in advanced drinking water treatment. The method is based on the fact that a specific inactivation level of microorganisms is achieved at a unique value of ozone exposures, independent of ozone dose and type of water, and quantitatively described by a delayed Chick-Watson model. The required values ${\bar{C}}T$ for 2 log inactivation of Cryptosporidium parvum was $6.0mg/L{\cdot}min$ and $15.5mg/L{\cdot}min$ at $20^{\circ}C$ and $5^{\circ}C$, respectively. From this obtained Cryptosporidium parvum inactivation curves and calculated ${\bar{C}}T$ values of advanced drinking water treatment water in Korea with FIA (Flow injection alaysis), we can predict that water treatment plant can achieve a 1.1~1.8 log inactivation and 0~0.4 log inactivation at $20^{\circ}C$ and $5^{\circ}C$, respectively. This methodology will be useful for drinking water treatment plants which intend to evaluate the disinfection efficiencies of their ozonation process without full scale test and direct experiments with Cryptosporidium parvum.

A Study of Sterilization Effect of Long-wavelength UVA-LED Irradiation on Bacteria Causing Eye Diseases (장파장의 자외선 LED 광원을 이용한 안질환 세균의 살균효과)

  • Lee, Cheol-Woo;Jeong, Kyeong-In;Hwang, Kwang-Ha;Lee, Seok-Ju;Yoo, Geun-Chang
    • Journal of Korean Ophthalmic Optics Society
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    • v.17 no.1
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    • pp.99-105
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    • 2012
  • Purpose: The purpose of this study is to demonstrate inactivation effect of UVA-LED ultraviolet radiation upon Pseudomonas aeruginosa and Staphylococcus aureus which are the major bacteria causing eye diseases. Methods: The small sterilization device was made using UVA-LED of 400 nm. After Pseudomonas aeruginosa was diluted to $10^{-7}$ and Staphylococcus aureus to $10^{-5}$ and diluted solutions were put onto each liquid medium. They were irradiated by 400 nm of UVA for different amount of time; 15 min, 30 min, 60 min, 120 min, 240 min, 360 min and 480 min each. Results: The data from sterilization test was solved to regression line equation and the target log inactivation was obtained. The 3 log inactivation UV irradiation value of Pseudomonas aeruginosa was 54,847 UV dose ($mJ/cm^2$) and irradiation time was 135.42 min while the 3 log inactivation of Staphylococcus aureus was 39,066 UV dose ($mJ/cm^2$) and irradiation time was 98.72 min. Conclusions: The inactivation effect of sterilization method using 400 nm of UVA-LED upon Pseudomonas aeruginosa and Staphylococcus aureus has been verified and it is considered as a useful method in inactivating the contact lenses.

Comparative Study on Some Factors Affecting the Oxyfluorfen Inactivation in Soil (Oxyfluorfen의 토양중(土壤中) 불활성화(不活性化)에 관여(關與)하는 수종요인(數種要因)의 비교연구(比較硏究))

  • Kim, D.K.;Lee, J.M.;Guh, J.O.;Lee, K.
    • Korean Journal of Weed Science
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    • v.6 no.2
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    • pp.146-153
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    • 1986
  • The study was conducted to estimate the oxyfluorfen inactivation proceeding which have or have not organism and soil microorganism in soil by biological testing method, under the different condition of soil class and temperature. The results obtained in this experiment are as follows: 1. Under the condition of high temperature, chemical`s inactivation resulted in early and the extent of it`s inactivation was increased. 2. The extent of inactivation was more rapid in the sandy soil than in the clay. 3. Among the soil class, the differentiation of the day of demand with inactivation at the 50% and 95% was significantly increased low temperature period. 4. In sandy soil, inactivation`s differentiation by adding organism was found, but by adding microorganism was not found. 5. In clay, chemical's inactivation was increased rather by adding microorganism than by adding organism. 6. Among under the condition of soil class, by adding organism and micrrorganism, chemical`s inactivation (Probit growing period at 50% and 95%) was shorted. And among the soil class, it was shortter sandy soil than clay or silty loam soil.

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