• Title/Summary/Keyword: mixed enzyme

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Effects of Enzyme Inducers and Glutathione on the Embryotoxicity of Cyclophosphamide in Cultured Rat Embryos (효소유도제 및 glutathione이 전배자배양된 랫드태자에서 cyclophosphamide의 독성에 미치는 영향)

  • 한순영;신재호;권석철;강명옥;이유미;김판기;양미라;박귀례;장성재
    • Toxicological Research
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    • v.11 no.1
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    • pp.31-36
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    • 1995
  • Cyclophosphamide (CP) must be enzymatically activated by cytochrome P450(CYP)-linked mixed-function oxidation pathway to be either mutagenic or teratogenic. Influences of alterations in hepatic mixed-function oxidase acitivity and glutathione (GSH) content on the embryotoxicity of CP were studied in rat whole embryo culture system. The embryotoxicity of CP was compared using rat S-9 fraction (S-9) pretreated with chemicals inducing different CYP isozymes, acetone (ACE), Aroclor 1254 (ARO), $\beta$-naphthoflavone (NAF) and phenobarbital (PHE). When 10.5 day embryos were cultured in the immediately centrifuged rat serum for 48 hrs using general gas char{ging schedule, CP$(40{\mu}g/ml)$ with S-9 induced by either NAF or PHE increased the incidence of realformations and significantly decreased embryonic growth compared with the non-induced S-9 group. ACE or ARO induced S-9 group showed no significant difference in embryonic growth. These data suggest that PB and/or NAF inducible CYP isoenzymes are mainly involved in the activation of CP. To examine the effect of GSH on the embryotoxicity of CP, 10.5 day embryos were exposed to CP and S-9 after preincubation with 10 mM of GSH for 3 hrs. In the GSH pretreated group the growth of embryos increased significantly compared with that of the untreated group, suggesting that GSH may protect embryos in culture from some toxic effects of CP.

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Trypsin Inhibitor from Streptomyces sp. (Part 2) Biological Activities or the Inhibitor (Streptomyces 속 균주가 생성하는 Trypsin Inhibitor (제2보) 저해물질의 생물학적 작용상)

  • Yi, Dong-Heui;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.10 no.4
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    • pp.283-288
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    • 1982
  • Trypsin inhibitor produced by Streptomyces sp. was investigated its reactive characteristics against trypsin. The mode of inhibition against trypsin was mixed type of non-competitive and competitive with casein, and enzyme-inhibitor complex was formed rapidly. The inhibitory activity was increased by the addition of isoleucine and depressed by silver, mercuric or cupric ion. And when egg albumin or hemoglobin was used as substrate for trypsin, the inhibition ratio was changed. The inhibitor inhibited coagulation of blood of bovine.

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Purification and enzymatic characteristics of myrosinase from radish (무에서 추출한 myrosinase의 정제 및 효소학적 특성)

  • Shim, Ki-Hwan;Kang, Kap-Suk;Seo, Kwon-Il
    • Applied Biological Chemistry
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    • v.36 no.2
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    • pp.86-92
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    • 1993
  • Myrosinase from radish was purified by DEAE Bio-Gel, Con-A, and Superose-6 column. The purified myrosinase(II) possessed 2 subunits, and their molecular as determined by SDS-polyacrylamide gel electrophoresis were 53 and 39 KD, respectively. The specific activity of purified enzyme was 37,500 units/mg. The enzyme was purified approximately 44-fold compared to the crude enzyme. Optimum pH of the myrosinase was $6.5{\sim}7.0$ in phosphate and Tris-HCl buffer solutions. Optimum temperature of the enzyme was $37{\sim}38^{\circ}C$. The enzyme was stable at pH 7.0, and less than $30^{\circ}C$. Cu or Hg ion significantly inhibited the enzyme activity, but ascorbic acid enhanced, resulting in a maximum activity by 1 mM ascorbic acid. Among the ascorbic acid analogues, dehydroascorbic acid did not affect, whereas others showed a little effect, but less than ascorbic acid itself. Individual 2-mercaptoethanol and dithiothreitol (reducing agents) did not enhance the enzyme activity. but 2-mercaptoethanol effect was enhanced when mixed with ascorbic acid.

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Current Characteristics of a Flow Injection Type Enzyme-Sensor as the Variables of a Buffer Velocity, an Enzyme-Substrate Reaction and an Electrode for the Control of a Fermentation Process (완충용액유속, 효소.기질 반응 및 전극봉 요인에 따른 발효공정 제어용 흐름주입식 효소센서의 전류값 특성)

  • Song, Dae-Bin;Jung, Hyo-Seok;Kim, Sung-Tae
    • Journal of Biosystems Engineering
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    • v.32 no.6
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    • pp.455-461
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    • 2007
  • The electric current of a flow injection type enzyme-sensor was measured to confirm the stable operating conditions of the sensor. The current of the sensor was decreased as the buffer solution velocity increased. Under the limitation of the cycle time to be below 10 minutes, the effective ranges of the buffer solution velocity were suggested $0.10{\sim}0.26$, $0.12{\sim}0.24$, $0.1{\sim}0.25$ and $0.05{\sim}0.10\;cm/s$ of 1.0, 1.4, 2.4 and 3.4 mm of the electrode diameters, respectively. As the reaction time of the enzyme and the substrate was increased, the current was decreased because of the dilution between the sample and buffer solution. Therefore, it could be recommended that the reaction time was able to be selected as shortly as possible in consideration of the total cycle time. As the result of the experiments using a different volume ratio of the enzyme to substrate, it was concluded that the substrate had to be mixed with the same amount of the enzyme. The current have increased remarkably in proportion to the electrode diameter under 0.1 cm/s of the buffer solution velocity but there was no difference over 0.1 cm/s of the buffer solution velocity. The cross type arrangement of the electrode was highly suggested for application and machining of the sensor.

An Improved Method for Detection of Salmonella Typhi O Antigen with Staphylococcal Protein A Using Enzyme Immunoassay (포도구균의 A단백질을 이용한 효소면역법으로 살모넬라 O항원 검출)

  • Rhyu, Mun-Gan;Kim, Gum-Ryong;Lee, Choong-Ki
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.4
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    • pp.445-451
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    • 1987
  • Coagglutination method is widely used for the diagnosis of Salmonella infection. This test, however, has a disadvantage of false positive reaction due to the coagglutination of staphylococci with non-specific immune complexes or anti-staphylococci antibody in serum. Salmonell O antigen was detected by enzyme immunoassay with protein A-bearing Staphylococcus aureus as in the solid phase. Horse radish peroxidase was labeled to IgG specific against Salmonella O antigen. This enzyme immunoassay was much more sensitive than conventional coagglutination method without false poitive agglutination. To improve the sensitivity for detection of Salmonella O antigen in samples, we tried to determine the optimal concentration of normal IgG that inhibits non-specific binding of horse radish peroxidase labeled IgG to staphylococci, and to establish the optimal condition of reaction between antigen-antibody complex and staphylococci. Non-specific binding of horse radish peroxidase labeled specific IgG to staphylococci was almost blocked when the enzyme labeled IgG was 500-fold diluted with phosphate buffered saline containing 2mg/ml of normal IgG. When staphylococci coated with antibody to Salmonella O antigen were mixed with antigen-antibody complex and then incubated for 1 hour at room temperature, the minimal detectable concentration of Salmonella O antigen was 1ng/ml. The sensitivity of enzyme immunoassay was 100-fold greater than a conventional coagglutination method. This enzyme immunoassay could be expected as an improved method for detection of other infectious agents.

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Optimization of Maca (Lepidium meyenii) Extraction for Natural Beverage Development using Enzyme Treatment (효소처리에 의한 천연 마카음료 개발을 위한 최적 추출 조건)

  • Kim, Jeong-Ah;Im, Moo-Hyeog
    • Journal of the Korean Society of Food Culture
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    • v.34 no.3
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    • pp.361-368
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    • 2019
  • The purpose of this study was to establish the best optimized extraction condition for the optimal development of fresh maca beverage using low temperature extraction and enzyme treatment. Low temperatures were applied to prevent heat-related nutritional loss during the extraction process. Best extraction conditions were investigated based on the ratio of maca to water, the ratio of enzymes, extraction temperature and time, and agitation. The optimal enzyme conditions were also examined after the treatment of cellulase:pectinase mixture to maintain the original color and flavor, as well as to increase the extraction yield. When cellulase:pectinase was 1:1, the extraction rate ranged from 77.84 to 79.29%. In addition, the best extraction rate was found when maca was mixed with twice volume of water and incubated at $45^{\circ}C$ ($84.05{\pm}0.32%$) with 90 rpm ($87.13{\pm}0.46%$) agitation for 3 hours ($84.73{\pm}0.29%$). Furthermore, sensory evaluation showed a high score in flavor, sweetness, and overall acceptability after adding 3% jujube concentrate into a fresh maca beverage.

Purple Rice Extract Supplemented Diet Reduces DMH-Induced Aberrant Crypt Foci in the Rat Colon by Inhibition of Bacterial β-Glucuronidase

  • Summart, Ratasark;Chewonarin, Teera
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.749-755
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    • 2014
  • Background: Purple rice has become a natural product of interest which is widely used for health promotion. This study investigated the preventive effect of purple rice extract (PRE) mixed diet on DMH initiation of colon carcinogenesis. Materials and Methods: Rats were fed with PRE mixed diet one week before injection of DMH (40 mg/kg of body weight once a week for 2 weeks). They were killed 12 hrs after a second DMH injection to measure the level of $O^6$-methylguanine and xenobiotic metabolizing enzyme activities. Results: In rats that received PRE, guanine methylation was reduced in the colonic mucosa, but not in the liver, whereas PRE did not affect xenobiotic conjugation, with reference to glutathione-S-transferase or UDP-glucuronyl transferase. After 5 weeks, rats that received PRE with DMH injection had fewer ACF in the colon than those treated with DMH alone. Interestingly, a PRE mixed diet inhibited the activity of bacterial ${\beta}$-glucuronidase in rat feces, a critical enzyme for free methylazoxymethanol (MAM) release in the rat colon. These results indicated that purple rice extract inhibited ${\beta}$-glucuronidase activity in the colonic lumen, causing a reduction of MAM-induced colonic mucosa DNA methylation, leaded to decelerated formation of aberrant crypt foci in the rat colon. Conclusions: The supplemented purple rice extract might thus prevent colon carcinogenesis by the alteration of the colonic environment, and thus could be further developed for neutraceutical products for colon cancer prevention.

Effect of Enzyme Stabilization on Hardening of Clay-rock Brick (점토-골재 벽돌 경화에 있어 효소 사용의 효과)

  • Mitikie, Bahiru Bewket;Lee, Tai-Sik
    • Journal of the Korean Recycled Construction Resources Institute
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    • v.5 no.4
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    • pp.366-374
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    • 2017
  • This study investigates enzyme stabilization in clay-rock bricks through mechanical tests and image processing. Appropriate soil mixtures were designed using clay/crushed rock with ratios of 70/30, 60/40, 50/50, 40/60, and 30/70 by weight to verify the strength of the enzyme brick and soil compaction. The maximum compressive and flexural strengths in the 60/40 ratio mixture were found to be 5MPa and 1.25MPa, respectively; however, the maximum dry unit weight of $2.073g/cm^3$ was found in the 50/50 clay/gravel ratio mixture. Generally, the strength of the enzyme brick was improved by 27%. The paper concludes that in order to achieve optimal strength, soils should be mixed with the 60/40 clay/gravel ratio, which provides an adequate strength, while 50/50 ratio should be used for achieving more compaction. The SEM-EDX observation and Matlab image processing verified how the bond structure appeared after enzyme stabilization. It was found that enzymes created bond with the clay soil and the crushed rock for rendering strength and stability.

Activities of non-specific defense factors in cultured oblong rockfish(Sebastes oblongus) and rockfish(S. schlegeli) (양식 볼락류에서 비특이적 방어인자의 활성)

  • Kim, Jin-Do;Byun, Soon-Gyu;Park, Sung-Woo;Kim, Eun-Heui
    • Journal of fish pathology
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    • v.21 no.3
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    • pp.247-257
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    • 2008
  • To understand the activity of non-specific defence factors in cultured Sebastes, the antibacterial effect of the serum, skin mucus and homogenate of various organs from cultured oblong rockfish (Sebastes oblongus) and rockfish(Sebastes schlegeli) against pathogenic bacteria, Aeromonas hydrophila, Edwardsiella tarda, Vibrio anguillarum, and Streptococcus sp. was compared with that of flounder(Paralichthys olivaceus) and seabass(Leteolabrax japonicus). And the activities of proteolytic enzyme, chitinolytic enzyme and haemolycin as non-specific defence factor were investigated on the oblong rockfish and rockfish. Samples from oblong rockfish showed the highest antibacterial activity by lysoplate assay on agar plate mixed with pathogens, followed in descending order by rockfish, seabass, and flounder. Turbidimetric assay was carried to evaluate the lysozyme activity of fish samples against lyophilized cells of Micrococcus lysodeiktikus. The serum, kidney, liver, stomach, intestine and eyeball of oblong rockfish and the mucus and gill of rockfish appeared to have the highest lysozyme activity among the fish strains investigated. All samples except skin mucus, liver, and eyeball of oblong rockfish and rockfish showed proteolytic enzyme activity. Chitinolytic enzyme activity was showed in random sampling and haemolytic activity was remarkable in oblong rockfish. Therefore, Sebastes strain was proved to have effective defense mechanisms based on the antibacterial activities, and lysozyme, proteolytic enzyme, chitinolytic enzyme, and haemolycin were considered to act as the non-specific defence factor of Sebastes.

유사 생합성 경로를 가진 Streptomyces sp.의 혼합배양을 이용한 Doxorubicin 생합성

  • Choi, Yun-Hwa;Hong, Young-Soo;Lim, Jai-Yun;Lee, Jung-Joon
    • Microbiology and Biotechnology Letters
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    • v.25 no.6
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    • pp.580-585
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    • 1997
  • We selected two mutants namely strain D5 and Nu23 by mutagenesis of anthracycline producing Streptomyces: the former is an $\varepsilon$-rhodomycinone overproducing mutant selected from Streptomyces sp. C5, a baumycin producer and the latter, a blocked mutant of early pathway for doxorubicin biosynthesis obtained from Streptomyces peucetius ATCC 27952, a doxorubicin producer. The mutant strain Nu23 does not produce anthracycline metabolites but retains the most of enzyme activities converting aklavinone to doxorubicin and the mutant strain D5 produced $\varepsilon$-rhodomycinone at a level of 150 $\mu$g/ml. These strains were grown separately in NDYE medium and each was mixed at day 3 by equal volume of culture broth but the quantity of doxorubicin produced was far below an estimation based on the level of $\varepsilon$-rhodomycinone normaly produced by the strain D5. On the other hand doxorubicin was reached at maximum level after 4 days in the mixed culture condition which was composed of culture broth of strain D5 grown for 6 day and that of strain Nu23 grown for 3 day. It was turned out that the growth of mutant strain D5 was inhibited by the accumulation of daunorubicin and doxorubicin in mixed culture broth, which cause the limitation of $\varepsilon$-rhodomycinone.

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