• Title/Summary/Keyword: Isoenzymes

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Effects of Pseudomonas aureofaciens 63-28 on Defense Responses in Soybean Plants Infected by Rhizoctonia solani

  • Jung, Woo-Jin;Park, Ro-Dong;Mabood, Fazli;Souleimanov, Alfred;Smith, Donald L.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.379-386
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    • 2011
  • The objective of this work was to investigate the ability of the plant growth-promoting rhizobacterium Pseudomonas aureofaciens 63-28 to induce plant defense systems, including defense-related enzyme levels and expression of defense-related isoenzymes, and isoflavone production, leading to improved resistance to the phytopathogen Rhizoctonia solani AG-4 in soybean seedlings. Seven-day-old soybean seedlings were inoculated with P. aureofaciens 63-28, R. solani AG-4, or P. aureofaciens 63-28 plus R. solani AG-4 (P+R), or not inoculated (control). After 7 days of incubation, roots treated with R. solani AG-4 had obvious damping-off symptoms, but P+R-treated soybean plants had less disease development, indicating suppression of R. solani AG-4 in soybean seedlings. Superoxide dismutase (SOD) and catalase (CAT) activities of R. solani AG-4-treated roots increased by 24.6% and 54.0%, respectively, compared with control roots. Ascorbate peroxidase (APX) and phenylalanine ammonia lyase (PAL) activities of R. solani AG-4-treated roots were increased by 75.1% and 23.6%, respectively. Polyphenol oxidase (PPO) activity in soybean roots challenged with P. aureofaciens 63-28 and P+R increased by 25.0% and 11.6%, respectively. Mn-SOD (S1 band on gel) and Fe-SOD (S2) were strongly induced in P+R-treated roots, whereas one CAT (C1) and one APX (A3) were strongly induced in R. solani AG-4- treated roots. The total isoflavone concentration in P+Rtreated shoots was 27.2% greater than the control treatment. The isoflavone yield of R. solani AG-4-treated shoots was 60.9% less than the control.

Purification and Some Properties of Polyphenol Oxidase from Arrowroot (칡 뿌리의 Polyphenol Oxidase의 정제 및 성질에 관한 연구)

  • Oh, Man-Jin;Lee, Won-Yong;Lee, Ka-Soon
    • Applied Biological Chemistry
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    • v.31 no.4
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    • pp.331-338
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    • 1988
  • Acetone powder was prepared from raw arrowroots and the polyphenol oxidases of crude enzyme prepared from acetone powder were identified 5 isoenzymes by staining with catechol containing 0.05% phenylene diamine. The crude enzyme was passed through the columns of ion exchangers and gel permeation to fractionate the polyphenol oxidases. The main fraction of polyphenol oxidase appeared to be purified by 94-fold, with the activity yield of 45.4%, and its molecular weight was determined as 38,500 by poly acrylamide gel electrophoresis. The optimal pH and temperature for the enzyme activity were pH 7.5 and $50^{\circ}C$, respectively. The purified enzyme showed a high affinity for catechol and pyrogallol. The Michaelis constant for catechol was calculated to be 16.67mM according to the Lineweaver-Burk method. The enzyme activity was strongly inhibited by L-ascorbic acid, sodium bisulfite, EDTA and KCN, and totally inhibited, by $Fe^{3+}$ at a concentration of 1mM. However the enzyme was activated by $Zn^{2+}$ approximately 1.7 times at the same concentration.

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Effects of 6-Shogaol, A Major Component of Zingiber officinale Roscoe, on Human Cytochrome P450 Enzymes in vitro (생강의 주성분인 6-Shogaol이 인체 약물대사효소인 Cytochrome P450에 미치는 영향)

  • Kim, Jin
    • Korean Journal of Medicinal Crop Science
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    • v.24 no.1
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    • pp.7-13
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    • 2016
  • Background : Ginger has been extensively used in foods and traditional medicines in Asian countries. Despite its frequent consumption in daily life, the mechanism of potential interactions between ginger components-drug has not been examined. To elucidate the mechanism of governing the effects of 6-shogaol, a primary constituent of dried ginger, on human cytochrome P450 (CYP) isoenzymes an incubation studies were carried out using pooled human liver microsome (HLM). Methods and Results : CYP isoenzyme specific substrate was incubated with multiple concentrations of inhibitor, HLM and cofactors. 6-shogaol showed a potent inhibitory effect on CYP2C9, CYP1A2 and CYP2C19 with half maximal inhibitory concentration ($IC_{50}$) values of 29.20, 20.68 and $18.78{\mu}M$ respectively. To estimate the value of the inhibition constant ($K_i$) and the mode of inhibition, an incubation study with varying concentrations of each CYP isoenzyme-specific probe was performed. 6-shogaol inhibited CYP2C9 and CYP2C19 noncompetitively ($K_i=29.02$ and $19.26{\mu}M$ respectively), in contrast, the inhibition of CYP1A2 was best explained by competitive inhibition ($K_i=6.33{\mu}M$). Conclusions : These findings suggest that 6-shogaol may possess inhibitory effects on metabolic activities mediated by CYP1A2, CYP2C9 and CYP2C19 in humans.

Chlorophyll Fluorescence and Antioxidative Enzyme Activity of Crinum Leaves Exposed to Natural Environmental Stress in Winter (겨울철 자연환경에 노출된 문주란 잎의 엽록소형광과 항산화효소 활성에 관한 연구)

  • 오순자;고석찬
    • Korean Journal of Environmental Biology
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    • v.22 no.1
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    • pp.233-241
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    • 2004
  • Chlorophyll fluorescence and antioxidative enzyme activity were investigated from leaves of Crinum asiaticum var. japonicum under the natural condition in winter, in order to monitor plant response and physiological states such as vitality, productivity and so on. In the O-J-I-P transients, the fluorescence intensity of J, I, P-step decreased remarkably depending on temperature drop in winter. The photochemical efficiencies of PSII, Fv/Fm, were significantly low in late winter with decrease of Fm. These results indicate that Crinum plants were affected by seasonal drop of temperature. The catalase activity significantly decreased depending on temperature drop in winter. However, the activity of superoxide dismutase ascorbate peroxidase and peroxidase slightly increased in winter while some isoenzymes appeared in winter. These results, with the remarkable decrease of Ev/Fm in winter, represent that Crinum plants were exposed to oxidative stress and subsequently damaged leading to cell death.

Correlation of hypoechogenic lesions with lactic dehydrogenase isoenzymes, culture and cytological findings of prostatic fluid and bilpsied tissue in dogs (개에서 전립선액과 생검조직에 대한 젖산탈수소 효소치, 배양 및 세포학적 검사와 초음파상의 저에코 영역과의 연관성)

  • Eom, Ki-dong;Yoon, Jung-hee;Sung, Jai-ki
    • Korean Journal of Veterinary Research
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    • v.38 no.3
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    • pp.664-673
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    • 1998
  • 초음파학적 검사에 따라 미만성 저에코 영역, 낭포, 다병소성 낭포 및 비후된 피막 등 비정상적 소견을 보이는 전립선에서 추출한 전립선액과 생검조직에서의 LDH 분획비(LDH I/V)의 평균치는 각각 $0.92{\pm}0.55$, $2.69{\pm}0.82$, 정상군(각각 $0.23{\pm}0.20$, $0.57{\pm}0.36$)에 비하여 유의성(p<0.01) 있게 높았다. 세포병리학적 검사에서 전립선 비대증 및 전립선염으로 나타난 비정상군($2.76{\pm}0.77$의 생검조직 LDH 평균분획비는 정상군($1.38{\pm}1.19$)에 비해 유의성 있게 높았으나 전립선액에서는 분획비의 유의차가 인정되지 않았다. 배양결과에 따른 전립선액 및 조직시료의 LDH 분획비는 정상군과 비정상군 사이에서의 유의성은 인정되지 않았다. 결론적으로 저에코 영역 및 낭포부위는 생검조직과 전립선액의 배양 및 세포병리학적 검사에 따른 비정상성 및 높은 LDH 분획비와 밀접한 연관성을 보였다. 특히 경직장 초음파상에 나타나는 미만성 저에코 영역은 전립선 비대증 및 만성 전립선염 등과 같은 병적 관련성을 내포하고 있는 것으로 사료된다.

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Establishment and Characterization of the Fibroblast Line from Silkie Bantam

  • Li, L.F.;Guan, W.J.;Li, H.;Bai, X.J.;Ma, Y.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.4
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    • pp.492-499
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    • 2009
  • A Silkie Bantam embryo fibroblast line (named SBF59 line) was successfully established by using direct explant culture and cryopreservation techniques. Cell morphology, viability, dynamic growth and contamination were tested and the karyotype and levels of isoenzymes of lactic dehydrogenase and malic dehydrogenase were analyzed. Four kinds of fluorescent protein extrogenes, including $pEGFP-N_3$, $pECFP-N_1$, $pEYFP-N_1$ and $pDsRed1-N_1$ were transfected into the cells. The results showed that the cells were healthy and possessed a fibrous structure without a change in morphology. The average viability of the cells was 96% before freezing and 90.5% after thawing. The growth curve appeared as typical "S" shape and the cell growth passed through a detention phase, a logarithmic phase and a platform phase; the estimated population doubling time (PDT) was 38.5 h; assays for the presence of bacteria, fungi, viruses and mycoplasmas were negative; the cell line showed no cross contamination when assessed by isoenzyme analysis; the chromosome number was 2n = 78 on more than 88% of occasions; four kinds of fluorescent protein extro-genes appeared to be expressed effectively with a high transfection efficiency between 18.3% and 42.3%. The cell line met the required quality control standard. It not only preserves the genetic resources of the important Silkie Bantam at the cellular level but also provides valuable materials for genomic, post-genomic, somatic cell cloning research and other applications.

Purification and Properties of Extracellular Inulinase of Pseudomouas sp. (Pseudomonas sp.가 생산하는 Inulinase에 관한 연구 -효소의 정제와 성질 -)

  • 이태경;최용진;양한철
    • Microbiology and Biotechnology Letters
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    • v.16 no.4
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    • pp.259-264
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    • 1988
  • Two forms of extracellular inulinase, designated as PI and PII were detected in the crude enzyme preparation from n species of Pseudomonas isolated from soil. PI and PII were purified to homogeneity by ammonium sulfate fractionation, DEAE Sephadex A-50 chromatography, Sephadex G-100 and Sephadex G-200 gel filteration. Both isoenzymes catalyzed specifically and endowise the cleavage of the $\beta$-2,1-fructofranoside linkage of inulin, and displayed no action upon sucrose, raffinose and levan. The optimal pH values for the PI and PII enzyme were pH 5.5 and 6.0, respectively and the highest activity of the two enzymes was observed at 55$^{\circ}C$. The Km values of PI and PII were calculated to be 2$\times$10$^{-3}$M and 5$\times$10$^{-3}$M, respectively.

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Drug Interaction in New Antipsychotics (새로운 항정신병약물의 약물상호작용)

  • Kim, Yong Sik;Kang, Ung Gu;Roh, Myoung Sun
    • Korean Journal of Biological Psychiatry
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    • v.7 no.1
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    • pp.14-20
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    • 2000
  • Recently atypical antipsychotics have been used as first line agent in the treatment of schizophrenia, and also played a significant role in the treatment of many kinds of psychiatric disorders. The pharmacokinetic and pharmacodynamic properties of these newer antipsychotics are well known through preclinical and early clinical trials. However, it is important to note the limitations of the results due to its relatively short experience. Clozapine is eliminated principally by the hepatic P450 1A2 and 3A4 cytochrome enzymes. 1A2 inducers such as carbamazepine and smoking can reduce its half-life, while 1A2 inhibitors such as SSRIs, especially fluvoxamine can increase its duration of action. Carbamazepine should be avoided in a patient on clozapine because of carbamazepine's potential effects on bone marrow. Benzodiazepines tend to increase the chances of sedation, delirium and respiratory depression. Risperidone is metabolized to 9-hydroxyriperidone by the hepatic P450 2D6 cytochrome enzymes. Fluoxetine and paroxetine, 2D6 inhibitors interfere with metabolism, but 9-hydroxyrisperidone has similar biological activity as parental drug, so it has little affect on the outcome. Olanzapine shows minimal capacity to inhibit cytochrome P450 isoenzymes and shows minimal chance of drug interaction. It is eliminated principally by the hepatic P450 1A2 and 2D6 cytochrome enzymes.

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Biochemical Changes Induced due to Staphylococcal Infection in Spongy Alphonso Mango(Mangifera indica L.) Fruits

  • Janave, Machhindra Tukaram
    • Journal of Crop Science and Biotechnology
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    • v.10 no.3
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    • pp.167-174
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    • 2007
  • Spongy Alphonso mangoes were found to be infected with Staphylococcus bacteria. A Gram positive Staphylococcus strain was isolated from spongy pulp and identified from CABI Bioscience, UK, by partial 16S rDNA sequence analysis and by morphological and biochemical characterization through IMTECH, Chandigarh, India. Although identification by both of these methods indicated the organism belonged to same genus, different species names were given. Changes in total phenolics, reducing, and non-reducing sugars, respiration rate, total carotenoids, peroxidase(POX), and catalase activities were monitored during ripening of these fruits. The climacteric rise in spongy fruits was marked by an increase in respiration rate and a decrease in sugar content. Total phenolics content increased in spongy fruits as compared to ripe non-spongy fruits. Development of corky white tissue in spongy fruits was associated with about a 2.5-fold reduction in total carotenoids and a concomitant increase in lipoxygenase-mediated, $\beta$-carotene co-oxidation. A marked decrease in soluble protein content and about a 1.5-fold increase in POX activity was observed. Maximum POX activity was confined to 50-70%$(NH_4)_2SO_4$ fraction. The intense dark bands visible after POX specific substrate staining of the Native gel indicated a high expression of isoenzymes of POX in spongy fruits. Similarly, changes in levels of catalase activity were also observed in spongy fruits. The results suggest that infection of Alphonso mangoes with Staphylococcus bacteria affects the normal ripening processes of the fruit interfering with the carbohydrate and carotenoid metabolism. Also, the studies indicate the expression of POX and catalase enzymes as a plant defense response to microbial invasion.

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Association of a Pyruvate Kinase M2 (PKM2) Polymorphism with Back Fat Thickness in Berkshire Pigs

  • Cho, Eun-Seok;Jeon, Hyeon-Jeong;Lee, Si-Woo;Park, Jong-Woon;Raveendar, Sebastian;Jang, Gul-Won;Kim, Tae-Hun;Lee, Kyung-Tai
    • Journal of Animal Science and Technology
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    • v.55 no.6
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    • pp.515-520
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    • 2013
  • Pyruvate kinase M2 (PKM2) is a key regulatory enzyme in the glycolytic pathway. It is one of four pyruvate kinase isoenzymes that widely differ in their occurrence according to tissue type. PKM2 is expressed in differentiated tissues, such as fat tissues, lung, as well as normal proliferating cells, embryonic cells, and tumor cells. The objective of this study was to investigate the association of single nucleotide polymorphisms (SNPs) in the PKM2 gene with meat quality traits in Berkshire pigs. We detected a SNP (g.34341 A>G) in the 3'UTR region of the PKM2 gene in 670 Berkshire pigs through DNA sequencing. Three genotypes, AA, AG, and GG, were found for this SNP, but based on an association analysis with meat quality traits, genotype AA was significantly associated with thicker back fat than genotype GG (p=0.027). Therefore, the g.34341 A>G polymorphism in the 3'UTR region of the porcine PKM2 gene could be applied in pig breeding programs to improve back fat thickness.