• Title/Summary/Keyword: primary metabolism

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Characterization of Laccase Purified from Korean Pycnoporus cinnabarinus SCH-3 (한국산 주걱송편버섯(Pycnoporus cinnabarinus) SCH-3로부터 정제 된 Laccase의 특성)

  • Park, Eun-Hye;Yoon, Kyung-Ha
    • The Korean Journal of Mycology
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    • v.31 no.2
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    • pp.59-66
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    • 2003
  • Laccase produced by Pycnoporus cinnabarinus SCH-3 isolated from Korea was partially purified using ultrafiltration, anion exchange chromatography and affinity chromatography, The laccase was produced as the predominant extracellular phenoloxidase during primary metabolism. Neither lignin peroxidase nor manganese-dependent peroxidase were detected in the culture fluid. In order to examine the effect of inducers in laccase production, 2,5-xylidine was added in the culture of Pycnoporus cinnabarinus SCH-3. Addition of 2,5-xylidine enhanced 25-fold laccase production. Purified laccase was a single polypeptide having a molecular mass of approximately 66 kDa, as determined by SDS-polyacrylamide gel electrophoresis, and carbohydrate content of 9%. $K_{m}\;and\;V_{max}$ values for laccase with ABTS [2,2-azinobis (3-ethylbenzthiazoline 6-sulfonic acid)] as a substrate (Lineweaver-Burk plot) was determined to be $44.4{\mu}M\;and\;56.0{\mu}mole$, respectively. The optimal pH for laccase activity was found to be 3.0. The enzyme was very stable for 1 hour at $60{\circ}C$. Half-life ($t_{1/2}$) of the enzyme was about 10 min at $80{\circ}C$. Spectroscopic analysis of purified enzyme indicated that the enzyme was typical of copper-containing protein. Substrate specificity and inhibitor studies for laccase also indicated to be a typical fungal laccase. The N-terminal amino acid sequence of the P. cinnabarinus SCH-3 laccase showed 94% of homology to the N-terminal sequences of laccases from P. cinnabarinus PB and P. coccineus.

Genes of Wild Rice (Oryza grandiglumis) Induced by Wounding and Yeast Extract (상처와 효모추출물 처리조건에서 유발되는 야생벼 유전자 스크린)

  • Shin, Sang-Hyun;Im, Hyun-Hee;Lee, Jai-Heon;Kim, Doh-Hoon;Chung, Won-Bok;Kang, Kyung-Ho;Cho, Sung-Ki;Shin, Jeong-Sheop;Chung, Young-Soo
    • Journal of Life Science
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    • v.14 no.4
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    • pp.650-656
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    • 2004
  • Oryza grandiglumis (CCDD, 2n=48), one of the wild rice species, has been known to possess fungal-,bacterial-, and insect-resistance against sheath blight, rice blast, bacterial leaf blight and brown plant hopper (Nilaparvata lugens). To rapidly isolate differentially expressed genes responding to fungal and wounding stress, wounding and yeast extract were treated to O. grandiglumis for 24 hrs. Suppression subtractive hybridization (SSH) method was used to obtain differentially expressed genes from yeast extract and wounding treated plants. Seven hundreds and seventy six clones were obtained by subcloning PCR product, and colony array and screening were carried out using radio-isotope labeled cDNA probes prepared from the wounding and yeast extract treated plants. One hundred and fifteen colonies were confirmed as true positive ones. Average insert size of the clones were ranged from 400 bp to 700 bp and all the inserts were sequenced. To decide the identity of those clones, sequences were analyzed by sequence homology via GenBank database. The homology search result showed that 68 clones were matched to the genes with known function; 16 were related to primary metabolism, 5 to plant retrotransposons, 5 to defense related metallothionein-like genes. In addition to that, others were matched to various genes with known function in amino acid synthesis and processing, membrane transport, and signal transduction, so on. In northern blot analysis, induced expressions of ogwfi-161, ogwfi-646, ogwfi-663, and ogwfi-695 by wounding and yeast extract treatments were confirmed. The result indicates that SSH method is very efficient for rapid screening of differentially expressed genes.

The Effects of Cooperative Learning to Study the Unit 'Metabolism' in High School: Application of STAD Model (고등학교 생물 '물질대사' 단원에서 협동학습의 효과: STAD 모형의 적용)

  • Chung, Young-Lan;Lee, Hye-Won
    • Journal of The Korean Association For Science Education
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    • v.23 no.1
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    • pp.35-46
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    • 2003
  • Problem solving ability, having been thought as one of the most important goals of science education is also a primary task for the current education. Indeed, the students' problem solving ability has shown almost no actual progress, despite our long accumulated science education. Under this circumstances, cooperative learning, a way to grow students' positive inter-dependence and problem solving ability in the basis of their active participation and discussion, was proposed as an effective teaching method. But, results have not consistently shown the advantage of cooperative learning over traditional learning for promoting academic achievement in science. Studies have consistently shown greater effectiveness on affective aspects. But, relatively few have focused on biology in Korea. The purpose of this study was to examine the effects of cooperative learning on the achievement and attitude of high school biology students. The pretest-posttest control group design was applied. The sample consisted of 50 11th-grade female students in experimental group(cooperative learning Student Team Achievement Division model) and 50 students in control group(traditional learning). Students in both groups recieved identical content instruction on the unit 'II. Methabolism'. These groups were treated for 13 hours during 5 weeks. Achievement data were collected using a 24-item multiple-choice test(content validity= .85). Science attitude was measured by an instrument which adapted by Kim In Hee(1994). The instrument(Cronbach $\alpha$=.89) included 40 items in four subscales: attitude toward science, social meaning of science, attitude toward science class, and scientific attitude. An analysis of covariance (ANCOVA) was used as the data analysis procedure. For the achievement data, no significant difference exists between the cooperative and traditional groups (p> .05). But, cooperative learning was effective in low-ability students(p < .05). For the science learning attitude data, cooperative learning was more effective than the traditional one(p< .05). Students in the cooperative group acheived better than those in traditional one especially in the subscale of attitude toward science class. There was no meaningful difference of the two methods in both high and average ability students, while cooperative learning was more effective than the traditional one in low ability students(p<.05).

Polymorphisms of GSTM1 and CYP1A1, and Susceptibility to Primary Lung Cancer in Korean Males (한국인 남성에서 GSTM1과 CYP1A1 유전자 다형성과 원발성폐암의 유전적 감수성)

  • Bae, Nack-Cheon;Lee, Su-Yeon;Chae, Po-Hee;Kang, Kyung-Hee;Kim, Kyung-Rock;Cha, Seung-Ick;Chae, Sang-Chul;Kim, Chang-Ho;Jung, Tae-Hoon;Park, Jae-Yong
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.5
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    • pp.568-578
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    • 2001
  • Background : Lung cancer is frequently cited as an example of a disease caused solely by exposure to environmental carcinogens. However, there is a growing realization that the genetic constitution is also important in determining individual's susceptibility to lung cancer. This genetic susceptibility may result from functional polymorphims of the genes involved in carcinogen metabolism. In this study, the association between GSTM1 and CYP1A1 polymorphisms and the lung cancer risk in Korean males was investigated. Materials and Method : The study population consisted of 153 male lung cancer patients and 143 healthy male controls. The GSTM1 and CYP1A1 genotypes were determined by multiplex PCR and PCR-RFLP analysis. Result : There were no significant differences in the frequency of the GSTM1 null genotype between the cases and the controls. When the cases were categorized by their histologic type, the frequency of the GSTM1 null genotype in the small cell carcinoma group was higher than those of the controls(67.2% vs 55.9%), but the difference was not statistically significant(OR=1.772 ; 95% CI=0.723-4.340). The distribution of the CYP1A1 MspI genotypes among the cases were similar to those among the controls. When the cases were grouped by their histologic type, the ml/m1, ml/m2, m2/m2 genotypes frequencies among the small cell carcinomas(23.0%, 38.5%, and 38.5%, respectively) were significantly different from those of the controls(36.4%, 46.2%, and 7.4%, respectively, p<0.05). When the m1/m1 genotype was used as a reference, the ml/m2 and m2/m2 genotypes were associated with an increased risk for small cell lung cancer(ml/m2 genotype : OR=1.337, 95% CI=0.453-3.947 ; m2/m2 genotype : OR=3.374, 95% CI=1.092-10.421). Conclusion : These results suggest that the GSTM1 and CYP1A1 genotypes may be a genetic determinant of the risk for lung cancer, particlulary small cell carcinoma. Further investigation is needed to confirm these results.

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