• Title/Summary/Keyword: L-malate

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Substantial Protective Immunity Conferred by a Combination of Brucella abortus Recombinant Proteins against Brucella abortus 544 Infection in BALB/c Mice

  • Arayan, Lauren Togonon;Huy, Tran Xuan Ngoc;Reyes, Alisha Wehdnesday Bernardo;Hop, Huynh Tan;Son, Vu Hai;Min, WonGi;Lee, Hu Jang;Kim, Suk
    • Journal of Microbiology and Biotechnology
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    • v.29 no.2
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    • pp.330-338
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    • 2019
  • Chronic infection with intracellular Brucella abortus (B. abortus) in livestock remains as a major problem worldwide. Thus, the search for an ideal vaccine is still ongoing. In this study, we evaluated the protective efficacy of a combination of B. abortus recombinant proteins; superoxide dismutase (rSodC), riboflavin synthase subunit beta (rRibH), nucleoside diphosphate kinase (rNdk), 50S ribosomal protein (rL7/L12) and malate dehydrogenase (rMDH), cloned and expressed into a pMal vector system and $DH5{\alpha}$, respectively, and further purified and applied intraperitoneally into BALB/c mice. After first immunization and two boosters, mice were infected intraperitoneally (IP) with $5{\times}10^4CFU$ of virulent B. abortus 544. Spleens were harvested and bacterial loads were evaluated at two weeks post-infection. Results revealed that this combination showed significant reduction in bacterial colonization in the spleen with a log protection unit of 1.31, which is comparable to the average protection conferred by the widely used live attenuated vaccine RB51. Cytokine analysis exhibited enhancement of cell-mediated immune response as IFN-${\gamma}$ is significantly elevated while IL-10, which is considered beneficial to the pathogen's survival, was reduced compared to control group. Furthermore, both titers of IgG1 and IgG2a were significantly elevated at three and four-week time points from first immunization. In summary, our in vivo data revealed that vaccination with a combination of five different proteins conferred a heightened host response to Brucella infection through cell-mediated immunity which is desirable in the control of intracellular pathogens. Thus, this combination might be considered for further improvement as a potential candidate vaccine against Brucella infection.

Proteome Characterization of Sorghum (Sorghum bicolor L.) at Vegetative Stage under Waterlogging Stress (토양 과습 조건하에서 수수 잎의 단백질 양상)

  • Yun, Min Heon;Jeong, Hae-Ryong;Yoo, Jang-Hwan;Roy, Swapan Kumar;Kwon, Soo-Jeong;Kim, Joo-Ho;Chun, Hyen Chung;Jung, Ki Yuol;Cho, Seong-Woo;Woo, Sun-Hee
    • Korean Journal of Plant Resources
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    • v.31 no.2
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    • pp.124-135
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    • 2018
  • The study was performed to explore the molecular changes in the vegetative stage (3-and 5-leaf) of sorghum under waterlogging stress. A total of 74 differentially expressed protein spots were analyzed using LTQ-FT-ICR MS. Among them, 12 proteins were up-regulated and 3 proteins were down-regulated. Mass spectrometry (MS) results showed that about 50% of the proteins involved in various metabolic processes. The level of protein expression of malate dehydrogenase and glyceraldehyde-3-phosphate dehydrogenase related to carbohydrate metabolic process increased in both 3 and 5-leaf stage under waterlogging stress. These proteins are known to function as antistress agents against waterlogging stress. The expression of oxygen-evolving enhancer protein 1 protein related to photosynthesis was slightly increased in the treated group than in the control group, however the expression level was increased in the 5-leaf stage compared to the 3-leaf stage. Probable phospholipid hydroperoxide glutathione peroxidase protein and superoxide dismutase protein related to response to oxidative stress showed the highest expression level in 5-leaf stage treatment. This suggests that the production of reactive oxygen species by the waterlogging stress was the most abundant in the 5-leaf treatment group, and the expression of the antioxidant defense protein was increased.

Metabolic Gene Expression in Lipid Metabolism during Cotyledon Development in Cucumbers and the Possibility of a Secondary Transport Route of Acetyl Units (오이 떡잎의 발달에서 지방 대사관련 유전자의 발현과 아세틸 단위체의 2차 경로 가능성)

  • Cha, Hyeon Jeong;Kim, Dae-Jae
    • Journal of Life Science
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    • v.24 no.10
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    • pp.1055-1062
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    • 2014
  • We investigated the expression of cucumber genes involved in lipid mobilization and metabolism during cotyledon development to compare gene activity and to study the direction of carbon (acetyl unit) transport between glyoxysomes and mitochondria. The core metabolic pathway involving 10 genes was examined in four intracellular compartments: glyoxysomes (peroxisomes), mitochondria, chloroplasts, and cytosol. Additionally, we tested the early germination response of dark-grown seedlings and the immediate light response for a further 3 days. According to the reverse transcription polymerase chain reaction (RT-PCR), 3-L-ketoacyl-CoA thiolase 2 (Thio2), isocitrate lyase (ICL), and malate synthase (MS), the genes involved in storage lipid mobilization showed a similar and consistent pattern of gene expression in seedling development. Furthermore, coordinate expression of the A BOUT DE SOUFFLE (BOU) gene with ICL and MS during seedling emergence pointed to a possible secondary route of acetyl unit (acetyl-CoA) transport between peroxisomes and mitochondria in cucumber. The expression of the BOU gene was light dependent, as shown by BOU activity in Arabidopsis, suggesting that the dark condition also results in weak membrane biogenesis. In addition, several genes were active throughout the development of the green cotyledon, even during senescence. In conclusion, this study summarizes oil-seed germination and gene expression during cucumber cotyledon development and proposes an additional route for acetyl unit transport.

Radio-sensitivity of Dark-striped Field Mice, Apodemus agrarius, as a Biological Dosimeter in Radio-ecological Monitoring System (환경 방사선 생물학적 감시 지표로서 야생 등줄쥐의 방사선 감수성)

  • Kim, Hee-Sun;Nishimura, Y.;Kim, Chong-Soon
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.4 no.1
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    • pp.25-32
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    • 2006
  • This study examined the possibility of using dark-striped field mice as a biological indicator for the environmental radio-surveillance. For this study, dark-striped field mice were caught from five areas of Kyonggi, Kyongsang, Chungchong and Cholla provinces. The external morphological characteristics and isoenzymic types of dark-striped field mice were studied after they were captured. Among the external morphological characteristics, the dark-brown coat, dark back stripe, head-to-tail length, tail length, and ear length matched the taxonomical characteristics of dark-striped field mice. The analyses on L-lactate dehydrogenase, aspartate aminotransferase, and malate dehydrogenese revealed that one species of dark-striped field mice, called Apodemus agrarius, was inhabitated throughout a wide range of Korea. On the other hand, the frequency of micronuclei in peripheral polychromatic erythrocytes to survived mice after irradiation also analyzed. The LD50/30 of A. cgrarius and ICR mice were approximately 5 Gy and 7.9Gy, respectively. The results of the study reveal that wild A. asrarius have a high potential as a biological monitoring system to determine the impact of radiation in areas such as those within the vicinity of nuclear power plants.

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Deacidification Effect of Campbell Early Must via Carbonic Maceration : Effect of Enzyme Activity Associated with Malic-Acid Metabolism (Carbonic Maceration처리에 의한 Campbell Early 발효액의 감산 효과: 사과산 대사 관련 효소활성의 영향)

  • Chang, Eun-Ha;Jeong, Seok-Tae;Jeong, Sung-Min;Roh, Jeong-Ho;Park, Kyo-Sun;Park, Seo-Jun;Choi, Jong-Uck
    • Food Science and Preservation
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    • v.18 no.5
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    • pp.795-802
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    • 2011
  • To determine the deacidification factor during carbonic maceration (CM), different temperature conditions were studied. The pH was higher in CM-$35^{\circ}C$ and CM-$25^{\circ}C$ and was lower in CM-$45^{\circ}C$. The total acid was inversely related to the pH. The malic-acid level decreased much more in CM-$35^{\circ}C$ than in CM-$45^{\circ}C$ while the lactic-acid level increased much more in CM-$35^{\circ}C$. The activity of the NADP-malic enzyme, which catalyzes the oxidative decarboxylation of L-malate into pyruvate, $CO_2$, and NADPH, was higher in CM-$25^{\circ}C$ and CM-$35^{\circ}C$ while CM-$45^{\circ}C$ showed no NADP-malic enzyme activity. The malic-dehydrogenase (MDH) activity was higher in CM-$25^{\circ}C$ and CM-$35^{\circ}C$ while CM-$45^{\circ}C$ showed no MDH activity. The oxalacetate decarboxylase activity was similar to the NADP-malic-enzyme and MDH activities. Pyruvate decarboxylase activity was shown in all the CM treatments. The L-lactic dehydrogenase (LDH) activity was not explored in the fermentation of pyruvate to lactate via LDH in the grapes during CM. In this study, it was confirmed that carbonic maceration reduced the malic acid during fermentation and was affected by the temperature. Moreover, it was assumed that the deacidification during the carbonic maceration of the grapes was probably correlated with the degradation enzyme activity of malic acid.