• Title/Summary/Keyword: Metabolic enzymes

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Structural and Kinetic Characteristics of 1,4-Dioxane-Degrading Bacterial Consortia Containing the Phylum TM7

  • Nam, Ji-Hyun;Ventura, Jey-R S.;Yeom, Ick Tae;Lee, Yongwoo;Jahng, Deokjin
    • Journal of Microbiology and Biotechnology
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    • v.26 no.11
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    • pp.1951-1964
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    • 2016
  • 1,4-Dioxane-degrading bacterial consortia were enriched from forest soil (FS) and activated sludge (AS) using a defined medium containing 1,4-dioxane as the sole carbon source. These two enrichments cultures appeared to have inducible tetrahydrofuran/dioxane and propane degradation enzymes. According to qPCR results on the 16S rRNA and soluble di-iron monooxygenase genes, the relative abundances of 1,4-dioxane-degrading bacteria to total bacteria in FS and AS were 29.4% and 57.8%, respectively. For FS, the cell growth yields (Y), maximum specific degradation rate ($V_{max}$), and half-saturation concentration ($K_m$) were 0.58 mg-protein/mg-dioxane, $0.037mg-dioxane/mg-protein{\cdot}h$, and 93.9 mg/l, respectively. For AS, Y, $V_{max}$, and $K_m$ were 0.34 mg-protein/mg-dioxane, $0.078mg-dioxane/mg-protein{\cdot}h$, and 181.3 mg/l, respectively. These kinetics data of FS and AS were similar to previously reported values. Based on bacterial community analysis on 16S rRNA gene sequences of the two enrichment cultures, the FS consortium was identified to contain 38.3% of Mycobacterium and 10.6% of Afipia, similar to previously reported literature. Meanwhile, 49.5% of the AS consortium belonged to the candidate division TM7, which has never been reported to be involved in 1,4-dioxane biodegradation. However, recent studies suggested that TM7 bacteria were associated with degradation of non-biodegradable and hazardous materials. Therefore, our results showed that previously unknown 1,4-dioxane-degrading bacteria might play an important role in enriched AS. Although the metabolic capability and ecophysiological significance of the predominant TM7 bacteria in AS enrichment culture remain unclear, our data reveal hidden characteristics of the TM7 phylum and provide a perspective for studying this previously uncultured phylotype.

Protective Effect of Dietary Buchu (Allium tuberosum Rottler) on Oxidative Stress and Lipofuscin Formation in Streptozotocin-Induced Diabetic Rats (Streptozotocin-유발 당뇨쥐에서 부추식이의 산화적 스트레스 및 Lipofuscin 생성 억제 효과)

  • 이점옥;류승희;이유순;김정인;문갑순
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.8
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    • pp.1337-1343
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    • 2003
  • Diabetes mellitus has been known to be a state of increased oxidative stress. Free radical formation and lipid peroxidation are accelerated in this metabolic disorder. Buchu (Allium tuberosum Rottler) contains lots of antioxidative nutrients such as chlorophyll, vitamin C, $\beta$-carotene, phenolic compounds and sulfur compounds. To investigate the protective effects of buchu, 10% lyophilized buchu diet was fed to streptozotocin (STZ)-induced diabetic rats for 14 weeks and lipid peroxidation, protein oxidation, contents of reactive oxygen species, activities of antioxidative enzymes and contents of accumulated lipofuscin were measured as indicators of oxidative stress. Hepatic MDA and carbonyl contents tended to decrease in 10% buchu diet group compared with control group. Dietary buchu significantly suppressed lipid and protein oxidation in the skin of rats (p<0.05). Contents of hepatic hydroxyl radicals, which exert the highest toxicity among the reactive oxygen species, were significantly decreased in rats fed 10% buchu diet (P<0.05). Activities of antioxidative enzyme, such as superoxide dismutase, catalase, and glutathione peroxidase, tended to increase in liver and skin of rats fed 10% buchu diet, while hepatic catalase activity was significantly increased in buchu group compared with control group. Buchu supplementation significantly inhibited the accumulation of lipofuscin, an end-product of lipid peroxidation reactions induced by reactive oxygen radicals, in eye tissues compared with control diet (p<0.001). In conclusion, buchu supplementation diminished the oxidative stress, so dietary buchu could help to attenuate diabetes complications.

The Effect of Fungicide Carbendazim on Hepatic detoxication systems of rat (살균제 carbendazim이 랫드 간 해독체계에 미치는 영향)

  • Lee, Je-Bong;Shin, Jin-Sup;Jeong, Mi-Hye;Park, Yeon-Ki;Kang, Kyu-Young
    • The Korean Journal of Pesticide Science
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    • v.9 no.4
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    • pp.338-346
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    • 2005
  • Serum alanine aminotransferase(ALT), aspartate aminotransferase (AST), hepatic glutathione, glutathione S-transferase(GST), cytochrome P450 and cytochrome P450 reductase activity were measured to investigate the effects of hepatic detoxication system and metabolic activities of carbendazim in Sprague Dawley(S.D.) male rat at dose levels of 375, 750 or 1,500 mg/kg body weight. Serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST) activities were slightly increased in all test groups after 120 minutes of administration. Glutathione was increased about 20% at high and medium dose level within 120 minutes after administration, while activity of glutathione S-transferase was decreased $36{\sim}50%$. However, the enzyme activity was recovered from all test groups after 240 minutes of administration. Cytochrome P450 and activity of cytochrome P450 reductase were decreased $25{\sim}50%$ until 120 minutes after administration, but recovered after 240 minutes.

Effect of Dietary Protein and Fiber on Ethanol-induced Hepatotoxicity in Rats (흰쥐의 에탄올성 간장해에 미치는 식이 단백질과 섬유소의 영향)

  • 조수열;박은미;이미경;장주연;김명주
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.4
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    • pp.675-681
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    • 1997
  • This study was conducted to investigate the effect of dietary protein and fiber levels on the activities of ethanol metabolizing enzymes of liver in ethanol-treated rats. Sprague-Dawley male rats were fed on diets containing two levels of protein(7, 20%/kg diet) and pectin(5, 10%/kg diet). In ethanol experiments, ethanol(25% v/v) was administered by oral intubation(5g/kg body weight) at the same time once a day Control animals received an isocaloric dose of sucrose. The rats were sacrificed after 5 weeks of feeding periods. Alcohol dehydrogenase and microsomal ethanol oxidizing system activities of hepatic tissue were increased more in ethanol-treated groups than in control groups. Increment of activities predominated in normal protein normal fiber group. Aldehyde dehydrogenase activity was decreased in ethanol-treated groups and significantly decreased in normal Protein normal fiber group. Cytochrome P-450 content was significantly increased in ethanol-treated groups and Predominated in normal protein groups. Xanthine oxidase activity was increased in ethanol-treated groups, but not significantly except normal protein normal fiber group. Glutathione content tended to increase in proportion to level of dietary protein and was higher in normal fiber groups than in high fiber groups, whereas it was decreased by ethanol treatment. Lipid Peroxide content was significantly increased in low Protein normal fiber groups.

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Anti-Obesity Effect of By-Product from Soybean on Mouse Fed a High Fat Diet (고지방 식이로 유도된 비만 마우스에서 대두 부산물인 순물과 침지수의 항비만 효과)

  • Park, Young Mi;Lim, Jae Hwan;Seo, Eul Won
    • Korean Journal of Plant Resources
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    • v.28 no.2
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    • pp.168-177
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    • 2015
  • Here we study the anti-obesity effects of by-product from soybean on mouse fed high fat diet. The body weight gain, visceral and subcutaneous adipose tissue weight, liver and epididymal adipose tissue weight in freeze-dried soybean-soaking-water (SSW) powder fed group showed lower level than those in high fat diet (HFD) group by determining with weight measuring and histological methods. Also, histological analyses of the liver and fat tissues of SSW grouped mice revealed significantly less number of lipid droplets formation and smaller size of adipocytes compared to the HFD group. Moreover, the levels of total serum cholesterol, LDL-cholesterol and the atherogenic index were decreased in the SSW groups. Especially, in SSW group, the levels of phosphorylation of two lipid oxidation enzymes, adenosine monophosphate-activated protein kinase (AMPK) and acetyl-CoA carboxylasse (ACC) were elevated hence that may activate fatty acid oxidation. But AST and ALT levels were not changed in blood. By micro-CT analysis of abdomen, SSW groups significantly showed a tendency to decrease visceral and subcutaneous fats as well as fat-deposited areas compared to HFD group. Taken together, we suggest that soybean soaking water has a function in ameliorating obesity through inhibiting lipid synthesis as well as stimulating fatty acid oxidation.

Biochemical Adaptation of the Oriental Tobacco Budworm, Helicoverpa assulta, to Host-plant Defensive Compounds (기주식물 방어물질에 대한 담배나방의 생화학적 적응)

  • Ahn, Seung-Joon
    • Korean journal of applied entomology
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    • v.61 no.1
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    • pp.143-154
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    • 2022
  • Plant secondary metabolites play an important role in insect-plant interactions. Herbivorous insects have various strategies to cope with the plant defensive compounds. Polyphagous insects feed on a wide variety of plant species, and their detoxification mechanisms are more complex since they tend to respond to a large array of different plant-derived chemicals. Alternatively, oligophagous insects specialize on only a few related plant species and may be expected to have a more efficient form of adaptation. This adaptation could involve either the production of large quantities of enzymes to detoxify their defensive compounds or the sequestration of the compounds or their metabolites. The oriental tobacco budworm, Helicoverpa assulta, is a specialist herbivore, feeding on a few plants of Solanaceae, such as tobacco and hot pepper. Understanding its host-plant adaptation not provides an important insight on physiology, ecology and evolution of specialist herbivores, but also gives a clue to develop management strategies of the pest species such as H. assulta. This paper briefly reviews the specialist, H. assulta, focusing on its host range, larval associations with the host plants, and detoxification mechanisms to nicotine and capsaicin, two characteristic defensive compounds derived from its two major host plants, tobacco and hot pepper, respectively. It summarizes the relevant research over the last half century and provides a future perspective on this subject.

Oxya chinensis sinuosa Mishchenko (Grasshopper) Extract Protects INS-1 Pancreatic β cells against Glucotoxicity-induced Apoptosis and Oxidative Stress (INS-1 췌장 베타 세포에서 벼메뚜기(Oxya chinensis sinuosa Mistshenk) 추출물의 당독성 개선 효과)

  • Park, Jae Eun;Han, Ji Sook
    • Journal of Life Science
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    • v.31 no.11
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    • pp.969-979
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    • 2021
  • Type 2 diabetes is a serious chronic metabolic disease, and the goal of diabetes treatment is to keep blood glucose at a normal level and prevent complications from diabetes. Hyperglycemia is a key pathologic feature of type 2 diabetes that mainly results from insulin resistance and pancreatic β-cell dysfunction. Chronic exposure of β-cells to elevated glucose concentrations induces glucotoxicity. In this study, we examined whether an 80% ethanol extract of Oxya chinensis sinuosa Mishchenko (OEE) protected INS-1 pancreatic β-cells against glucotoxicity-induced apoptosis and oxidative stress. Pretreatment with a high concentration of glucose (high glucose = 30 mM) induced glucotoxicity and apoptosis of INS-1 pancreatic β cells. Treatment with OEE significantly increased cell viability. Treatment with 0.01-0.20 mg/ml OEE dose dependently decreased intracellular reactive oxygen species, lipid peroxidation, and nitric oxide levels and increased insulin secretion in high glucose-pretreated INS-1 β cells. OEE also significantly increased the activities of antioxidant enzymes in response to high-glucose-induced oxidative stress. Moreover, OEE treatment significantly reduced the expressions of pro-apoptotic proteins, including Bax, cytochrome C, caspase-3, and caspase-9, and increased anti-apoptotic Bcl-2 expression. Apoptotic cells were identified using Annexin-V/propidium iodide staining, which revealed that treatment with OEE significantly reduced high-glucose-induced apoptosis. These findings implicate OEE as a valuable functional food in protecting pancreatic β-cells against glucotoxicity-induced apoptosis and oxidative stress.

Curcumin represses lipid accumulation through inhibiting ERK1/2-PPAR-γ signaling pathway and triggering apoptosis in porcine subcutaneous preadipocytes

  • Pan, Shifeng;Chen, Yongfang;Zhang, Lin;Liu, Zhuang;Xu, Xingyu;Xing, Hua
    • Animal Bioscience
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    • v.35 no.5
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    • pp.763-777
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    • 2022
  • Objective: Excessive lipid accumulation in adipocytes results in prevalence of obesity and metabolic syndrome. Curcumin (CUR), a naturally phenolic active ingredient, has been shown to have lipid-lowering effects. However, its underlying mechanisms have remained largely unknown. Therefore, the study aims to determine the effect of CUR on cellular lipid accumulation in porcine subcutaneous preadipocytes (PSPA) and to clarify novel mechanisms. Methods: The PSPA were cultured and treated with or without CUR. Both cell counting Kit-8 and lactate dehydrogenase release assays were used to examine cytotoxicity. Intracellular lipid contents were measured by oil-red-o staining extraction and triglyceride quantification. Apoptosis was determined by flow cytometry and the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labelling assay. Adipogenic and apoptosis genes were analyzed by quantitative polymerase chain reaction and Western blot. Results: The CUR dose-dependently reduced the proliferation and lipid accumulation of PSPA. Noncytotoxic doses of CUR (10 to 20 μM) significantly inhibited extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation and expression of adipogenic genes peroxisome proliferation-activity receptor-γ (PPAR-γ), CCAAT/enhancer binding protein-α, sterol regulatory element-binding protein-1c, adipocyte protein-2, glucose transporter-4 as well as key lipogenic enzymes fatty acid synthase and acetyl-CoA carboxylase, while ERK1/2 activation significantly reversed CUR-reduced lipid accumulation by increasing PPAR-γ. Furthermore, compared with differentiation induced media treated cells, higher dose of CUR (30 μM) significantly decreased the expression of AKT and B-cell lymphoma-2 (BCL-2), while increased the expression of BCL-2-associated X (BAX) and the BAX/BCL-2 expression ratio, suggesting triggered apoptosis by inactivating AKT and increasing BAX/BCL-2 ratio and Caspase-3 expression. Moreover, AKT activation significantly rescued CUR inhibiting lipid accumulation via repressing apoptosis. Conclusion: These results demonstrate that CUR is capable of suppressing differentiation by inhibiting ERK1/2-PPAR-γ signaling pathway and triggering apoptosis via decreasing AKT and subsequently increasing BAX/BCL-2 ratio and Caspase-3, suggesting that CUR provides an important method for the reduction of porcine body fat, as well as the prevention and treatment of human obesity.

Comparative metabolomic analysis in horses and functional analysis of branched chain (alpha) keto acid dehydrogenase complex in equine myoblasts under exercise stress

  • Jeong-Woong, Park;Kyoung Hwan, Kim;Sujung, Kim;Jae-rung, So;Byung-Wook, Cho;Ki-Duk, Song
    • Journal of Animal Science and Technology
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    • v.64 no.4
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    • pp.800-811
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    • 2022
  • The integration of metabolomics and transcriptomics may elucidate the correlation between the genotypic and phenotypic patterns in organisms. In equine physiology, various metabolite levels vary during exercise, which may be correlated with a modified gene expression pattern of related genes. Integrated metabolomic and transcriptomic studies in horses have not been conducted to date. The objective of this study was to detect the effect of moderate exercise on the metabolomic and transcriptomic levels in horses. In this study, using nuclear magnetic resonance (NMR) spectroscopy, we analyzed the concentrations of metabolites in muscle and plasma; we also determined the gene expression patterns of branched chain (alpha) keto acid dehydrogenase kinase complex (BCKDK), which encodes the key regulatory enzymes in branched-chain amino acid (BCAA) catabolism, in two breeds of horses, Thoroughbred and Jeju, at different time intervals. The concentrations of metabolites in muscle and plasma were measured by 1H NMR (nuclear magnetic resonance) spectroscopy, and the relative metabolite levels before and after exercise in the two samples were compared. Subsequently, multivariate data analysis based on the metabolic profiles was performed using orthogonal partial least square discriminant analysis (OPLS-DA), and variable important plots and t-test were used for basic statistical analysis. The stress-induced expression patterns of BCKDK genes in horse muscle-derived cells were examined using quantitative reverse transcription polymerase chain reaction (qPCR) to gain insight into the role of transcript in response to exercise stress. In this study, we found higher concentrations of aspartate, leucine, isoleucine, and lysine in the skeletal muscle of Jeju horses than in Thoroughbred horses. In plasma, compared with Jeju horses, Thoroughbred horses had higher levels of alanine and methionine before exercise; whereas post-exercise, lysine levels were increased. Gene expression analysis revealed a decreased expression level of BCKDK in the post-exercise period in Thoroughbred horses.

A Genetically Encoded Biosensor for the Detection of Levulinic Acid

  • Tae Hyun Kim;Seung-Gyun Woo;Seong Keun Kim;Byeong Hyeon Yoo;Jonghyeok Shin;Eugene Rha;Soo Jung Kim;Kil Koang Kwon;Hyewon Lee;Haseong Kim;Hee-Taek Kim;Bong-Hyun Sung;Seung-Goo Lee;Dae-Hee Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.4
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    • pp.552-558
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    • 2023
  • Levulinic acid (LA) is a valuable chemical used in fuel additives, fragrances, and polymers. In this study, we proposed possible biosynthetic pathways for LA production from lignin and poly(ethylene terephthalate). We also created a genetically encoded biosensor responsive to LA, which can be used for screening and evolving the LA biosynthesis pathway genes, by employing an LvaR transcriptional regulator of Pseudomonas putida KT2440 to express a fluorescent reporter gene. The LvaR regulator senses LA as a cognate ligand. The LA biosensor was first examined in an Escherichia coli strain and was found to be non-functional. When the host of the LA biosensor was switched from E. coli to P. putida KT2440, the LA biosensor showed a linear correlation between fluorescence intensity and LA concentration in the range of 0.156-10 mM LA. In addition, we determined that 0.156 mM LA was the limit of LA detection in P. putida KT2440 harboring an LA-responsive biosensor. The maximal fluorescence increase was 12.3-fold in the presence of 10 mM LA compared to that in the absence of LA. The individual cell responses to LA concentrations reflected the population-averaged responses, which enabled high-throughput screening of enzymes and metabolic pathways involved in LA biosynthesis and sustainable production of LA in engineered microbes.