• Title/Summary/Keyword: Metabolic profiling

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Fabrication of a Partial Genome Microarray of the Methylotrophic Yeast Hansenula polymorpha: Optimization and Evaluation of Transcript Profiling

  • OH , KWAN-SEOK;KWON, OH-SUK;OH, YUN-WI;SOHN, MIN-JEONG;JUNG, SOON-GEE;KIM, YONG-KYUNG;KIM, MIN-GON;RHEE, SANG-KI;GERD GELLISSEN,;KANG, HYUN-AH
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1239-1248
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    • 2004
  • The methylotrophic yeast Hansenula polymorpha has been extensively studied as a model organism for methanol metabolism and peroxisome biogenesis. Recently, this yeast has also attracted attention as a promising host organism for recombinant protein production. Here, we describe the fabrication and evaluation of a DNA chip spotted with 382 open reading frames (ORFs) of H. polymorpha. Each ORF was PCR-amplified using gene-specific primer sets, of which the forward primers had 5'-aminolink. The PCR products were printed in duplicate onto the aldehyde-coated slide glasses to link only the coding strands to the surface of the slide via covalent coupling between amine and aldehyde groups. With the partial genome DNA chip, we compared efficiency of direct and indirect cDNA target labeling methods, and found that the indirect method, using fluorescent-labeled dendrimers, generated a higher hybridization signal-to-noise ratio than the direct method, using cDNA targets labeled by incorporation of fluorescence-labeled nucIeotides during reverse transcription. In addition, to assess the quality of this DNA chip, we analyzed the expression profiles of H. polymorpha cells grown on different carbon sources, such as glucose and methanol, and also those of cells treated with the superoxide­generating drug, menadione. The profiles obtained showed a high-level induction of a set of ORFs involved in methanol metabolism and oxidative stress response in the presence of methanol and menadione, respectively. The results demonstrate the sensitivity and reliability of our arrays to analyze global gene expression changes of H. polymorpha under defined environmental conditions.

Dynamic Gene Expression Profiling of Escherichia coli in Carbon Source Transition from Glucose to Acetate

  • Oh Min-Kyu;Cha Mee-Jeong;Lee Sun-Gu;Rohlin Lars;Liao James C.
    • Journal of Microbiology and Biotechnology
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    • v.16 no.4
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    • pp.543-549
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    • 2006
  • DNA microarray was used to study the transcription profiling of Escherichia coli adapting to acetate as a sole carbon source. Bacteria grown in glucose minimal media were used as a reference. The dynamic expression levels of 3,497 genes were monitored at seven time points during this adaptation. Among the central metabolic genes, the glycolytic and glucose phosphotransferase genes were repressed as the bacteria entered stationary phase, whereas the glyoxylate pathway, TCA cycle, and gluconeogenic genes were induced. Distinct induction or repression patterns were recognized among different pathway genes. For example, the repression of glycolytic genes and the induction of gluconeogenic ones started immediately after glucose was depleted. On the other hand, the regulation of the pentose phosphate pathway genes and glyoxylate genes gradually responded to the glucose depletion or was more related to growth in acetate. When the whole genome was considered, many of the CRP, FadR, and Cra regulons were immediately responsive to the glucose depletion, whereas the $\sigma^s$, Lrp, and IHF regulons were gradually responsive to the glucose depletion. The expression profiling also provided differential regulations between isoenzymes; for example, malic enzymes A (sfcA) and B (maeB). The expression profiles of three genes were confirmed with RT-PCR.

Prospects for Plant Biotechnology and Bioindustry in the 21st Century: Paradigm Shift Driven by Genomics (21세기 식물생명공학과 생물산업의 전망 : 유전체 연구에 의한 Paradigm Shift)

  • LIU Jang Ryol;CHOI Dong-Woog;CHUNG Hwa-Jee
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2002.04a
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    • pp.19-25
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    • 2002
  • Biotechnology in the 21st century will be driven by three emerging technologies: genomics, high-throughput biology, and bioinformatics. These technologies are complementary to one another. A large number of economically important crops are currently subjected to whole genome sequencing. Functional genomics for determining the functions of the genes comprising the given plant genome is under progress by using various means including phenotyping data from transgenic mutants, gene expression profiling data from DNA microarrays, and metabolic profiling data from LC/mass analysis. The aim of plant molecular breeding is shifting from introducing agronomic traits such as herbicide and insect resistance to introducing quality traits such as healthful oils and proteins, which will lead to improved and nutritional food and feed products. Plant molecular breeding is also expected to aim to develop crops for producing human therapeutic and industrial proteins.

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Prospects for Plant Biotechnology and Bioindustry in the 21st Century: Paradigm Shift Driven by Genomics (21세기 식물생명공학과 생물산업의 전망: 유전체 연구에 의한 Paradigm Shift)

  • Liu, Jang-Ryol;Choi, Dong-Woog;Chung, Hwa-Jee
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2002.04b
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    • pp.19-25
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    • 2002
  • Biotechnology in the 21st century will be driven by three emerging technologies: genomics, high-throughput biology, and bioinformatics. These technologies are complementary to one another. A large number of economically important crops are currently subjected to whole genome sequencing. Functional genomics for determining the functions of the genes comprising the given plant genome is under progress by using various means including phenotyping data from transgenic mutants, gene expression profiling data from DNA microarrays, and metabolic profiling data from LC/mass analysis. The aim of plant molecular breeding is shifting from introducing agronomic traits such as herbicide and insect resistance to introducing quality traits such as healthful oils and proteins, which will lead to improved and nutritional food and feed products. Plant molecular breeding is also expected to aim to develop crops for producing human therapeutic and industrial proteins.

  • PDF

Prospects for Plant Biotechnology and Bioindustry in the 21s1 Century: Paradigm Shift Driven by Genomics (21세기 식물생명공학과 생물산업의 전망 : 유전체 연구에 의한 Paradigm Shift)

  • Liu, Jang-Ryol;Choi, Dong-Woog;Chung, Hwa-Jee
    • Journal of Plant Biotechnology
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    • v.29 no.3
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    • pp.145-150
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    • 2002
  • Biotechnology in the 21st century will be driven by three emerging technologies: genomics, high-throughput biology, and bioinformatics. These technologies are complementary to one another. A large number of economically important crops are currently subjected to whole genome sequencing. Functional genomics for determining the functions of the genes comprising the given plant genome is under progress by using various means including phenotyping data from transgenic mutants, gene expression profiling data from DNA microarrays, and metabolic profiling data from LC/mass analysis. The aim of plant molecular breeding is shifting from introducing agronomic traits such as herbicide and insect resistance to introducing quality traits such as healthful oils and proteins, which will lead to improved and nutritional food and feed products. Plant molecular breeding is also expected to aim to develop crops for producing human therapeutic and industrial proteins.

Hypoxia suffocates histone demethylases to change gene expression: a metabolic control of histone methylation

  • Park, Hyunsung
    • BMB Reports
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    • v.50 no.11
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    • pp.537-538
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    • 2017
  • Hypoxia affects various physiological and pathophyological processes. Hypoxia changes the expression of hypoxia-responsive genes through two main pathways. First, hypoxia activates transcription factors (TF) such as Hypoxia-inducible Factor (HIF). Second, hypoxia decreases the activity of Jumonji C domain-containing histone demethylases (JMJDs) that require $O_2$ and ${\alpha}$-Ketoglutarate (${\alpha}$-KG) as substrates. The JMJDs affect gene expression through their regulation of active or repressive histone methylations. Profiling of H3K4me3, H3K9me3, and H3K27me3 under both normoxia and hypoxia identified 75 TFs whose binding motifs were significantly enriched in the methylated regions of the genes. TFs showing similar binding strengths to their target genes might be under the 'metabolic control' which changes histone methylation and gene expression by instant changing catalytic activities of resident histone demethylases.

Serum branch chain amino acids and aromatic amino acids ratio and metabolic risks in Koreans with normal-weight or obesity: a cross-sectional study

  • Ji-Sook Park;Kainat Ahmed;Jung-Eun Yim
    • Korean Journal of Community Nutrition
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    • v.29 no.3
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    • pp.212-221
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    • 2024
  • Objectives: Metabolic disease is strongly associated with future insulin resistance, and its prevalence is increasing worldwide. Thus, identifying early biomarkers of metabolic-related disease based on serum profiling is useful to control future metabolic disease. Our study aimed to assess the association of serum branched chain amino acids (BCAAs) and aromatic amino acids (AAAs) ratio and metabolic disease according to body mass index (BMI) status among Korean adults. Methods: This cross-sectional study included 78 adults aged 20-59 years in Korea. We compared serum amino acid (AA) levels between adults with normal-weight and adults with obesity and investigated biomarkers of metabolic disease. We examined serum AA levels, blood profile, and body composition. We also evaluated the association between serum AAs and metabolic-related disease. Results: The height, weight, BMI, waist circumference, hip circumference, waist-hip-ratio, body fat mass, body fat percent, skeletal muscle mass, systolic blood pressure, and diastolic blood pressure were higher in the group with obesity compared to normal weight group. The group with obesity showed significantly higher levels of BCAA, AAA, and BCAA and AAA ratio. Further, BCAA and AAA ratio were significantly positively correlated with triglyceride, body weight, and skeletal muscle mass. The evaluation of metabolic disease risks revealed an association between the ratios of BCAAs and AAAs, hypertension, and metabolic syndrome. Conclusions: Our study is showed the associations between BCAA and AAA ratio, obesity, and obesity-related diseases using various analytical approaches. The elevated BCAA and AAA ratio could be early biomarkers for predicting future metabolic diseases in Korean population.

1H NMR-based metabolite profiling of diet-induced obesity in a mouse mode

  • Jung, Jee-Youn;Kim, Il-Yong;Kim, Yo-Na;Kim, Jin-Sup;Shin, Jae-Hoon;Jang, Zi-Hey;Lee, Ho-Sub;Hwang, Geum-Sook;Seong, Je-Kyung
    • BMB Reports
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    • v.45 no.7
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    • pp.419-424
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    • 2012
  • High-fat diets (HFD) and high-carbohydrate diets (HCD)-induced obesity through different pathways, but the metabolic differences between these diets are not fully understood. Therefore, we applied proton nuclear magnetic resonance ($^1H$ NMR)-based metabolomics to compare the metabolic patterns between C57BL/6 mice fed HCD and those fed HFD. Principal component analysis derived from $^1H$ NMR spectra of urine showed a clear separation between the HCD and HFD groups. Based on the changes in urinary metabolites, the slow rate of weight gain in mice fed the HCD related to activation of the tricarboxylic acid cycle (resulting in increased levels of citrate and succinate in HCD mice), while the HFD affected nicotinamide metabolism (increased levels of 1-methylnicotineamide, nicotinamide-N-oxide in HFD mice), which leads to systemic oxidative stress. In addition, perturbation of gut microflora metabolism was also related to different metabolic patterns of those two diets. These findings demonstrate that $^1H$ NMR-based metabolomics can identify diet-dependent perturbations in biological pathways.