• 제목/요약/키워드: metabolic flux analysis

검색결과 64건 처리시간 0.035초

Comparative analysis of the transcriptomes and primary metabolite profiles of adventitious roots of five Panax ginseng cultivars

  • Lee, Yun Sun;Park, Hyun-Seung;Lee, Dong-Kyu;Jayakodi, Murukarthick;Kim, Nam-Hoon;Lee, Sang-Choon;Kundu, Atreyee;Lee, Dong-Yup;Kim, Young Chang;In, Jun Gyo;Kwon, Sung Won;Yang, Tae-Jin
    • Journal of Ginseng Research
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    • 제41권1호
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    • pp.60-68
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    • 2017
  • Background: Various Panax ginseng cultivars exhibit a range of diversity for morphological and physiological traits. However, there are few studies on diversity of metabolic profiles and genetic background to understand the complex metabolic pathway in ginseng. Methods: To understand the complex metabolic pathway and related genes in ginseng, we tried to conduct integrated analysis of primary metabolite profiles and related gene expression using five ginseng cultivars showing different morphology. We investigated primary metabolite profiles via gas chromatography-mass spectrometry (GC-MS) and analyzed transcriptomes by Illumina sequencing using adventitious roots grown under the same conditions to elucidate the differences in metabolism underlying such genetic diversity. Results: GC-MS analysis revealed that primary metabolite profiling allowed us to classify the five cultivars into three independent groups and the grouping was also explained by eight major primary metabolites as biomarkers. We selected three cultivars (Chunpoong, Cheongsun, and Sunhyang) to represent each group and analyzed their transcriptomes. We inspected 100 unigenes involved in seven primary metabolite biosynthesis pathways and found that 21 unigenes encoding 15 enzymes were differentially expressed among the three cultivars. Integrated analysis of transcriptomes and metabolomes revealed that the ginseng cultivars differ in primary metabolites as well as in the putative genes involved in the complex process of primary metabolic pathways. Conclusion: Our data derived from this integrated analysis provide insights into the underlying complexity of genes and metabolites that co-regulate flux through these pathways in ginseng.

Periodic Change in DO Concentration for Efficient Poly-${\beta}$-hydroxy-butyrate Production Using Temperature-inducible Recombinant Escherichia coli with Proteome Analysis

  • Abdul Rahman, Nor Aini;Shirai, Yoshihito;Shimizu, Kazuyuki;Hassan, Mohd Ali
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제7권5호
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    • pp.281-288
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    • 2002
  • Recombinant Escherichia coli strain harboring the ${\lambda}$pR-pL promotor and heterologus poly-${\beta}$-hydroxybutyrate (PHB) biosynthesis genes was used to investigate the effect of culture conditions on the efficient PHB production. The expression of phb genes was induced by a temperature upshift from $33^{\circ}C\;to\;38^{\circ}C$. The protein expression levels were measured by using two-dimensional electrophoresis, and the enzyme activities were also measured to understand the effect of culture temperature, carbon sources, and the dissolved oxygen (DO) concentration on the metabolic regulations. AcetylCoA is an important branch point for PHB production. The decrease in DO concentration lowers the citrate synthase activity, thus limit the flux toward the TCA cycle, and increase the flux for PHB production. Since NADPH is required for PHB production, the PHB production does not continue leading the overproduction of acetate and lac-tate. Based on these observations, a new operation was considered where DO concentration was changed periodically, and it was verified its usefulness for the efficient PHB production by experiments.

Succinic Acid Production by Continuous Fermentation Process Using Mannheimia succiniciproducens LPK7

  • Oh, In-Jae;Lee, Hye-Won;Park, Chul-Hwan;Lee, Sang-Yup;Lee, Jin-Won
    • Journal of Microbiology and Biotechnology
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    • 제18권5호
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    • pp.908-912
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    • 2008
  • To achieve a higher succinic acid productivity and evaluate the industrial applicability, this study used Mannheimia succiniciproducens LPK7 (knock-out: ldhA, pflB, pta-ackA), which was recently designed to enhance the productivity of succinic acid and reduce by-product secretion. Anaerobic continuous fermentation of Mannheimia succiniciproducens LPK7 was carried out at different glucose feed concentrations and dilution rates. After extensive fermentation experiments, a succinic acid yield and productivity of 0.38 mol/mol and 1.77 g/l/h, respectively, were achieved with a glucose feed concentration of 18.0 g/l and $0.2\;h^{-1}$ dilution rate. A similar amount of succinic acid production was also produced in batch culture experiments. Therefore, these optimal conditions can be industrially applied for the continuous production of succinic acid. To examine the quantitative balance of the metabolism, a flux distribution analysis was also performed using the metabolic network model of glycolysis and the pentose phosphate pathway.

The Role of Rice Vacuolar Invertase2 in Seed Size Control

  • Lee, Dae-Woo;Lee, Sang-Kyu;Rahman, Md Mizanor;Kim, Yu-Jin;Zhang, Dabing;Jeon, Jong-Seong
    • Molecules and Cells
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    • 제42권10호
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    • pp.711-720
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    • 2019
  • Sink strength optimizes sucrose import, which is fundamental to support developing seed grains and increase crop yields, including those of rice (Oryza sativa). In this regard, little is known about the function of vacuolar invertase (VIN) in controlling sink strength and thereby seed size. Here, in rice we analyzed mutants of two VINs, OsVIN1 and OsVIN2, to examine their role during seed development. In a phenotypic analysis of the T-DNA insertion mutants, only the OsVIN2 mutant osvin2-1 exhibited reduced seed size and grain weight. Scanning electron microscopy analysis revealed that the small seed grains of osvin2-1 can be attributed to a reduction in spikelet size. A significant decrease in VIN activity and hexose level in the osvin2-1 spikelets interfered with spikelet growth. In addition, significant reduction in starch and increase in sucrose, which are characteristic features of reduced turnover and flux of sucrose due to impaired sink strength, were evident in the pre-storage stage of osvin2-1 developing grains. In situ hybridization analysis found that expression of OsVIN2 was predominant in the endocarp of developing grains. A genetically complemented line with a native genomic clone of OsVIN2 rescued reduced VIN activity and seed size. Two additional mutants, osvin2-2 and osvin2-3 generated by the CRISPR/Cas9 method, exhibited phenotypes similar to those of osvin2-1 in spikelet and seed size, VIN activity, and sugar metabolites. These results clearly demonstrate an important role of OsVIN2 as sink strength modulator that is critical for the maintenance of sucrose flux into developing seed grains.

Macromolecular and Elemental Composition Analyses of Leuconostoc mesenteroides ATCC 8293 Cultured in a Chemostat

  • Bang, Jeongsu;Li, Ling;Seong, Hyunbin;Kwon, Ye Won;Jeong, Eun Ji;Lee, Dong-Yup;Han, Nam Soo
    • Journal of Microbiology and Biotechnology
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    • 제27권5호
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    • pp.939-942
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    • 2017
  • The cellular composition and metabolic compounds of Leuconostoc mesenteroides ATCC 8293 were analyzed after cultivation in an anaerobic chemostat. The macromolecular composition was 24.4% polysaccharide, 29.7% protein, 7.9% lipid, 2.9% DNA, and 7.4% RNA. Its amino acid composition included large amounts of lysine, glutamic acid, alanine, and leucine. Elements were in the order of C > O > N > H > S. The metabolites in chemostat culture were lactic acid (73.34 mM), acetic acid (7.69 mM), and mannitol (9.93 mM). These data provide a first view of the cellular composition of L. mesenteroides for use in metabolic flux analysis.

식물의 탄소대사공학 연구동향 (Current status on carbon metabolic engineering in plants)

  • 김동헌;이시명;박종석;김수진;김범기;윤인선;김둘이;변명옥
    • Journal of Plant Biotechnology
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    • 제37권2호
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    • pp.205-211
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    • 2010
  • Yield productivity of staple crops must be increased at least 50% by 2050, in order to feed the world population which is expected to reach 90 billions. Photosynthetic carbon assimilation and carbohydrate metabolism leading to the production of starch would be the final frontier to quest for new sources of technology enabling such a drastic increase of crop productivity. In this review, attempts to genetically engineer plant photosynthetic carbon reduction cycle and metabolic pathways to increase starch production are introduced.

Hyperpolarized Carbon-13 Magnetic Resonance Imaging: Technical Considerations and Clinical Applications

  • Ying-Chieh Lai;Ching-Yi Hsieh;Yu-Hsiang Juan;Kuan-Ying Lu;Hsien-Ju Lee;Shu-Hang Ng;Yung-Liang Wan;Gigin Lin
    • Korean Journal of Radiology
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    • 제25권5호
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    • pp.459-472
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    • 2024
  • Hyperpolarized (HP) carbon-13 (13C) MRI represents an innovative approach for noninvasive, real-time assessment of dynamic metabolic flux, with potential integration into routine clinical MRI. The use of [1-13C]pyruvate as a probe and its conversion to [1-13C]lactate constitute an extensively explored metabolic pathway. This review comprehensively outlines the establishment of HP 13C-MRI, covering multidisciplinary team collaboration, hardware prerequisites, probe preparation, hyperpolarization techniques, imaging acquisition, and data analysis. This article discusses the clinical applications of HP 13C-MRI across various anatomical domains, including the brain, heart, skeletal muscle, breast, liver, kidney, pancreas, and prostate. Each section highlights the specific applications and findings pertinent to these regions, emphasizing the potential versatility of HP 13C-MRI in diverse clinical contexts. This review serves as a comprehensive update, bridging technical aspects with clinical applications and offering insights into the ongoing advancements in HP 13C-MRI.

폐암조직에서 중성자 방사화 분석법을 이용한 미량 원소 분석 (Trace Element Analysis by Neutron Activastion Analysis in the Human Cancer Tissue)

  • 임상무;조재일;심영목;정영주;조승연;정용삼
    • 대한핵의학회지
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    • 제27권1호
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    • pp.104-111
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    • 1993
  • Trace elements are important components in the biological system, as a structural material and metabolic controller. Neutron activation analysis (NAA) with high neutron flux and high energy resolution Ge (Li) detector coupled to multichannel analyzer (MCA) has been one of the most accurate method for the determination of ultra-trace level components, and is applicable to biological material. In human body, the NAA can be used for quantitation of trace elements in various organs and tissue with endocrinological and metabolic disease and industrial metal poisoning. In this study, Triga Mark III nuclear reactor in Korea Atomic Research Institute was used for quantitation of trace eleement in human lung cancer tissues by neutron activation analysis. In the squamous cell carcinoma tissues, Br, Hg, La, Sb, Sc, Cl, Fe and I content were lower than normal lung tissues, and K, Rb and Se content were higher. In the adenocarcinoma tissues, Fe, Au, La, Sc and Zn content were lower than normal lung tissues, and Rb, Co and Se content were higher. Rb content was higher in the adenocarcinoma tissues than in the squamous cell carcinoma tissues. Fe and Na content were higher in the squamous cell carcinoma tissues than in the adenocarcinoma tissues.

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Simulation of Dynamic Behavior of Glucose- and Tryptophan-Grown Escherichia coli Using Constraint-Based Metabolic Models with a Hierarchical Regulatory Network

  • Lee Sung-Gun;Kim Yu-Jin;Han Sang-Il;Oh You-Kwan;Park Sung-Hoon;Kim Young-Han;Hwang Kyu-Suk
    • Journal of Microbiology and Biotechnology
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    • 제16권6호
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    • pp.993-998
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    • 2006
  • We earlier suggested a hierarchical regulatory network using defined modeling symbols and weights in order to improve the flux balance analysis (FBA) with regulatory events that were represented by if-then rules and Boolean logic. In the present study, the simulation results of the models, which were developed and improved from the previou model by incorporating a hierarchical regulatory network into the FBA, were compared with the experimental outcome of an aerobic batch growth of E. coli on glucose and tryptophan. From the experimental result, a diauxic growth curve was observed, reflecting growth resumption, when tryptophan was used as an alternativee after the supply of glucose was exhausted. The model parameters, the initial concentration of substrates (0.92 mM glucose and 1 mM tryptophan), cell density (0.0086 g biomass/1), the maximal uptake rates of substrates (5.4 mmol glucose/g DCW h and 1.32 mmol tryptophan/g DCW h), and lag time (0.32 h) were derived from the experimental data for more accurate prediction. The simulation results agreed with the experimental outcome of the temporal profiles of cell density and glucose, and tryptophan concentrations.

Dynamic Behavior of Regulatory Elements in the Hierarchical Regulatory Network of Various Carbon Sources-Grown Escherichia coli

  • Lee, Sung-Gun;Hwang, Kyu-Suk;Kim, Cheol-Min
    • Journal of Microbiology and Biotechnology
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    • 제15권3호
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    • pp.551-559
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    • 2005
  • The recent rapid increase in genomic data related to many microorganisms and the development of computational tools to accurately analyze large amounts of data have enabled us to design several kinds of simulation approaches for the complex behaviors of cells. Among these approaches, dFBA (dynamic flux balance analysis), which utilizes FBA, differential equations, and regulatory events, has correctly predicted cellular behaviors under given environmental conditions. However, until now, dFBA has centered on substrate concentration, cell growth, and gene on/off, but a detailed hierarchical structure of a regulatory network has not been taken into account. The use of Boolean rules for regulatory events in dFBA has limited the representation of interactions between specific regulatory proteins and genes and the whole transcriptional regulation mechanism with environmental change. In this paper, we adopted the operon as the basic structure, constructed a hierarchical structure for a regulatory network with defined fundamental symbols, and introduced a weight between symbols in order to solve the above problems. Finally, the total control mechanism of regulatory elements (operons, genes, effectors, etc.) with time was simulated through the linkage of dFBA with regulatory network modeling. The lac operon, trp operon, and tna operon in the central metabolic network of E. coli were chosen as the basic models for control patterns. The suggested modeling method in this study can be adopted as a basic framework to describe other transcriptional regulations, and provide biologists and engineers with useful information on transcriptional regulation mechanisms under extracellular environmental change.