Metabolic flux analysis in Synechocystis using isotope distribution from 13C-labeled glucose.

Abstract:

:Using the carbon isotope labeling technique, the response of cyanobacterial central carbon metabolism to the change in environmental conditions was investigated. Synechocystis was grown in the heterotrophic and mixotrophic cultures fed with 13C-labeled glucose. The labeling patterns of the amino acids in biomass hydrolysates for both cultures were detected by the two-dimensional 1H-13C correlation nuclear magnetic resonance (2D 1H-13C COSY NMR) spectroscopy and gas chromatography-mass spectrometry (GC-MS) technique. The in vivo intracellular flux distributions were then quantitated from the labeling measurements and metabolite balances using a parameters fitting approach. From the estimated flux distributions, it was found that the pentose phosphate pathway was the major pathway of glucose catabolism in the heterotrophic culture, while in the mixotrophic culture, the flux of CO2 fixation through the Calvin cycle was about two-fold of the glucose input flux. The relative flux through the phosphoenolpyruvate carboxylase was very high in both cultures, and this reaction represented about 25% of the assimilated CO2 in the mixotrophic culture. More importantly, we found a substantial outflow from the tricarboxylic acid cycle to glycolysis pathway carried by the malic enzyme, demonstrating the operation of a C4 pathway in cyanobacterial cells through the PEP carboxylase and malic enzyme. The estimated flux distributions also revealed that the NADPH synthesis was in excess relative to its requirement, and the excess NADPH might be reoxidized in cyanobacterial respiration to provide the energy for cellular requirement. Moreover, the analyzed result also suggested that the activity of the respiratory electron transport chain in cyanobacterial cells was not inhibited by light.

journal_name

Metab Eng

journal_title

Metabolic engineering

authors

Yang C,Hua Q,Shimizu K

doi

10.1006/mben.2002.0226

subject

Has Abstract

pub_date

2002-07-01 00:00:00

pages

202-16

issue

3

eissn

1096-7176

issn

1096-7184

pii

S1096717602902260

journal_volume

4

pub_type

杂志文章
  • Metabolic engineering of aerobic succinate production systems in Escherichia coli to improve process productivity and achieve the maximum theoretical succinate yield.

    abstract::The potential to produce succinate aerobically in Escherichia coli would offer great advantages over anaerobic fermentation in terms of faster biomass generation, carbon throughput, and product formation. Genetic manipulations were performed on two aerobic succinate production systems to increase their succinate yield...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2004.10.003

    authors: Lin H,Bennett GN,San KY

    更新日期:2005-03-01 00:00:00

  • Ethyl acetate production by the elusive alcohol acetyltransferase from yeast.

    abstract::Ethyl acetate is an industrially relevant ester that is currently produced exclusively through unsustainable processes. Many yeasts are able to produce ethyl acetate, but the main responsible enzyme has remained elusive, hampering the engineering of novel production strains. Here we describe the discovery of a new enz...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2017.03.004

    authors: Kruis AJ,Levisson M,Mars AE,van der Ploeg M,Garcés Daza F,Ellena V,Kengen SWM,van der Oost J,Weusthuis RA

    更新日期:2017-05-01 00:00:00

  • Characterization and engineering of 3-ketosteroid-△1-dehydrogenase and 3-ketosteroid-9α-hydroxylase in Mycobacterium neoaurum ATCC 25795 to produce 9α-hydroxy-4-androstene-3,17-dione through the catabolism of sterols.

    abstract::3-Ketosteroid-△(1)-dehydrogenase (KstD) is a key enzyme involved in the microbial catabolism of sterols. Here, three homologues of KstD were characterized from Mycobacterium neoaurum ATCC 25795, showing distinct substrate preferences and transcriptional responses to steroids. Single deletion of any MN-kstD failed to r...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2014.05.005

    authors: Yao K,Xu LQ,Wang FQ,Wei DZ

    更新日期:2014-07-01 00:00:00

  • A biosynthetic route for polysialylating proteins in Escherichia coli.

    abstract::Polysialic acid (polySia) is a posttranslational modification found on only a handful of proteins in the central nervous and immune systems. The addition of polySia to therapeutic proteins improves pharmacokinetics and reduces immunogenicity. To date, polysialylation of therapeutic proteins has only been achieved in v...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2017.10.012

    authors: Keys TG,Wetter M,Hang I,Rutschmann C,Russo S,Mally M,Steffen M,Zuppiger M,Müller F,Schneider J,Faridmoayer A,Lin CW,Aebi M

    更新日期:2017-11-01 00:00:00

  • Overproduction of free fatty acids in E. coli: implications for biodiesel production.

    abstract::Whereas microbial fermentation processes for producing ethanol and related alcohol biofuels are well established, biodiesel (methyl esters of fatty acids) is exclusively derived from plant oils. Slow cycle times for engineering oilseed metabolism and the excessive accumulation of glycerol as a byproduct are two major ...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2008.08.006

    authors: Lu X,Vora H,Khosla C

    更新日期:2008-11-01 00:00:00

  • A comparative study of global stress gene regulation in response to overexpression of recombinant proteins in Escherichia coli.

    abstract::Global gene regulation throughout the Escherichia coli stress response to overexpression of each of five recombinant proteins was evaluated. Reverse-transcriptase polymerase chain reaction-amplified mRNA from induced and control cells were hybridized with a DNA array of Kohara clones representing 16% (700 genes) of th...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1006/mben.2000.0148

    authors: Gill RT,Valdes JJ,Bentley WE

    更新日期:2000-07-01 00:00:00

  • Computational tools for isotopically instationary 13C labeling experiments under metabolic steady state conditions.

    abstract::(13)C metabolic flux analysis (MFA) has become an important and powerful tool for the quantitative analysis of metabolic networks in the framework of metabolic engineering. Isotopically instationary (13)C MFA under metabolic stationary conditions is a promising refinement of classical stationary MFA. It accounts for t...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2006.05.006

    authors: Nöh K,Wahl A,Wiechert W

    更新日期:2006-11-01 00:00:00

  • Mapping photoautotrophic metabolism with isotopically nonstationary (13)C flux analysis.

    abstract::Understanding in vivo regulation of photoautotrophic metabolism is important for identifying strategies to improve photosynthetic efficiency or re-route carbon fluxes to desirable end products. We have developed an approach to reconstruct comprehensive flux maps of photoautotrophic metabolism by computational analysis...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2011.08.002

    authors: Young JD,Shastri AA,Stephanopoulos G,Morgan JA

    更新日期:2011-11-01 00:00:00

  • 13C metabolic flux analysis at a genome-scale.

    abstract::Metabolic models used in 13C metabolic flux analysis generally include a limited number of reactions primarily from central metabolism. They typically omit degradation pathways, complete cofactor balances, and atom transition contributions for reactions outside central metabolism. This study addresses the impact on pr...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2015.08.006

    authors: Gopalakrishnan S,Maranas CD

    更新日期:2015-11-01 00:00:00

  • Estradiol stimulates the biosynthetic pathways of breast cancer cells: detection by metabolic flux analysis.

    abstract::Selective estrogen receptor (ER) modulators are highly successful breast cancer therapies, but they are not effective in patients with ER negative and selective estrogen receptor modulator (SERM)-resistant tumors. Understanding the mechanisms of estrogen-stimulated proliferation may provide a route to design estrogen-...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2006.06.005

    authors: Forbes NS,Meadows AL,Clark DS,Blanch HW

    更新日期:2006-11-01 00:00:00

  • Streptomyces species: Ideal chassis for natural product discovery and overproduction.

    abstract::There is considerable interest in mining organisms for new natural products (NPs) and in improving methods to overproduce valuable NPs. Because of the rapid development of tools and strategies for metabolic engineering and the markedly increased knowledge of the biosynthetic pathways and genetics of NP-producing organ...

    journal_title:Metabolic engineering

    pub_type: 杂志文章,评审

    doi:10.1016/j.ymben.2018.05.015

    authors: Liu R,Deng Z,Liu T

    更新日期:2018-11-01 00:00:00

  • An evolutionary metabolic engineering approach for enhancing lipogenesis in Yarrowia lipolytica.

    abstract::Lipogenic organisms provide an ideal platform for biodiesel and oleochemical production. Through our previous rational metabolic engineering efforts, lipogenesis titers in Yarrowia lipolytica were significantly enhanced. However, the resulting strain still suffered from decreased biomass generation rates. Here, we emp...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2015.02.003

    authors: Liu L,Pan A,Spofford C,Zhou N,Alper HS

    更新日期:2015-05-01 00:00:00

  • Glutamate excretion as a major kinetic bottleneck for the thermally triggered production of glutamic acid by Corynebacterium glutamicum.

    abstract::The study was aimed at evaluating the extent of flux control exercised by the amino acid excretion step on the glutamate production flux in C. glutamicum 2262 strain that is induced for glutamate excretion by an upward temperature shift. Cells initially induced to excrete glutamate were cultivated at different control...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1006/mben.1999.0129

    authors: Lapujade P,Goergen JL,Engasser JM

    更新日期:1999-07-01 00:00:00

  • Improvement of 1,3-propanediol production using an engineered cyanobacterium, Synechococcus elongatus by optimization of the gene expression level of a synthetic metabolic pathway and production conditions.

    abstract::The introduction of a synthetic metabolic pathway consisting of multiple genes derived from various organisms enables cyanobacteria to directly produce valuable chemicals from carbon dioxide. We previously constructed a synthetic metabolic pathway composed of genes from Escherichia coli, Saccharomyces cerevisiae, and ...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2016.12.001

    authors: Hirokawa Y,Maki Y,Hanai T

    更新日期:2017-01-01 00:00:00

  • Colour bio-factories: Towards scale-up production of anthocyanins in plant cell cultures.

    abstract::Anthocyanins are widely distributed, glycosylated, water-soluble plant pigments, which give many fruits and flowers their red, purple or blue colouration. Their beneficial effects in a dietary context have encouraged increasing use of anthocyanins as natural colourants in the food and cosmetic industries. However, the...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2018.06.004

    authors: Appelhagen I,Wulff-Vester AK,Wendell M,Hvoslef-Eide AK,Russell J,Oertel A,Martens S,Mock HP,Martin C,Matros A

    更新日期:2018-07-01 00:00:00

  • Efficient mining of natural NADH-utilizing dehydrogenases enables systematic cofactor engineering of lysine synthesis pathway of Corynebacterium glutamicum.

    abstract::Increasing the availability of NADPH is commonly used to improve lysine production by Corynebacterium glutamicum since 4 mol of NADPH are required for the synthesis of 1 mol of lysine. Alternatively, engineering of enzymes in lysine synthesis pathway to utilize NADH directly can also be explored for cofactor balance d...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2018.11.006

    authors: Wu W,Zhang Y,Liu D,Chen Z

    更新日期:2019-03-01 00:00:00

  • Colored Petri net modeling and simulation of signal transduction pathways.

    abstract::Presented herein is a methodology for quantitatively analyzing the complex signaling network by resorting to colored Petri nets (CPN). The mathematical as well as Petri net models for two basic reaction types were established, followed by the extension to a large signal transduction system stimulated by epidermal grow...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2005.10.001

    authors: Lee DY,Zimmer R,Lee SY,Park S

    更新日期:2006-03-01 00:00:00

  • A method for analysis and design of metabolism using metabolomics data and kinetic models: Application on lipidomics using a novel kinetic model of sphingolipid metabolism.

    abstract::We present a model-based method, designated Inverse Metabolic Control Analysis (IMCA), which can be used in conjunction with classical Metabolic Control Analysis for the analysis and design of cellular metabolism. We demonstrate the capabilities of the method by first developing a comprehensively curated kinetic model...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2016.04.002

    authors: Savoglidis G,da Silveira Dos Santos AX,Riezman I,Angelino P,Riezman H,Hatzimanikatis V

    更新日期:2016-09-01 00:00:00

  • Iterative algorithm-guided design of massive strain libraries, applied to itaconic acid production in yeast.

    abstract::Metabolic engineering requires multiple rounds of strain construction to evaluate alternative pathways and enzyme concentrations. Optimizing multigene pathways stepwise or by randomly selecting enzymes and expression levels is inefficient. Here, we apply methods from design of experiments (DOE) to guide the constructi...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2018.05.002

    authors: Young EM,Zhao Z,Gielesen BEM,Wu L,Benjamin Gordon D,Roubos JA,Voigt CA

    更新日期:2018-07-01 00:00:00

  • Metabolic changes in a pyruvate kinase gene deletion mutant of Corynebacterium glutamicum ATCC 13032.

    abstract::To investigate primary effects of a pyruvate kinase (PYK) defect on glucose metabolism in Corynebacterium glutamicum, a pyk-deleted mutant was derived from wild-type C. glutamicum ATCC13032 using the double-crossover chromosome replacement technique. The mutant was then evaluated under glutamic acid-producing conditio...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2010.01.004

    authors: Sawada K,Zen-in S,Wada M,Yokota A

    更新日期:2010-07-01 00:00:00

  • Reversal of β-oxidative pathways for the microbial production of chemicals and polymer building blocks.

    abstract::β-Oxidation is the ubiquitous metabolic strategy to break down fatty acids. In the course of this four-step process, two carbon atoms are liberated per cycle from the fatty acid chain in the form of acetyl-CoA. However, typical β-oxidative strategies are not restricted to monocarboxylic (fatty) acid degradation only, ...

    journal_title:Metabolic engineering

    pub_type: 杂志文章,评审

    doi:10.1016/j.ymben.2017.05.004

    authors: Kallscheuer N,Polen T,Bott M,Marienhagen J

    更新日期:2017-07-01 00:00:00

  • Metabolic engineering of Clostridium acetobutylicum for butyric acid production with high butyric acid selectivity.

    abstract::A typical characteristic of the butyric acid-producing Clostridium is coproduction of both butyric and acetic acids. Increasing the butyric acid selectivity important for economical butyric acid production has been rather difficult in clostridia due to their complex metabolic pathways. In this work, Clostridium acetob...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2014.03.004

    authors: Jang YS,Im JA,Choi SY,Lee JI,Lee SY

    更新日期:2014-05-01 00:00:00

  • Metabolic engineering of tomato for high-yield production of astaxanthin.

    abstract::Dietary carotenoids have been shown to be beneficial to health by decreasing the risk of many diseases. Attempts to enhance carotenoids in food crops have been successful although higher plants appear to resist big changes of carotenoid biosynthesis by metabolic engineering. Here we report the generation of a more nut...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2013.02.005

    authors: Huang JC,Zhong YJ,Liu J,Sandmann G,Chen F

    更新日期:2013-05-01 00:00:00

  • Spatial organization of enzymes for metabolic engineering.

    abstract::As synthetic pathways built from exogenous enzymes become more complicated, the probability of encountering undesired interactions with host organisms increases, thereby lowering product titer. An emerging strategy to combat this problem is to spatially organize pathway enzymes into multi-protein complexes, where high...

    journal_title:Metabolic engineering

    pub_type: 杂志文章,评审

    doi:10.1016/j.ymben.2011.09.003

    authors: Lee H,DeLoache WC,Dueber JE

    更新日期:2012-05-01 00:00:00

  • High-level production of valine by expression of the feedback inhibition-insensitive acetohydroxyacid synthase in Saccharomyces cerevisiae.

    abstract::Valine, which is one of the branched-chain amino acids (BCAAs) essential for humans, is widely used in animal feed, dietary supplements and pharmaceuticals. At the commercial level, valine is usually produced by bacterial fermentation from glucose. However, valine biosynthesis can also proceed in the yeast Saccharomyc...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2018.02.011

    authors: Takpho N,Watanabe D,Takagi H

    更新日期:2018-03-01 00:00:00

  • Metabolomics-driven approach to solving a CoA imbalance for improved 1-butanol production in Escherichia coli.

    abstract::High titer 1-butanol production in Escherichia coli has previously been achieved by overexpression of a modified clostridial 1-butanol production pathway and subsequent deletion of native fermentation pathways. This strategy couples growth with production as 1-butanol pathway offers the only available terminal electro...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2017.04.003

    authors: Ohtake T,Pontrelli S,Laviña WA,Liao JC,Putri SP,Fukusaki E

    更新日期:2017-05-01 00:00:00

  • Coupled incremental precursor and co-factor supply improves 3-hydroxypropionic acid production in Saccharomyces cerevisiae.

    abstract::3-Hydroxypropionic acid (3-HP) is an attractive platform chemical, which can be used to produce a variety of commodity chemicals, such as acrylic acid and acrylamide. For enabling a sustainable alternative to petrochemicals as the feedstock for these commercially important chemicals, fermentative production of 3-HP is...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2014.01.005

    authors: Chen Y,Bao J,Kim IK,Siewers V,Nielsen J

    更新日期:2014-03-01 00:00:00

  • Improving the performance of solventogenic clostridia by reinforcing the biotin synthetic pathway.

    abstract::An efficient production process is important for industrial microorganisms. The cellular efficiency of solventogenic clostridia, a group of anaerobes capable of producing a wealth of bulk chemicals and biofuels, must be improved for competitive commercialization. Here, using Clostridium acetobutylicum, a species of so...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2016.02.006

    authors: Yang Y,Lang N,Yang G,Yang S,Jiang W,Gu Y

    更新日期:2016-05-01 00:00:00

  • RCA-I-resistant CHO mutant cells have dysfunctional GnT I and expression of normal GnT I in these mutants enhances sialylation of recombinant erythropoietin.

    abstract::A large number of CHO glycosylation mutants were isolated by Ricinus communis agglutinin-I (RCA-I). Complementation tests revealed that all these mutant lines possessed a dysfunctional N-acetylglucosaminyltransferase I (GnT I) gene. Sequencing analyses on the GnT I cDNAs isolated from 16 mutant lines led to the identi...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2010.03.002

    authors: Goh JS,Zhang P,Chan KF,Lee MM,Lim SF,Song Z

    更新日期:2010-07-01 00:00:00

  • Construction of lycopene-overproducing Saccharomyces cerevisiae by combining directed evolution and metabolic engineering.

    abstract::Improved supply of farnesyl diphosphate (FPP) is often considered as a typical strategy for engineering Saccharomyces cerevisiae towards efficient terpenoid production. However, in the engineered strains with enhanced precursor supply, the production of the target metabolite is often impeded by insufficient capacity o...

    journal_title:Metabolic engineering

    pub_type: 杂志文章

    doi:10.1016/j.ymben.2015.04.009

    authors: Xie W,Lv X,Ye L,Zhou P,Yu H

    更新日期:2015-07-01 00:00:00