Investigation of the TCA cycle and the glyoxylate shunt in Escherichia coli BL21 and JM109 using (13)C-NMR/MS.

Abstract:

:Acetate accumulation is a common problem observed in aerobic high cell density Escherichia coli cultures. A previous report has hypothesized that the glyoxylate shunt is active in a low acetate producer, E. coli BL21, and inactive in a high acetate producer, JM109. To further investigate this hypothesis, we now develop a model for the incorporation of (13)C from uniformly labeled glucose into key TCA cycle intermediates. The (13)C isotopomer distributions of oxaloacetate and acetyl-CoA are first determined using NMR and MS techniques. These distributions are next validated by predicting the NMR spectrum of glutamate. Under steady state isotopic conditions, and with knowledge of the full isotopomer distributions of oxaloacetate and acetyl-CoA, the flux ratios through the TCA cycle and the glycoxylate shunt are obtained with respect to the flux through the PPC anaplerotic shunt. We conclude that in BL21, the glyoxylate shunt is active at 22% of the flux through the TCA cycle, and is inactive in JM109. Further, in BL21, the flux through the TCA cycle equals the flux through the PPC shunt, while in JM109 the TCA cycle flux is only third of the flux through the PPC shunt.

journal_name

Biotechnol Bioeng

authors

Noronha SB,Yeh HJ,Spande TF,Shiloach J

subject

Has Abstract

pub_date

2000-05-05 00:00:00

pages

316-27

issue

3

eissn

0006-3592

issn

1097-0290

pii

10.1002/(SICI)1097-0290(20000505)68:3<316::AID-BIT

journal_volume

68

pub_type

杂志文章
  • Achieving complete nitrogen removal by coupling nitritation-anammox and methane-dependent denitrification: A model-based study.

    abstract::The discovery of denitrifying anaerobic methane oxidation (DAMO) processes enables the complete nitrogen removal from wastewater by utilizing the methane produced on site from anaerobic digesters. This model-based study investigated the mechanisms and operational window for efficient nitrogen removal by coupling nitri...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.25866

    authors: Chen X,Guo J,Xie GJ,Yuan Z,Ni BJ

    更新日期:2016-05-01 00:00:00

  • Zonal rate model for axial and radial flow membrane chromatography, part II: model-based scale-up.

    abstract::Membrane chromatography (MC) systems are finding increasing use in downstream processing trains for therapeutic proteins due to the unique mass-transfer characteristics they provide. As a result, there is increased need for model-based methods to scale-up MC units using data collected on a scaled-down unit. Here, a st...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.25217

    authors: Ghosh P,Lin M,Vogel JH,Choy D,Haynes C,von Lieres E

    更新日期:2014-08-01 00:00:00

  • An approach for the stable immobilization of proteins.

    abstract::An approach is presented for the stable covalent immobilization of proteins with a high retention of biological activity. First, chemical modification studies were used to establish enzyme structural and functional properties relevant to the covalent immobilization of an enzyme to agarose based supports. Heparinase wa...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260370305

    authors: Leckband D,Langer R

    更新日期:1991-02-05 00:00:00

  • Simulation of growth and detachment in biofilm systems under defined hydrodynamic conditions.

    abstract::Detachment from biofilms was evaluated using a mixed culture biofilm grown on primary wastewater in a tube reactor. The growth of biofilms and the detachment of biomass from biofilms are strongly influenced by hydrodynamic conditions. In a long-term study, three biofilms were cultivated in a biofilm tube reactor. The ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.10503

    authors: Horn H,Reiff H,Morgenroth E

    更新日期:2003-03-05 00:00:00

  • Impact of external calcium and calcium sensors on ginsenoside Rb1 biosynthesis by Panax notoginseng cells.

    abstract::The effects of external calcium concentrations on biosynthesis of ginsenoside Rb1 and several calcium signal sensors were quantitatively investigated in suspension cultures of Panax notoginseng cells. It was observed that the synthesis of intracellular ginsenoside Rb1 in 3-day incubation was dependent on the medium Ca...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.20386

    authors: Yue CJ,Zhong JJ

    更新日期:2005-02-20 00:00:00

  • Biocatalytic reduction of short-chain carboxylic acids into their corresponding alcohols with syngas fermentation.

    abstract::Short-chain carboxylic acids generated by various mixed- or pure-culture fermentation processes have been considered valuable precursors for production of bioalcohols. While conversion of carboxylic acids into alcohols is routinely performed with catalytic hydrogenation or with strong chemical reducing agents, here, a...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.24786

    authors: Perez JM,Richter H,Loftus SE,Angenent LT

    更新日期:2013-04-01 00:00:00

  • Enhanced acetone-butanol fermentation using repeated fed-batch operation coupled with cell recycle by membrane and simultaneous removal of inhibitory products by adsorption.

    abstract::A novel acetone-butanol production process was developed which integrates a repeated fed-batch fermentation with continuous product removal and cell recycle. The inhibitory product concentrations of the fermentation by Clostridium acetobutylicum were reduced by the simultaneous extraction process using polyvinylpyridi...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260470405

    authors: Yang X,Tsao GT

    更新日期:1995-08-20 00:00:00

  • Split luciferase complementation assay for the analysis of G protein-coupled receptor ligand response in Saccharomyces cerevisiae.

    abstract::The budding yeast Saccharomyces cerevisiae is equipped with G protein-coupled receptors (GPCR). Because the yeast GPCR signaling mechanism is partly similar to that of the mammalian system, S. cerevisiae can be used for a host of mammalian GPCR expression and ligand-mediated activation assays. However, currently avail...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.26255

    authors: Fukutani Y,Ishii J,Kondo A,Ozawa T,Matsunami H,Yohda M

    更新日期:2017-06-01 00:00:00

  • Mechanized systems for media dispensing, inoculation, and replication of microorganisms.

    abstract::Two instruments were developed to mechanize the handling of anaerobic microorganisms for microbial mutant isolation. The instruments automatically dispense liquid or agar medium to large and small 96-well platesand petridishes. Protocols were developed for inoculating different microorganisms, and calibration curves o...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260280905

    authors: Stieglitz B,Defelice CP

    更新日期:1986-09-01 00:00:00

  • Modeling brewers' yeast flocculation

    abstract::Flocculation of yeast cells occurs during the fermentation of beer. Partway through the fermentation the cells become flocculent and start to form flocs. If the environmental conditions, such as medium composition and fluid velocities in the tank, are optimal, the flocs will grow in size large enough to settle. After ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/(sici)1097-0290(19980205)57:3<330::aid-bit

    authors: van Hamersveld EH,van der Lans RG,Caulet PJ,Luyben KC

    更新日期:1998-02-05 00:00:00

  • Modeling thrombus formation and growth.

    abstract::The paper reviews the state-of-the-art in computational modeling of thrombus formation and growth and related phenomena including platelet margination, activation, adhesion, and embolization. Presently, there is a high degree of empiricism in the modeling of thrombus formation. Based on the experimentally observed phy...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章,评审

    doi:10.1002/bit.26343

    authors: Hosseinzadegan H,Tafti DK

    更新日期:2017-10-01 00:00:00

  • Engineering the pentose phosphate pathway to improve hydrogen yield in recombinant Escherichia coli.

    abstract::Among various routes for the biological hydrogen production, the NAD(P)H-dependent pentose phosphate (PP) pathway is the most efficient for the dark fermentation. Few studies, however, have focused on the glucose-6-phosphate 1-dehydrogenase, encoded by zwf, as a key enzyme activating the PP pathway. Although the gluco...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.23259

    authors: Kim YM,Cho HS,Jung GY,Park JM

    更新日期:2011-12-01 00:00:00

  • Adsorption of cellulase from Trichoderma reesei on cellulose and lignacious residue in wood pretreated by dilute sulfuric acid with explosive decompression.

    abstract::The adsorption of cellulase on cellulose and a lignacious residue was examined by using cellulase from Trichoderma reesei, hardwood pretreated by dilute sulfuric acid under high pressure, and a lignacious residue prepared by a complete enzymatic hydrolysis of the pretreated wood. A significant amount of cellulase was ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260360503

    authors: Ooshima H,Burns DS,Converse AO

    更新日期:1990-08-20 00:00:00

  • Peroxidase-catalyzed asymmetric sulfoxidation in organic solvents versus in water.

    abstract::Peroxidase-catalyzed asymmetric sulfoxidations, while synthetically attractive, suffer from relatively low reaction rates due to poor substrate solubilities in water and from appreciable spontaneous oxidation of substrates (especially aryl alkyl sulfides) with H(2)O(2). In this work, we found that both of these shortc...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/1097-0290(20001105)70:3<353::aid-bit13>3.0

    authors: Dai L,Klibanov AM

    更新日期:2000-11-05 00:00:00

  • Hollow-fiber membrane bioreactors using immobilized E. coli for protein synthesis.

    abstract::Actively growing Escherichia coli C600(pBR322), immobilized within the macroporous matrix of asymmetric-wall hollow-fiber membranes, has been propagated to extremely high densities, typically more than 10(12) cells/mL of accessible void volume, in some regions cells accounting for nearly 100% of the available macrovoi...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260251113

    authors: Inloes DS,Smith WJ,Taylor DP,Cohen SN,Michaels AS,Robertson CR

    更新日期:1983-11-01 00:00:00

  • Improved rapid sampling for in vivo kinetics of intracellular metabolites in Saccharomyces cerevisiae.

    abstract::An integrated approach is used to develop a rapid sampling strategy for the quantitative analysis of in vivo kinetic behavior based on measured concentrations of intracellular metabolites in Saccharomyces cerevisiae. Emphasis is laid on small sample sizes during sampling and analysis. Subsecond residence times are acc...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.10048

    authors: Lange HC,Eman M,van Zuijlen G,Visser D,van Dam JC,Frank J,de Mattos MJ,Heijnen JJ

    更新日期:2001-11-20 00:00:00

  • Growth of Leuconostoc mesenteroides NRRL-B523 in an alkaline medium: suboptimal pH growth inhibition of a lactic acid bacterium.

    abstract::Bacterial profile modification (BPM), a form of tertiary oil recovery, diverts water from the water-flooded high-permeability zone into the oil-bearing low-permeability zone. During field use, exopolymer-producing bacteria plug the high-permeability zone only in the immediate vicinity of the injection point (the near-...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.20315

    authors: Wolf BF,Fogler HS

    更新日期:2005-01-05 00:00:00

  • Identification of manipulated variables for a glycosylation control strategy.

    abstract::N-linked glycan distribution affects important end-use characteristics such as the bioactivity and efficacy of many therapeutic proteins, (including monoclonal antibodies), in vivo. Yet, obtaining desired glycan distributions consistently during batch-to-batch production can be challenging for biopharmaceutical manufa...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.25251

    authors: St Amand MM,Radhakrishnan D,Robinson AS,Ogunnaike BA

    更新日期:2014-10-01 00:00:00

  • Single pass tangential flow filtration to debottleneck downstream processing for therapeutic antibody production.

    abstract::As the therapeutic monoclonal antibody (mAb) market continues to grow, optimizing production processes is becoming more critical in improving efficiencies and reducing cost-of-goods in large-scale production. With the recent trends of increasing cell culture titers from upstream process improvements, downstream capaci...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.24377

    authors: Dizon-Maspat J,Bourret J,D'Agostini A,Li F

    更新日期:2012-04-01 00:00:00

  • Scale-up of Artemisia annua L. hairy root cultures produces complex patterns of terpenoid gene expression.

    abstract::Hairy roots grow quickly, reach high densities, and can produce significant amounts of secondary metabolites, yet their scale-up to bioreactors remains challenging. Artemisia annua produces a rich array of terpenoids, including the sesquiterpene, artemisinin, and transformed roots of this species provide a good model ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.10711

    authors: Souret FF,Kim Y,Wyslouzil BE,Wobbe KK,Weathers PJ

    更新日期:2003-09-20 00:00:00

  • Use of frozen bagged seed inoculum for secondary metabolite and bioconversion processes at the pilot scale.

    abstract::Frozen bagged seed inoculum was prepared, thawed and tested for seven cultures. Thawing techniques were developed and other key influences on thawing rate were quantified; seed bag thawing without a water bath rarely required more than 4 to 5 h and was as short as 0.5 to 1 h for lower fill volume bags. Testing include...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.10323

    authors: Junker B,Seeley A,Lester M,Kovatch M,Schmitt J,Boryscwicz S,Lynch J,Zhang J,Greasham R

    更新日期:2002-09-20 00:00:00

  • Tobacco smoke as inducer for gas phase-controlled transgene expression in mammalian cells and mice.

    abstract::Capitalizing on components evolved to metabolize ethanol in Aspergillus nidulans, we previously designed the first molecular gas-gene expression interface using gaseous acetaldehyde as the major inducer. This fungus-derived acetaldehyde-inducible gene regulation (AIR) system operated perfectly and enabled precise and ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.20482

    authors: Weber W,Spielmann M,Daoud El-Baba M,Keller B,Aubel D,Fussenegger M

    更新日期:2005-06-30 00:00:00

  • The effect of medium composition on the acetone-butanol fermentation in continuous culture.

    abstract::The effects of growing Clostridium acetobutylicum NCIB 8052 in a chemostat under conditions of glucose, NH(4) (+), PO(4) (3-), Mg(2+), and Fe(2+) limitation were examined. It was noted that limitation of any major nutrient resulted in the same fermentation pattern. Conditions where minor nutrient levels were reduced a...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260290315

    authors: McNeil B,Kristiansen B

    更新日期:1987-02-01 00:00:00

  • 3D tumor spheroids as in vitro models to mimic in vivo human solid tumors resistance to therapeutic drugs.

    abstract::Three-dimensional cell culture models, such as spheroids, can be used in the process of the development of new anticancer agents because they are able to closely mimic the main features of human solid tumors, namely their structural organization, cellular layered assembling, hypoxia, and nutrient gradients. These prop...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章,评审

    doi:10.1002/bit.26845

    authors: Nunes AS,Barros AS,Costa EC,Moreira AF,Correia IJ

    更新日期:2019-01-01 00:00:00

  • Real-time detection of 13C NMR labeling kinetics in perfused EMT6 mouse mammary tumor cells and betaHC9 mouse insulinomas.

    abstract::A method was developed for obtaining high signal-to-noise 13C NMR spectra of intracellular compounds in metabolically active cultured cells. The method allows TCA cycle labeling kinetics to be determined in real time without significant oxygen transport limitations. Cells were immobilized on the surface of nonporous m...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.20191

    authors: Mancuso A,Beardsley NJ,Wehrli S,Pickup S,Matschinsky FM,Glickson JD

    更新日期:2004-09-30 00:00:00

  • Non-invasive self-referencing electrochemical sensors for quantifying real-time biofilm analyte flux.

    abstract::Current techniques for characterizing biofilm physiology lack the signal filtering capability required for quantifying signals associated with real time biologically active transport. Though a great deal was learned from previous investigations, no results have been reported on the characterization of in vivo, real ti...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.22128

    authors: McLamore ES,Porterfield DM,Banks MK

    更新日期:2009-02-15 00:00:00

  • A limited LCA of bio-adipic acid: manufacturing the nylon-6,6 precursor adipic acid using the benzoic acid degradation pathway from different feedstocks.

    abstract::A limited life cycle assessment (LCA) was performed on a combined biological and chemical process for the production of adipic acid, which was compared to the traditional petrochemical process. The LCA comprises the biological conversion of the aromatic feedstocks benzoic acid, impure aromatics, toluene, or phenol fro...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.23074

    authors: van Duuren JB,Brehmer B,Mars AE,Eggink G,Dos Santos VA,Sanders JP

    更新日期:2011-06-01 00:00:00

  • Fluorophore-labeled carbohydrate analysis of immunoglobulin fusion proteins: Correlation of oligosaccharide content with in vivo clearance profile.

    abstract::CTLA4 is a membrane receptor on cytotoxic T cells whose interaction with the B7 counterreceptor on B cells is important in alloantigen responses. Soluble recombinant human and murine CTLA4 were produced using either Chinese hamster ovary or NS-0 cell lines. Expression vectors were constructed containing the gene codin...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260460502

    authors: Flesher AR,Marzowski J,Wang WC,Raff HV

    更新日期:1995-06-05 00:00:00

  • Resting cells of recombinant E. coli show high epoxidation yields on energy source and high sensitivity to product inhibition.

    abstract::Metabolically active resting (i.e., nongrowing) bacterial cells have a high potential in cofactor-dependent redox biotransformations. Where growing cells require carbon and energy for biomass production, resting cells can potentially exploit their metabolism more efficiently for redox biocatalysis allowing higher spec...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.24404

    authors: Julsing MK,Kuhn D,Schmid A,Bühler B

    更新日期:2012-05-01 00:00:00

  • Characterizing Escherichia coli DH5alpha growth and metabolism in a complex medium using genome-scale flux analysis.

    abstract::Genome-scale flux analysis of Escherichia coli DH5alpha growth in a complex medium was performed to investigate the relationship between the uptake of various nutrients and their metabolic outcomes. During the exponential growth phase, we observed a sequential consumption order of serine, aspartate and glutamate in th...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.22119

    authors: Selvarasu S,Ow DS,Lee SY,Lee MM,Oh SK,Karimi IA,Lee DY

    更新日期:2009-02-15 00:00:00