Yield, solubility and conformational quality of soluble proteins are not simultaneously favored in recombinant Escherichia coli.


:Many enzymes or fluorescent proteins produced in Escherichia coli are enzymatically active or fluorescent respectively when deposited as inclusion bodies. The occurrence of insoluble but functional protein species with native-like secondary structure indicates that solubility and conformational quality of recombinant proteins are not coincident parameters, and suggests that both properties can be engineered independently. We have here proven this principle by producing elevated yields of a highly fluorescent but insoluble green fluorescent protein (GFP) in a DnaK- background, and further enhancing its solubility through adjusting the growth temperature and GFP gene expression rate. The success of such a two-step approach confirms the independent control of solubility and conformational quality, advocates for new routes towards high quality protein production and intriguingly, proves that high protein yields dramatically compromise the conformational quality of soluble versions.


Biotechnol Bioeng


Martínez-Alonso M,García-Fruitós E,Villaverde A




Has Abstract


2008-12-15 00:00:00












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    abstract::A denitrifying consortium capable of transforming carbon tetrachloride (CCl(4)) was cultured from aquifer sediment from the U.S. Department of Energy's Hanford Site in southeastern Washington State. To understand the kinetics of the biological destruction of CCl(4) by these microbes, a set of experiments, the conditio...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Petersen JN,Skeen RS,Amos KM,Hooker BS

    更新日期:1994-03-15 00:00:00

  • Evaluation of heart tissue viability under redox-magnetohydrodynamics conditions: toward fine-tuning flow in biological microfluidics applications.

    abstract::A microfluidic system containing a chamber for heart tissue biopsies, perfused with Krebs-Henseleit buffer containing glucose and antibiotic (KHGB) using peristaltic pumps and continuously stimulated, was used to evaluate tissue viability under redox-magnetohydrodynamics (redox-MHD) conditions. Redox-MHD possesses uni...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Cheah LT,Fritsch I,Haswell SJ,Greenman J

    更新日期:2012-07-01 00:00:00

  • Kinetic study of sulfide leaching by galvanic interaction between chalcopyrite, pyrite, and sphalerite in the presence of T. ferrooxidans (30 degrees C) and a thermophilic microorganism (55 degrees C).

    abstract::A systematic, kinetic study and comparison of the leaching of mixed metal sulfides by galvanic conversion and in the presence of bacteria has been carried out for the first time using both powder (-100 to -400 mesh) and larger (bulk) specimen systems. The rates of dissolution of copper from chalcopyrite and zinc from ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Mehta AP,Murr LE

    更新日期:1982-04-01 00:00:00

  • Periodical batch culture of the immobilized growing fungi Sporotrichum cellulophilum producing cellulase in the nonwoven materials.

    abstract::The thermophilic fungus Sporotrichum cellulophilum was immobilized with nonwoven materials for cellulase production. The cellulose powder concentration in the medium was an important factor controlling cellulase production. When the cellulose powder concentration in the nonwoven materials was more than 4%, cellulase p...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Tamada M,Kasai N,Kumakura M,Kaetsu I

    更新日期:1986-08-01 00:00:00

  • Distinct timescales of RNA regulators enable the construction of a genetic pulse generator.

    abstract::To build complex genetic networks with predictable behaviors, synthetic biologists use libraries of modular parts that can be characterized in isolation and assembled together to create programmable higher-order functions. Characterization experiments and computational models for gene regulatory parts operating in iso...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Westbrook A,Tang X,Marshall R,Maxwell CS,Chappell J,Agrawal DK,Dunlop MJ,Noireaux V,Beisel CL,Lucks J,Franco E

    更新日期:2019-05-01 00:00:00

  • Isolation of a fluffy mutant of Aspergillus niger from chemostat culture and its potential use as a morphologically stable host for protein production.

    abstract::Chemostat cultivation of Aspergillus niger and other filamentous fungi is often hindered by the spontaneous appearance of morphologic mutants. Using the Variomixing bioreactor and applying different chemostat conditions we tried to optimize morphologic stability in both ammonium- and glucose-limited cultures. In most ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: van de Vondervoort PJ,Poulsen BR,Ruijter GJ,Schuleit T,Visser J,Iversen JJ

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  • Self-renaturing enzymes: design of an enzyme-chaperone chimera as a new approach to enzyme stabilization.

    abstract::Molecular chaperones in aqueous-organic mixtures can broaden the utility of biocatalysis by stabilizing enzymes in denaturing conditions. We have designed a self-renaturing enzyme-chaperone chimera consisting of penicillin amidase and a thermophilic chaperonin that functions in aqueous-organic mixtures. The flexible l...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Bergeron LM,Gomez L,Whitehead TA,Clark DS

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  • Construction, expression, and analysis of recombinant HIV gp41 constructs containing a novel cellular binding domain.

    abstract::The gp41 polypeptide of human immunodeficiency virus (HIV) contains an immunosuppressive domain, an epitope which elicits specific cytolytic T cell responses to HIV, and a complement Clq interactive domain. In addition, a synthetic peptide called CS3, derived from gp41 (amino acids 576-593 of gp160) and contiguous wit...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Kestler DP,Henderson LA,Noti JD

    更新日期:1993-06-05 00:00:00

  • Cell-surface exposure of a hybrid 3-cohesin scaffoldin allowing the functionalization of Escherichia coli envelope.

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    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Vita N,Borne R,Fierobe HP

    更新日期:2020-03-01 00:00:00

  • Insect larval expression process is optimized by generating fusions with green fluorescent protein.

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    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Cha HJ,Dalal NG,Pham MQ,Vakharia VN,Rao G,Bentley WE

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

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    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


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

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    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Singhvi R,Stephanopoulos G,Wang DI

    更新日期:1994-04-05 00:00:00

  • Temperature-dependent dynamic control of the TCA cycle increases volumetric productivity of itaconic acid production by Escherichia coli.

    abstract::Based on the recently constructed Escherichia coli itaconic acid production strain ita23, we aimed to improve the productivity by applying a two-stage process strategy with decoupled production of biomass and itaconic acid. We constructed a strain ita32 (MG1655 ΔaceA Δpta ΔpykF ΔpykA pCadCs), which, in contrast to ita...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Harder BJ,Bettenbrock K,Klamt S

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  • Trehalose delays the reversible but not the irreversible thermal denaturation of cutinase.

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    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Baptista RP,Cabral JM,Melo EP

    更新日期:2000-12-20 00:00:00

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    pub_type: 杂志文章


    authors: Kang K,Ryu D,Drohan WN,Orthner CL

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    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Hathorne AP,Bermudez H

    更新日期:2013-07-01 00:00:00

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    abstract::During continuous lipase-catalyzed oleic acid esterification by ethanol in n-hexane, the oleic acid conversion, initially at 95%, decreases to 20% after 2 h. This decrease is caused by the accumulation of the water produced in the course of the reaction in the packed-bed reactor (PBR). In order to improve the PBR effi...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Colombié S,Tweddell RJ,Condoret JS,Marty A

    更新日期:1998-11-05 00:00:00

  • New tools for mass isotopomer data evaluation in (13)C flux analysis: mass isotope correction, data consistency checking, and precursor relationships.

    abstract::13C metabolic flux analysis (MFA) is based on carbon-labeling experiments where a specifically (13)C labeled substrate is fed. The labeled carbon atoms distribute over the metabolic network and the label enrichment of certain metabolic pools is measured by using different methods. Recently, MS methods have been dramat...

    journal_title:Biotechnology and bioengineering

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    authors: Wahl SA,Dauner M,Wiechert W

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    abstract::In addition to the role of maintaining the pH, buffers can also facilitate the transport of H(+) ions in acid-generating systems. The role of this facilitation in proton transport in porous pellets on acid-generating immobilized enzymic reactions is examined. The activity in these systems can be maximized by a proper ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Ruckenstein E,Rajora P

    更新日期:1985-06-01 00:00:00

  • Oil and eicosapentaenoic acid production by the diatom Phaeodactylum tricornutum cultivated outdoors in Green Wall Panel (GWP®) reactors.

    abstract::Phaeodactylum tricornutum is a widely studied diatom and has been proposed as a source of oil and polyunsaturated fatty acids (PUFA), particularly eicosapentaenoic acid (EPA). Recent studies indicate that lipid accumulation occurs under nutritional stress. Aim of this research was to determine how changes in nitrogen ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Rodolfi L,Biondi N,Guccione A,Bassi N,D'Ottavio M,Arganaraz G,Tredici MR

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    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Regmi P,Holgate B,Miller MW,Park H,Chandran K,Wett B,Murthy S,Bott CB

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  • Reactive oxygen species induced by shear stress mediate cell death in Bacillus subtilis.

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    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Sahoo S,Rao KK,Suraishkumar GK

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  • A study of D-lactate and extracellular methylglyoxal production in lactate re-utilizing CHO cultures.

    abstract::In large-scale mammalian cell culture, the key toxic by-products assessed and monitored are lactate and ammonia. Often no distinction between the two isoforms of lactate is made. Here, we present profiles of both D- and L-lactate. D-Lactate is the end molecule of the methylglyoxal pathway. D-Lactate unlike L-lactate i...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Paoli T,Faulkner J,O'kennedy R,Keshavarz-Moore E

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    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Mehrian M,Guyot Y,Papantoniou I,Olofsson S,Sonnaert M,Misener R,Geris L

    更新日期:2018-03-01 00:00:00

  • Kinetic analysis of deviation from the undifferentiated state in colonies of human induced pluripotent stem cells on feeder layers.

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    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Kim MH,Masuda E,Kino-oka M

    更新日期:2014-06-01 00:00:00

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    abstract::The effect of protein concentration in partitioning in PEG/salt aqueous two-phase systems has been investigated. PEG 4000/phosphate systems in the presence of 0% w/w and 8.8% w/w NaCl have been evaluated using amyloglucosidase, subtilisin, and trypsin inhibitor. Also, a PEG 4000/phosphate system with 3% w/w NaCl was u...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Schmidt AS,Andrews BA,Asenjo JA

    更新日期:1996-06-20 00:00:00

  • Use of stable emulsion to improve stability, activity, and enantioselectivity of lipase immobilized in a membrane reactor.

    abstract::The enantiocatalytic performance of immobilized lipase in an emulsion membrane reactor using stable emulsion prepared by membrane emulsification technology was studied. The production of optical pure (S)-naproxen from racemic naproxen methyl ester was used as a model reaction system. The O/W emulsion, containing the s...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Giorno L,Li N,Drioli E

    更新日期:2003-12-20 00:00:00

  • Gelatin interpenetration in poly N-isopropylacrylamide network reduces the compressive modulus of the scaffold: A property employed to mimic hepatic matrix stiffness.

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    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Sarkar J,Kamble SC,Patil R,Kumar A,Gosavi SW

    更新日期:2020-02-01 00:00:00

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    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Jungbauer A,Kaltenbrunner O

    更新日期:1996-10-20 00:00:00

  • Enhanced d-lactic acid production by recombinant Saccharomyces cerevisiae following optimization of the global metabolic pathway.

    abstract::Utilization of renewable feedstocks for the production of bio-based chemicals such as d-lactic acid by engineering metabolic pathways in the yeast Saccharomyces cerevisiae has recently become an attractive option. In this study, to realize efficient d-lactic acid production by S. cerevisiae, the expression of 12 glyco...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Yamada R,Wakita K,Mitsui R,Ogino H

    更新日期:2017-09-01 00:00:00