Green fluorescent protein in Saccharomyces cerevisiae: real-time studies of the GAL1 promoter.


:Green fluorescent protein (GFP) was used to study the regulation of the galactose-inducible GAL1 promoter in yeast Saccharomyces cerevisiae strains. GFP was cloned into the pGAL110 vector and transformed into the yeast strains. Time course studies comparing culture fluorescence intensity and GFP concentration were conducted along with on-line monitoring of GFP expression. Our results demonstrated that GFP fluorescence could be used as a quantifiable on-line reporter gene in yeast strains. The effect of an integrated GAL10p-GAL4 transcription cassette was investigated. Induction time studies showed that there was no significant difference in GFP expression level by adding galactose at different culture times. A wide range of galactose concentrations was used to study the initial galactose concentration effect on GFP expression kinetics. A minimum of 0.05 g/L galactose doubled the GFP fluorescence signal as compared to the control, whereas 0.1 g/L gave the highest specific GFP yield. A simple analytical model was proposed to describe GFP expression kinetics based on the experimental results. In addition, this GFP-based approach was shown to have potential use for high-throughput studies. The use of GFP as a generic tool provided important insights to the GAL expression system and has great potential for further process optimization applications.


Biotechnol Bioeng


Li J,Wang S,VanDusen WJ,Schultz LD,George HA,Herber WK,Chae HJ,Bentley WE,Rao G




Has Abstract


2000-10-20 00:00:00














  • Steam pretreatment of lignocellulosic material for enhanced enzymatic hydrolysis.

    abstract::Pretreatment methods were compared with steam explosion, and differing views on the relative importance of mechanical and chemical effects were outlined. Hydrolysis was desirable; pyrolysis was undesirable. The effects of initial moisture content on steam consumption, mechanism and rate of heat transfer, pentosan solu...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Brownell HH,Saddler JN

    更新日期:1987-02-01 00:00:00

  • Continuous ethanol production and cell growth in an immobilized-cell bioreactor employing Zymomonas mobilis.

    abstract::Ethanol-producing bioreactors employing cells of Zymomonas mobilis attached to glass-fiber pads were operated continuously for as long as 28 days. Ethanol production, which is related to bed-associated biomass levels, was found to occur in three distinct phases: an exponential phase, a linear phase, and a "steady-stat...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Arcuri EJ

    更新日期:1982-03-01 00:00:00

  • Novel, linked bioreactor system for continuous production of biologics.

    abstract::A novel, alternative intensified cell culture process comprised of a linked bioreactor system is presented. An N-1 perfusion bioreactor maintained cells in a highly proliferative state and provided a continuous inoculum source to a second bioreactor operating as a continuous-flow stirred-tank reactor (CSTR). An initia...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Gagnon M,Nagre S,Wang W,Coffman J,Hiller GW

    更新日期:2019-08-01 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

    更新日期:2004-05-05 00:00:00

  • On the pH memory of lyophilized compounds containing protein functional groups.

    abstract::Lyophilized proteins exhibit "pH memory," i.e., their behavior in the solid form corresponds to the pH of the aqueous solution from which they were freeze dried. Herein, we directly tested whether the ionization state is "remembered" by model organic compounds containing various protein functional groups (amino, carbo...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Costantino HR,Griebenow K,Langer R,Klibanov AM

    更新日期:1997-02-05 00:00:00

  • Growth of Trichosporon cutaneum under oxygen limitation: kinetics of oxygen uptake.

    abstract::The relationship between oxygen concentration and growth rate in the yeast Trichosporon cutaneum was studied. In order to establish the conditions for purely oxygen-limited growth, the cells were first grown in a carbon-limited chemostat, and kinetic parameters determined. The cells were then grown in an oxygen-limite...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Käppeli O,Fiechter A

    更新日期:1982-11-01 00:00:00

  • Fermentation and isolation of C10-deacetylase for the production of 10-deacetylbaccatin III from baccatin III.

    abstract::10-Deacetylabaccatin III (10 DAB), an important precursor for paclitaxel semisynthesis, is enhanced in yew extracts using C10-deacetylase and C13-deacylase enzymes.(4) C10-deacetylase is an intracellular enzyme produced by the fermentation of a soil microorganism, Nocardioides luteus (SC 13912). During the fermentatio...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Nanduri VB,Hanson RL,Laporte TL,Ko RY,Patel RN,Szarka LJ

    更新日期:1995-12-05 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

    更新日期:2017-10-01 00:00:00

  • Regulation of lac operon expression in mixed sugar chemostat cultures.

    abstract::Expression of the lactose (lac) operon in the Escherichia coli chromosome has been studied in mixed-sugar chemostat cultures under steady-state and transient conditions. A unified model has been formulated which involves regulation of active inducer (lactose) transport, promoter-operator regulated expression of the la...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Ray NG,Vieth WR,Venkatasubramanian K

    更新日期:1987-06-01 00:00:00

  • Countercurrent multistage fluidized bed reactor for immobilized biocatalysts: I. Modeling and simulation.

    abstract::For the application of immobilized enzymes, fixed bed reactors are used almost exclusively. Fixed bed reactors have specific disadvantages, especially for processes with a deactivating catalyst. Therefore, we have studied a novel reactor type with continuous transport of the immobilized biocatalyst. Flow of biocatalys...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Vos HJ,Groen DJ,Potters JJ,Luyben KC

    更新日期:1990-08-05 00:00:00

  • Characterization of hybridoma cell responses to elevated pCO(2) and osmolality: intracellular pH, cell size, apoptosis, and metabolism.

    abstract::CO(2) partial pressure (pCO(2)) in industrial cell culture reactors may reach 150-200 mm Hg, which can significantly inhibit cell growth and recombinant protein production. The inhibitory effects of elevated pCO(2) at constant pH are due to a combination of the increases in pCO(2) and [HCO(-) (3)], per se, and the ass...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: deZengotita VM,Schmelzer AE,Miller WM

    更新日期:2002-02-15 00:00:00

  • Unified kinetic treatment for growth on dual nutrients.

    abstract::This paper presents a new formulation for the analysis of growth kinetics on multiple nutrients. The baseline for the theory is the concept that a noninteractive growth process occurs among perfectly substitutable nutrients if the locus of points of the substrate concentrations producing equal growth rate is linear. A...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Haas CN

    更新日期:1994-06-20 00:00:00

  • Metabolic engineering of Ralstonia eutropha for the production of polyhydroxyalkanoates from sucrose.

    abstract::A sucrose utilization pathway was established in Ralstonia eutropha NCIMB11599 and R. eutropha 437-540 by introducing the Mannheimia succiniciproducens MBEL55E sacC gene that encodes β-fructofuranosidase. These engineered strains were examined for the production of poly(3-hydroxybutyrate) [P(3HB)] and poly(3-hydroxybu...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Park SJ,Jang YA,Noh W,Oh YH,Lee H,David Y,Baylon MG,Shin J,Yang JE,Choi SY,Lee SH,Lee SY

    更新日期:2015-03-01 00:00:00

  • Preparation and properties of soluble-insoluble immobilized proteases.

    abstract::In order to carry out an effective enzyme reaction, the preparation of soluble-insoluble immobilized enzyme was investigated. Proteases were selected as model enzymes, and their immobilization was carried out by using an enteric coating polymer as a carrier. Among the polymers tested, methacrylic acid-methylacrylate-m...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Fujimura M,Mori T,Tosa T

    更新日期:1987-04-01 00:00:00

  • Analysis of E. coli phoA-lacZ fusion gene expression inserted into a multicopy plasmid and host cell's chromosome.

    abstract::The production of beta-galactosidase from the E. coli phoA-lacZ fusion gene was studied to compare the gene expression behavior of two cloning methods: insertion to multicopy plasmids and integration into host cell's chromosome. The chromosome-integrating strain showed more tight control of fusion gene expression leve...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Shin PK,Seo JH

    更新日期:1990-12-20 00:00:00

  • Microcalorimetric study of cellulose degradation by Cellulomonas uda ATCC 21399.

    abstract::A newly designed batch calorimeter was used to investigate the degradability of some celluloses having varying degrees of crystallinity. The PTC of an aerobic culture of Cellulomonas uda ATCC 21399 obtained revealed a diauxic growth which is attributed to the presence of hemicellulose contaminating Avicel and MN300 ce...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Dermoun Z,Belaich JP

    更新日期:1985-07-01 00:00:00

  • Simultaneous detection of three sex steroid hormone classes using a novel yeast-based biosensor.

    abstract::A biosensor detecting estrogens, progestogens, and androgens in complex samples and in a single step is described. Three Arxula adeninivorans yeast strains were created, each strain producing a different recombinant human hormone receptor and a different fluorescent reporter protein. These strains were then mixed to c...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Chamas A,Pham HTM,Jähne M,Hettwer K,Uhlig S,Simon K,Einspanier A,Baronian K,Kunze G

    更新日期:2017-07-01 00:00:00

  • Mass transport parameters of aspen wood chip beds via stimulus-response tracer techniques.

    abstract::A stimulus-response tracer technique has been used to characterize packed beds of untreated, as well as acid prehydrolyzed, and enzymatically hydrolyzed aspen wood chips. Glucose was used as the tracer. Bulk liquid phase dispersion, interphase mass transfer, and intraparticle diffusion coefficients were determined for...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Hradil G,Calo JM,Wunderlich TK Jr

    更新日期:1993-02-05 00:00:00

  • Escherichia coli "TatExpress" strains super-secrete human growth hormone into the bacterial periplasm by the Tat pathway.

    abstract::Numerous high-value proteins are secreted into the Escherichia coli periplasm by the General Secretory (Sec) pathway, but Sec-based production chassis cannot handle many potential target proteins. The Tat pathway offers a promising alternative because it transports fully folded proteins; however, yields have been too ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Browning DF,Richards KL,Peswani AR,Roobol J,Busby SJW,Robinson C

    更新日期:2017-12-01 00:00:00

  • Non-invasive online detection of microbial lysine formation in stirred tank bioreactors by using calorespirometry.

    abstract::Non-invasive methods for online monitoring of biotechnological processes without compromising the integrity of the reactor system are very important to generate continuous data. Even though calorimetry has been used in conventional biochemical analysis for decades, it has not yet been specifically applied for online d...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Regestein L,Maskow T,Tack A,Knabben I,Wunderlich M,Lerchner J,Büchs J

    更新日期:2013-05-01 00:00:00

  • Purification of recombinant protein by cold-coacervation of fusion constructs incorporating resilin-inspired polypeptides.

    abstract::Polypeptides containing between 4 and 32 repeats of a resilin-inspired sequence AQTPSSYGAP, derived from the mosquito Anopheles gambiae, have been used as tags on recombinant fusion proteins. These repeating polypeptides were inspired by the repeating structures that are found in resilins and sequence-related proteins...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Lyons RE,Elvin CM,Taylor K,Lekieffre N,Ramshaw JA

    更新日期:2012-12-01 00:00:00

  • Epi-CHO, an episomal expression system for recombinant protein production in CHO cells.

    abstract::This study describes the development of a transient expression system for CHO cells based on autonomous replication and retention of transfected plasmid DNA. A transient expression system that allows extrachromosomal amplification of plasmids permits more plasmid copies to persist in the transfected cell throughout th...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Kunaparaju R,Liao M,Sunstrom NA

    更新日期:2005-09-20 00:00:00

  • Enzymatic glucosylation of hydrophobic alcohols in organic medium by the reverse hydrolysis reaction using almond-beta-D-glucosidase.

    abstract::Alkyl beta-D-glucosides were synthesized from D-glucose and alcohols by reverse hydrolysis using the commercially available almond beta-D-glucosidase in 9:1 (v/v) acetonitrile-water medium. The main characteristics of this enzyme-catalyzed glucosylation were established by using 2-hydroxybenzyl alcohol. The reaction i...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Vic G,Biton J,Le Beller D,Michel JM,Thomas D

    更新日期:1995-04-20 00:00:00

  • Enhancement of ethanol and biogas production from high-crystalline cellulose by different modes of NMO pretreatment.

    abstract::Pretreatment of high-crystalline cellulose with N-methyl-morpholine-N-oxide (NMO or NMMO) to improve bioethanol and biogas production was investigated. The pretreatments were performed at 90 and 120 degrees C for 0.5-15 h in three different modes, including dissolution (85% NMO), ballooning (79% NMO), and swelling (73...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Jeihanipour A,Karimi K,Taherzadeh MJ

    更新日期:2010-02-15 00:00:00

  • Combining metabolic engineering and metabolic evolution to develop nonrecombinant strains of Escherichia coli C that produce succinate and malate.

    abstract::Derivatives of Escherichia coli C were engineered to produce primarily succinate or malate in mineral salts media using simple fermentations (anaerobic stirred batch with pH control) without the addition of plasmids or foreign genes. This was done by a combination of gene deletions (genetic engineering) and metabolic ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Jantama K,Haupt MJ,Svoronos SA,Zhang X,Moore JC,Shanmugam KT,Ingram LO

    更新日期:2008-04-01 00:00:00

  • Selective adsorption of plant products.

    abstract::The results from this study demonstrate that neutral resins can selectively concentrate specific indole alkaloids from dilute aqueous mixtures. Adsorption was observed to provide a one to two order of magnitude improvement in concentrating these alkaloids, as compared to an equivalent single-staged extraction. Since t...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Payne GF,Shuler ML

    更新日期:1988-06-05 00:00:00

  • Identification and control of novel growth inhibitors in fed-batch cultures of Chinese hamster ovary cells.

    abstract::Chinese hamster ovary (CHO) cells in culture are known to consume large amounts of nutrients and divert most of them toward byproducts, some of which, including lactate and ammonia, are known to be toxic in nature. Glucose limitation strategies can successfully control lactate accumulation in fed-batch cultures yieldi...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Mulukutla BC,Kale J,Kalomeris T,Jacobs M,Hiller GW

    更新日期:2017-08-01 00:00:00

  • Simultaneous saccharification and co-fermentation of paper sludge to ethanol by Saccharomyces cerevisiae RWB222. Part II: investigation of discrepancies between predicted and observed performance at high solids concentration.

    abstract::The simultaneous saccharification and co-fermentation (SSCF) kinetic model described in the companion paper can predict batch and fed batch fermentations well at solids concentrations up to 62.4 g/L cellulose paper sludge but not in batch fermentation at 82.0 g/L cellulose paper sludge. Four hypotheses for the discrep...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Zhang J,Shao X,Lynd LR

    更新日期:2009-12-01 00:00:00

  • Design of reactive porous polymer supports for high throughput bioreactors: poly(2-vinyl-4,4-dimethylazlactone-co-acrylamide- co-ethylene dimethacrylate) monoliths.

    abstract::Enzymatic bioreactors with both high flow characteristics and mechanical stability based on macroporous poly(2-vinyl-4, 4-dimethylazlactone-co-acrylamide-co-ethylene dimethacrylate) monoliths have been prepared. Covalent immobilization of trypsin on these support is achieved in a single reaction step using the azlacto...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Xie S,Svec F,Fréchet JM

    更新日期:1999-01-05 00:00:00

  • Cellulase production by trichoderma harzianum in static and mixed solid-state fermentation reactors under nonaseptic conditions.

    abstract::Cellulase production from lignocellulosic materials was studied in solid-state cultivation by both static and mixed techniques under nonaseptic conditions. The effects of fermentation conditions, such as moisture content, pH, temperature, and aeration, on cellulase production by Trichoderma harzianum using a mixture o...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Deschamps F,Giuliano C,Asther M,Huet MC,Roussos S

    更新日期:1985-09-01 00:00:00