Optimization of amorphadiene synthesis in bacillus subtilis via transcriptional, translational, and media modulation.


:Genetically engineered microbes have been intensively investigated as a means for the cost-effective production of isoprenoids. Bacillus subtilis is a promising microbial host for this purpose because of its fast growth rate and GRAS (generally regarded as safe) status. To date, development of this host has been impaired by the lack of genetic tools for modulating the expression of multiple genes. In this study, we present a novel two-promoter system which can be used to independently control the expression levels of two gene cassettes over a large dynamic range. Coupled with protein translation engineering and systematic media optimization, ~20 mg/L amorphadiene was produced in shake flask scale, a 40-fold improvement over the highest reported isoprenoid product yield in B. subtilis. As the tools and strategies developed here can be extended to the overproduction of other valuable metabolites, this proof-of-concept study lays the foundation for high level heterologous production of isoprenoids in Bacillus.


Biotechnol Bioeng


Zhou K,Zou R,Zhang C,Stephanopoulos G,Too HP




Has Abstract


2013-09-01 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


    authors: Hosseinzadegan H,Tafti DK

    更新日期:2017-10-01 00:00:00

  • The use of the maharanobis and modified distances for the improvement of simulation of glutamic acid production.

    abstract::A modified simulation procedure based on a statistical approach was investigated. The procedure predicts the time course of fed-batch culture for glutamic acid production by a temperature-sensitive strain of Brevibacterium flavum. The statistical approach requires only a data base of state points obtained in experimen...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Kishimoto M,Alfafara CG,Nakajima M,Yoshida T,Taguchi H

    更新日期:1989-01-10 00:00:00

  • Expanded bed adsorption/desorption of proteins with Streamline Direct CST I adsorbent.

    abstract::Streamline Direct CST I is a new type of ion exchanger with multi-modal functional groups, specially designed for an expanded bed adsorption (EBA) process, which can capture directly the proteins from the high ionic strength feedstocks with a high binding capacity. In this study, an experimental study is carried out f...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Li P,Xiu G,Mata VG,Grande CA,Rodrigues AE

    更新日期:2006-08-20 00:00:00

  • Quantitative image analysis as a diagnostic tool for monitoring structural changes of anaerobic granular sludge during detergent shock loads.

    abstract::Two shock loads of a commercial detergent (I-150 mg chemical oxygen demand (COD)/L, fed for 56 h; II-300 mg COD/L fed for 222 h) were applied in a lab-scale Expanded Granular Sludge Blanket (EGSB) reactor, fed with 1,500 mg COD/L of ethanol. The impact of the surfactant was assessed in terms of granular sludge morphol...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Costa JC,Abreu AA,Ferreira EC,Alves MM

    更新日期:2007-09-01 00:00:00

  • Permeability of a growing biofilm in a porous media fluid flow analyzed by magnetic resonance displacement-relaxation correlations.

    abstract::Biofilm growth in porous media is difficult to study non-invasively due to the opaqueness and heterogeneity of the systems. Magnetic resonance is utilized to non-invasively study water dynamics within porous media. Displacement-relaxation correlation experiments were performed on fluid flow during biofilm growth in a ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Vogt SJ,Sanderlin AB,Seymour JD,Codd SL

    更新日期:2013-05-01 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

  • Immobilization of thermolysin to polyamide nonwoven materials.

    abstract::In the last few years, an increasing number of biotechnological techniques have been applied to the restoration and conservation of works of art, paintings, old maps, and papers or books. Enzymes can solve problems that give restorers difficulties, although for many applications it is not possible to use soluble enzym...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Moeschel K,Nouaimi M,Steinbrenner C,Bisswanger H

    更新日期:2003-04-20 00:00:00

  • Harvesting of Scenedesmus obliquus in wastewaters: auto- or bioflocculation?

    abstract::Autoflocculation and bioflocculation are considered to be the most promising means for the economical harvesting of microalgae. We have therefore studied these phenomena with cultures of Scenedesmus obliquus produced during biological tertiary wastewater treatment. The quantity of extracellular polymers produced durin...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Lavoie A,de la Noüe J

    更新日期:1987-11-01 00:00:00

  • Effect of gene copy number and chaperone coexpression on recombinant hydrophobin HFBI biosurfactant production in Pichia pastoris.

    abstract::Hydrophobins are small highly surface-active fungal proteins with potential as biosurfactants in a wide array of applications. However, practical implementation of hydrophobins at large scale has been hindered by low recombinant yields. In this study, the effects of increasing hydrophobin gene copy number and overexpr...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Sallada ND,Harkins LE,Berger BW

    更新日期:2019-08-01 00:00:00

  • The immobilizaton of enzymes, bovine serum albumin, and phenylpropylamine to poly(acrylic acid)-polyethylene-based copolymers.

    abstract::A poly(acrylic acid)-polyethylene graft copolymer was prepared and used initially to couple to acid phosphatase, using soluble carbodiimides. Yields which were quite good were obtained with CMC but not with EDAC. The copolymers was used to couple trypsin using EEDQ. Several organic solvents were investigated for the p...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Beddows CG,Gil MH,Guthrie JT

    更新日期:1982-06-01 00:00:00

  • Effect of non-feeding period length on the intermittent operation of UASB reactors treating dairy effluents.

    abstract::Recent environmental concerns have prompted a re-evaluation of conventional management strategies and refueled the search of innovative waste management practices. In this sense, the anaerobic digestion of both fat and the remaining complex organic matter present in dairy wastewaters is attractive, although the contin...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Coelho NM,Rodrigues AA,Arroja LM,Capela IF

    更新日期:2007-02-01 00:00:00

  • In situ determination of kinetic parameters for biofilms: Isolation and characterization of oligotrophic biofilms.

    abstract::This article develops and utilizes an in situ technique to estimate the Monod half-maximum rate concentration, K(s) and the maximum specific utilization rate constant, k, for biofilms. The technique employs a curve-matching method with kinetic results from several short-term experiments with completely mixed biofilm r...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Rittmann BE,Crawford L,Tuck CK,Namkung E

    更新日期:1986-11-01 00:00:00

  • Diffusion coefficients of glucose and ethanol in cell-free and cell-occupied calcium alginate membranes.

    abstract::The diffusivities of glucose and ethanol in cell-free and cell-occupied membranes of calcium alginate were measured in a diffusion cell. The lag time analysis was used. Diffusivities decreased with increasing alginate concentration and were comparable with those in water for a 2% alginate membrane. Glucose and ethanol...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Hannoun BJ,Stephanopoulos G

    更新日期:1986-06-01 00:00:00

  • Optimization of rifamycin B fermentation in shake flasks via a machine-learning-based approach.

    abstract::Rifamycin B is an important polyketide antibiotic used in the treatment of tuberculosis and leprosy. We present results on medium optimization for Rifamycin B production via a barbital insensitive mutant strain of Amycolatopsis mediterranei S699. Machine-learning approaches such as Genetic algorithm (GA), Neighborhood...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Bapat PM,Wangikar PP

    更新日期:2004-04-20 00:00:00

  • Dibutyl phthalate biodegradation by the white rot fungus, Polyporus brumalis.

    abstract::In this study, white rot fungus, Polyporus brumalis, was applied to degrade dibutyl phthalate (DBP), a major environmental pollutant. The degradation potential and resulting products were evaluated with HPLC and GC/MS. As DBP concentration increased to 250, 750, and 1,250 microM, the mycelial growth of P. brumalis was...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Lee SM,Lee JW,Koo BW,Kim MK,Choi DH,Choi IG

    更新日期:2007-08-15 00:00:00

  • Growth and fermentation characteristics of new selected strains of Saccharomyces at high temperatures and high cell densities.

    abstract::Maintenance of high cell viability was the main characteristic of our new strains of thermotolerant Saccharomyces. Total sugar conversion to ethanol was observed for sugarcane juice fermentation at 38-40 degrees C in less than 10 h and without continuous aeration of the culture. Invertase activity differed among the s...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Laluce C,Palmieri MC,Lopes da Cruz RC

    更新日期:1991-03-15 00:00:00

  • Diffusion of lactate and ammonium in relation to growth of Geotrichum candidum at the surface of solid media.

    abstract::Geotrichum candidum was cultivated at the surface of solid model media containing peptone to simulate the composition of Camembert cheese. The surface growth of G. candidum induced the diffusion of substrates from the core to the rind and the diffusion of produced metabolites from the rind to the core. In the range of...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Aldarf M,Fourcade F,Amrane A,Prigent Y

    更新日期:2004-07-05 00:00:00

  • Light-responsive bioconjugates as novel tools for specific capture of biologicals by photoaffinity precipitation.

    abstract::Light-responsive bioconjugates are synthesized by a two-step protocol calling first for cotelomerization (chain-transfer polymerization) of N-isopropylacrylamide and N-acryloxysuccinimide. The desired bioligand (biotin) is used in modified form as chain-transfer agent in this step. As a consequence, 100% of the produc...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Desponds A,Freitag R

    更新日期:2005-09-05 00:00:00

  • Membrane-attached biofilms for VOC wastewater treatment. II: Effect of biofilm thickness on performance.

    abstract::This article reports a study of the performance of membrane-attached biofilms grown in a single tube extractive membrane bioreactor (STEMS) used for the treatment of a synthetic wastewater containing a toxic VOC (1,2-dichloroethane [DCE]). Mass balances show that complete mineralization of DCE was achieved, and that t...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Dos Santos LM,Livingston AG

    更新日期:1995-07-05 00:00:00

  • Kinetics and mass-transfer phenomena in anaerobic granular sludge.

    abstract::The kinetic properties of acetate-degrading methanogenic granular sludge of different mean diameters were assessed at different up-flow velocities (V(up)). Using this approach, the influence of internal and external mass transfer could be estimated. First, the apparent Monod constant (K(S)) for each data set was calcu...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Gonzalez-Gil G,Seghezzo L,Lettinga G,Kleerebezem R

    更新日期:2001-04-20 00:00:00

  • Biofilm thickness variability investigated with a laser triangulation sensor.

    abstract::Measurement of the surface roughness and thickness of biological films is laborious and usually destructive, thus hampering research in this area. We developed a laser triangulation sensor (LTS) set-up for the fast and nondestructive measurement of these biofilm parameters during growth. Using LTS measurements, the mo...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Okkerse WJ,Ottengraf SP,Osinga-Kuipers B

    更新日期:2000-12-20 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

  • A novel convenient procedure for extractive work-up of whole-cell biotransformations using de-emulsifying hydrolases.

    abstract::Extractive work-up of whole-cell biotransformations generally suffers from the formation of stable gels and slimes upon addition of the organic solvent to the cell suspension and the cell-free solution, respectively. This problem has been overcome by enzymatic lysis of emulsifying agents present in the medium through ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Jörg G,Leppchen K,Daussmann T,Bertau M

    更新日期:2004-08-20 00:00:00

  • Effects of calcium on development of anaerobic acidogenic biofilms.

    abstract::Anerobic biofilms with dominantly acidogenic bacteria were grown in fixed-bed recycle reactors. The influence of calcium concentration in the culture medium on biofilm mass accumulation, immobilized calcium concentration, and biofilm-specific activity was investigated. The results indicate that the biofilm mass accumu...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Huang J,Pinder KL

    更新日期:1995-02-05 00:00:00

  • Individual and combined effects of the rolA, B, and C genes on anthraquinone production in Rubia cordifolia transformed calli.

    abstract::It is known that the rolA, rolB, and rolC genes of Agrobacterium rhizogenes T-DNA affect processes of plant development and activate the synthesis of secondary metabolites in transformed plant cells. Although a synergistic activity of the rol genes on root formation is well-documented, little is known about their indi...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Shkryl YN,Veremeichik GN,Bulgakov VP,Tchernoded GK,Mischenko NP,Fedoreyev SA,Zhuravlev YN

    更新日期:2008-05-01 00:00:00

  • Oxygen uptake rates and liver-specific functions of hepatocyte and 3T3 fibroblast co-cultures.

    abstract::Bioartificial liver (BAL) devices have been developed to treat patients undergoing acute liver failure. One of the most important parameters to consider in designing these devices is the oxygen consumption rate of the seeded hepatocytes which are known to have oxygen consumption rates 10 times higher than most other c...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Cho CH,Park J,Nagrath D,Tilles AW,Berthiaume F,Toner M,Yarmush ML

    更新日期:2007-05-01 00:00:00

  • An exclusion mechanism in ion exchange chromatography.

    abstract::Protein dynamic binding capacities on ion exchange resins are typically expected to decrease with increasing conductivity and decreasing protein charge. There are, however, conditions where capacity increases with increasing conductivity and decreasing protein charge. Capacity measurements on two different commercial ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Harinarayan C,Mueller J,Ljunglöf A,Fahrner R,Van Alstine J,van Reis R

    更新日期:2006-12-05 00:00:00

  • Metabolic engineering of acid resistance elements to improve acid resistance and propionic acid production of Propionibacterium jensenii.

    abstract::Propionic acid (PA) and its salts are widely used in the food, pharmaceutical, and chemical industries. Microbial production of PA by propionibacteria is a typical product-inhibited process, and acid resistance is crucial in the improvement of PA titers and productivity. We previously identified two key acid resistanc...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Guan N,Li J,Shin HD,Du G,Chen J,Liu L

    更新日期:2016-06-01 00:00:00

  • Behavior of spores of Penicillium roquefortii during fed-batch bioconversion of octanoic acid into 2-heptanone.

    abstract::The bioconversion of octanoic acid into 2-heptanone by spores of Penicillium roquefortii is performed using a fed-batch technique with pH control by addition of the liquid substrate itself. The early stage of this process takes place with a high bioconversion rate and high yield. These values then decrease as a result...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Larroche C,Besson I,Gros JB

    更新日期:1994-09-05 00:00:00

  • Simulation of penicillin production in fed-batch cultivations using a morphologically structured model.

    abstract::A mathematical model is formulated to describe trends in biomass and penicillin formation as well as substrate consumption for fed-batch cultivations. The biomass is structured into three morphological compartments, and glucose and corn steep liquor are considered as substrates for growth. Penicillin formation is assu...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章


    authors: Zangirolami TC,Johansen CL,Nielsen J,Jørgensen SB

    更新日期:1997-12-20 00:00:00