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

Abstract:

: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.

journal_name

Biotechnol Bioeng

authors

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

doi

10.1002/bit.24900

subject

Has Abstract

pub_date

2013-09-01 00:00:00

pages

2556-61

issue

9

eissn

0006-3592

issn

1097-0290

journal_volume

110

pub_type

杂志文章
  • Comparative metabolic profiling revealed limitations in xylose-fermenting yeast during co-fermentation of glucose and xylose in the presence of inhibitors.

    abstract::During lignocellulosic ethanol fermentation, yeasts are exposed to various lignocellulose-derived inhibitors, which disrupt the efficiency of hexose and pentose co-fermentation. To understand the metabolic response of fermentation microbes to these inhibitors, a comparative metabolomic investigation was performed on a...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.24992

    authors: Wang X,Jin M,Balan V,Jones AD,Li X,Li BZ,Dale BE,Yuan YJ

    更新日期:2014-01-01 00:00:00

  • A continuous membrane bioreactor for ester synthesis in organic media: I. Operational characterization and stability.

    abstract::The feasibility of continuous ester synthesis in a membrane bioreactor (MBR) by a recombinant cutinase from Fusarium solani pisi was investigated, using the optimal conditions previously attained by medium engineering. The objective was to analyze the MBR behavior as a differential or an integral reactor. The main com...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/1097-0290(20010120)72:2<127::aid-bit1>3.0.

    authors: Carvalho CM,Aires-Barros MR,Cabral JM

    更新日期:2001-01-20 00:00:00

  • Rapid construction and characterization of synthetic antibody libraries without DNA amplification.

    abstract::We report on a simple method to rapidly generate very large libraries of genes encoding mutant proteins without the use of DNA amplification, and the application of this methodology in the construction of synthetic immunoglobulin variable heavy (V(H)) and light (V(kappa)) libraries. Four high quality, chemically synth...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.22712

    authors: Ge X,Mazor Y,Hunicke-Smith SP,Ellington AD,Georgiou G

    更新日期:2010-06-15 00:00:00

  • A kinetic model for methanogenesis from whey permeate in a packed bed immobilized cell bioreactor.

    abstract::The fermentation kinetics of methane production from whey permeate in a packed bed immobilized cell bioreactor at mesophilic temperatures and pHs around neutral was studied. Propionate and acetate were the only two major organic intermediates found in the methanogenic fermentation of lactose. Based on this finding, a ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260370412

    authors: Yang ST,Quo M

    更新日期:1991-02-20 00:00:00

  • Yeast suspension filtration: flux enhancement using an upward gas/liquid slug flow-application to continuous alcoholic fermentation with cell recycle.

    abstract::This study deals with the use of an upward gas/liquid slug flow to reduce tubular mineral membrane fouling. The injection of air into the feedstream is designed to create hydrodynamic conditions that destabilize the cake layer over the membrane surface inside the filtration module complex. Experimental study was carri...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/(sici)1097-0290(19980405)58:1<47::aid-bit5

    authors: Mercier M,Maranges C,Fonade C,Lafforgue-Delorme C

    更新日期:1998-04-05 00:00:00

  • Measurement of trans-epithelial electrical resistance in perfusion: potential application for in vitro ocular toxicity testing.

    abstract::An in vitro toxicity testing system that combines perfusion of a membrane-supported tissue model with measurement of trans-epithelial electrical resistance (TER) is described. The system is fully automated and allows for continuous TER measurements during exposure to a test agent. The utility of this system is demonst...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/(SICI)1097-0290(19960605)50:5<568::AID-BIT

    authors: Pasternak AS,Miller WM

    更新日期:1996-06-05 00:00:00

  • Influence of uncertainties in pH, pMg, activity coefficients, metabolite concentrations, and other factors on the analysis of the thermodynamic feasibility of metabolic pathways.

    abstract::Thermodynamic feasibility analysis (TFA) has been used as a tool capable of providing additional constraints to the mass balance-based methods of analysis of metabolic networks (e.g., flux balance analysis). Several publications have recently appeared in which TFA of different metabolic pathways from relatively simple...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.22309

    authors: Vojinović V,von Stockar U

    更新日期:2009-07-01 00:00:00

  • Mass transfer parameters in an immobilized glucoamylase column by pulse response analysis.

    abstract::Pulses of several nonreacting solutes were fed to a column containing immobilized enzyme catalyst beads. The response curves were recorded, and their first and second moments were determined by numerical integration. From the moment analysis, the void volume, internal porosity, and tortuosity of the support were calcu...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260250707

    authors: Sirotti DA,Emery A

    更新日期:1983-07-01 00:00:00

  • Detachment of biomass from suspended nongrowing spherical biofilms in airlift reactors.

    abstract::In three-phase internal loop airlift reactors, the detachment of biomass from suspended biofilm pellets in the presence of bare carrier particles was investigated under nongrowth conditions. The detachment rate was dominated by collisions between bare carrier particles and biofilm pellets. The concentration of bare ca...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260460309

    authors: Gjaltema A,Tijhuis L,van Loosdrecht MC,Heijnen JJ

    更新日期:1995-05-05 00:00:00

  • Optimization of a reactor assembly for the production of 6-APA from penicillin-V.

    abstract::The design and operation of an industrial penicillin-V deacylation reactor is simulated, using a kinetic expression and mass transport parameters for the immobilized enzyme particles which were determined experimentally in a previous study. It is desirable to use a series of equalsized plug flow reactors with pH contr...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260261213

    authors: Karlsen LG,Villadsen J

    更新日期:1984-12-01 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

    doi:10.1002/bit.260270711

    authors: Dermoun Z,Belaich JP

    更新日期:1985-07-01 00:00:00

  • Bioartificial kidney. I. Theoretical analysis of convective flow in hollow fiber modules: application to a bioartificial hemofilter.

    abstract::Analytical expressions describing convective flow in a continuous arteriovenous hollow fiber hemofilter were developed. In the lumen of the hollow fiber membrane, existing analytical expressions were applied to describe velocity profiles and pressure. For flow in the shell (the extracapillary space separating the fibe...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:

    authors: Moussy Y

    更新日期:2000-04-20 00:00:00

  • Review: tissue engineering: reconstitution of human hematopoiesis ex vivo.

    abstract::The reconstruction of functioning human tissues ex vivo is becoming an important part of biotechnology. There are compelling scientific, clinical, and biotechnological reasons for fully or partially reconstituting human tissues such as skin, bone marrow, and liver ex vivo. In particular, bone marrow is a tissue of muc...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260420802

    authors: Koller MR,Palsson BO

    更新日期:1993-10-01 00:00:00

  • Enhanced purification of plasmid DNA isoforms by exploiting ionic strength effects during ultrafiltration.

    abstract::The solution structure of plasmid DNA is known to be a strong function of solution conditions due to intramolecular electrostatic interactions between the charged phosphate groups along the DNA backbone. The objective of this work was to determine whether it was possible to enhance the use of ultrafiltration for separ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.25836

    authors: Li Y,Currie D,Zydney AL

    更新日期:2016-04-01 00:00:00

  • Single-step unimolecular non-first-order enzyme deactivation kinetics.

    abstract::A two-parameter deactivation model is proposed to describe the kinetics of activity stabilization for some enzymes. The single-step unimolecular mechanism exhibits non-first-order deactivation kinetics since the final enzyme state, E(1) is not completely inactivated. The usefulness of the model is demonstrated by appl...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260300604

    authors: Sadana A,Henley JP

    更新日期:1987-10-20 00:00:00

  • Mitigation of ammonia inhibition by internal dilution in high-rate anaerobic digestion of food waste leachate and evidences of microbial community response.

    abstract::A high-rate anaerobic digestion of food waste leachate were tested using intermittent continuously stirred tank reactors (iCSTRs) to evaluate how severe ammonia inhibition could be mitigated with internal dilution strategy, and to identify how bacterial and archaeal community respond in genus and species level. Experi...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.25968

    authors: Yun YM,Kim DH,Cho SK,Shin HS,Jung KW,Kim HW

    更新日期:2016-09-01 00:00:00

  • Gas transfer characteristics of a novel membrane bioreactor.

    abstract::We show the design features of a membrane bioreactor based on pulsatile flow across dimpled membranes. Results show an enhanced mass transfer of air of at least five-fold magnitude as compared with flat membranes. An increased working volume form 20 mL to 120 mL reduced the k(L)A at a given Reynolds number because of ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260381016

    authors: Beeton S,Millward HR,Bellhouse BJ,Nicholson AM,Jenkins N,Knowles CJ

    更新日期:1991-12-05 00:00:00

  • Substrate inhibition kinetics for toluene and benzene degrading pure cultures and a method for collection and analysis of respirometric data for strongly inhibited cultures.

    abstract::We present an evaluation of the qualitative and quantitative effects that high concentrations of benzene and toluene have on the growth rate of several pure cultures that use these compounds as their sole carbon and energy source. The cultures employed were five widely studied environmental isolates: Pseudomonas putid...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.10729

    authors: Alagappan G,Cowan R

    更新日期:2003-09-30 00:00:00

  • Conversion of L-sorbose to L-sorbosone by immobilized cells of Gluconobacter melanogenus IFO 3293.

    abstract::Gluconobacter melanogenus IFO 3293 cells capable of converting L-sorbose to L-sorbosone were immobilized in polyacrylamide gel. The preferred polymer composition for high activity and stability was determined to contain a total monomer concentration of 7.2% and 16.6% crosslinking agent. No significant differences in o...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260180208

    authors: Martin CK,Perlman D

    更新日期:1976-02-01 00:00:00

  • Stopped-flow kinetic studies of the cellulase-catalyzed conversion of cellulose to glucose.

    abstract::The kinetics of the cellulase-catalyzed conversion of soluble cellulose into glucose have been studied over a range of substrate concentrations and temperatures, and at pH values ranging from 4.75 to 7.0. Lineweaver-Burk plots were linear and led to V = 6.2muM/s and K(m) = 13.1 mM at pH 5.8 and 25.0 degrees C. The pK ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260251011

    authors: Banerjee DK,Laidler KJ

    更新日期:1983-10-01 00:00:00

  • Computer aided analysis for biosensing and screening.

    abstract::Biosensors with animal and microbial cells immobilized close to the tip of a membrane electrode have been developed for chemical and drug testing. Our experimental results show that biosensors can be used for drug screening and to provide useful information about various cell-chemical interactions. A computer aided an...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260310311

    authors: Li XM,Liang BS,Wang HY

    更新日期:1988-02-20 00:00:00

  • Itaconic acid production from glycerol using Escherichia coli harboring a random synonymous codon-substituted 5'-coding region variant of the cadA gene.

    abstract::Aspergillus terreus cadA, encoding cis-aconitate decarboxylase, is an essential gene for itaconic acid (IA) biosynthesis, but it is primarily expressed as insoluble aggregates in most industrial hosts. This has been a hurdle for the development of recombinant strategies for IA production. Here, we created a library of...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.25914

    authors: Jeon HG,Cheong DE,Han Y,Song JJ,Choi JH

    更新日期:2016-07-01 00:00:00

  • High concentrations of clavulanic acid but not of its degradation products decrease glycerol consumption and oxygen uptake rates in cultures of Streptomyces clavuligerus.

    abstract::Product inhibition of biological production systems is a widely observed phenomenon with prominent implications for the design and ultimately the success of biotechnological processes. In order to investigate whether such effects could limit the maximal concentrations in the production of the beta-lactamase inhibitor ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.21783

    authors: Brethauer S,Held M,Panke S

    更新日期:2008-06-15 00:00:00

  • Multivariable control of alcohol concentrations in the production of polyhydroxyalkanoates (PHAs) by Paracoccus denitrificans.

    abstract::A novel multivariable control strategy is developed for alcohol (ethanol and n-pentanol) concentrations in the production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate), P(HB-co-HV), a biodegradable polymer by Paracoccus denitrificans ATCC 1774. This controller, which is developed to control the mole fraction of P(HB...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.1101

    authors: Chanprateep S,Abe N,Shimizu H,Yamane T,Shioya S

    更新日期:2001-07-20 00:00:00

  • Application of high throughput pretreatment and co-hydrolysis system to thermochemical pretreatment. Part 2: Dilute alkali.

    abstract::High throughput pretreatment (HTPH) and enzymatic hydrolysis systems are now vital for screening large numbers of biomass samples to investigate biomass recalcitrance over various pretreatment and enzymatic hydrolysis conditions. Although hydrothermal pretreatment is currently being employed in most high throughput ap...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.24951

    authors: Li H,Gao X,Demartini JD,Kumar R,Wyman CE

    更新日期:2013-11-01 00:00:00

  • Influence of mass transfer limitations on determination of the half saturation constant for hydrogen uptake in a mixed-culture CH(4)-producing enrichment.

    abstract::There is strong evidence in the literature supporting the existence of significant mass transfer limitations on the kinetics of exogenous H(2) consumption by methanogens. The half saturation constant for H (2) uptake by a mixed-culture, CH(4) producing enrichment was measured using an experimental protocol that avoide...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260400704

    authors: Giraldo-Gomez E,Goodwin S,Switzenbaum MS

    更新日期:1992-10-05 00:00:00

  • Immobilization of Candida rugosa lipase on colloidal gas aphrons (CGAs).

    abstract::A novel technique for immobilization of Candida rugosa lipase onto anionic colloidal gas aphrons (CGAs) is described. CGAs are spherical microbubbles (10-100 microm) composed of an inner gas core surrounded by a surfactant shell. In this initial study, greater than 80% lipase (w/w) was effectively retained on the CGAs...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.1116

    authors: O'Connell PJ,Varley J

    更新日期:2001-08-05 00:00:00

  • Measurement of heat evolution and correlation with oxygen consumption during microbial growth. Reprinted from Biotechnology and Bioengineering, Vol. XI, Issue 3, Pages 269-281 (1968).

    abstract::A procedure for measuring the rate of heat production from a fermentation has been developed. The method is based on measuring the rate of temperature rise of the fermentation broth resulting from metabolism, when the temperature controller is turned off. The heat accumulation measured in this manner is then corrected...

    journal_title:Biotechnology and bioengineering

    pub_type: 传,古典文章,历史文章,杂志文章

    doi:10.1002/(sici)1097-0290(20000320)67:6<691::aid-bit

    authors: Cooney CL,Wang DI,Mateles RI

    更新日期:2000-03-20 00:00:00

  • Effect of matrices on affinity purification of protein C.

    abstract::One of the critical problems in scale-up of affinity chromatography is the mechanical strength of the support matrix against pressure. Because the costs of both the gel matrix and the ligand for the affinity chromatography are very high, the reusability of gel matrices is directly related to the total production cost....

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260391104

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

    更新日期:1992-05-01 00:00:00

  • Coculture of genetically transformed roots and shoots for synthesis, translocation, and biotransformation of secondary metabolites.

    abstract::Genetically transformed shooty teratomas of Atropa belladonna and a Duboisia leichhardtii x D. myoporoides hybrid were studied for biotransformation of tropane alkaloids in shake flasks and bioreactors. Although de novo synthesis of hyoscyamine and scopolamine was limited, shoots of both species were able to transloca...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/(SICI)1097-0290(19960305)49:5<481::AID-BIT

    authors: Subroto MA,Kwok KH,Hamill JD,Doran PM

    更新日期:1996-03-05 00:00:00