Identification of a Strong Quorum Sensing- and Thermo-Regulated Promoter for the Biosynthesis of a New Metabolite Pesticide Phenazine-1-carboxamide in Pseudomonas strain PA1201.

Abstract:

:Phenazine-1-carboxamide (PCN) produced by multifarious Pseudomonas strains represents a promising candidate as a new metabolite pesticide due to its broad-spectrum antifungal activity and capacity to induce systemic resistance in plants. The rice rhizosphere Pseudomonas strain PA1201 contains two reiterated gene clusters, phz1 and phz2, for phenazine-1-carboxylic acid (PCA) biosynthesis; PCA is further converted into PCN by this strain using a functional phzH-encoding glutamine aminotransferase. However, PCN levels in PA1201 constitute approximately one-fifth of PCA levels and the optimal temperature for PCN synthesis is 28 °C. In this study, the phzH open reading frame (ORF) and promoter region were investigated and reannotated. phzH promoter PphzH was found to be a weak promoter, and PhzH levels were not sufficient to convert all of the native PCA into PCN. Following RNA Seq and promoter-lacZ fusion analyses, a strong quorum sensing (QS)- and thermo-regulated promoter PrhlI was identified and characterized. The activity of PphzH is approximately 1% of PrhlI in PA1201. After three rounds of promoter editing and swapping by PrhlI, a new PCN-overproducing strain UP46 was generated. The optimal fermentation temperature for PCN biosynthesis in UP46 was increased from 28 to 37 °C and the PCN fermentation titer increased 179.5-fold, reaching 14.1 g/L, the highest ever reported.

journal_name

ACS Synth Biol

journal_title

ACS synthetic biology

authors

Jin ZJ,Zhou L,Sun S,Cui Y,Song K,Zhang X,He YW

doi

10.1021/acssynbio.0c00161

subject

Has Abstract

pub_date

2020-07-17 00:00:00

pages

1802-1812

issue

7

issn

2161-5063

journal_volume

9

pub_type

杂志文章
  • A Novel Tool for Microbial Genome Editing Using the Restriction-Modification System.

    abstract::Scarless genetic manipulation of genomes is an essential tool for biological research. The restriction-modification (R-M) system is a defense system in bacteria that protects against invading genomes on the basis of its ability to distinguish foreign DNA from self DNA. Here, we designed an R-M system-mediated genome e...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.7b00254

    authors: Bai H,Deng A,Liu S,Cui D,Qiu Q,Wang L,Yang Z,Wu J,Shang X,Zhang Y,Wen T

    更新日期:2018-01-19 00:00:00

  • Comprehensive Profiling of Four Base Overhang Ligation Fidelity by T4 DNA Ligase and Application to DNA Assembly.

    abstract::Synthetic biology relies on the manufacture of large and complex DNA constructs from libraries of genetic parts. Golden Gate and other Type IIS restriction enzyme-dependent DNA assembly methods enable rapid construction of genes and operons through one-pot, multifragment assembly, with the ordering of parts determined...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00333

    authors: Potapov V,Ong JL,Kucera RB,Langhorst BW,Bilotti K,Pryor JM,Cantor EJ,Canton B,Knight TF,Evans TC Jr,Lohman GJS

    更新日期:2018-11-16 00:00:00

  • Multiple T-DNA Delivery to Plants Using Novel Mini Binary Vectors with Compatible Replication Origins.

    abstract::Improved plants are necessary to meet human needs. Agrobacterium-mediated transformation is the most common method used to rewire plant capabilities. For plant gene delivery, DNA constructs are assembled into binary T-DNA vectors that rely on broad host range origins for bacterial replication. Here we present pLX vect...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.6b00354

    authors: Pasin F,Bedoya LC,Bernabé-Orts JM,Gallo A,Simón-Mateo C,Orzaez D,García JA

    更新日期:2017-10-20 00:00:00

  • Reconstituted biosynthesis of the nonribosomal macrolactone antibiotic valinomycin in Escherichia coli.

    abstract::The structural complexity of nonribosomal peptides (NRPs) impeding economic chemical synthesis and poor cultivability of source organisms limits the development of bioprocesses for novel bioactive compounds. Since nonribosomal peptide synthetases (NRPSs) assemble NRPs from simple amino acid building blocks, heterologo...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb400082j

    authors: Jaitzig J,Li J,Süssmuth RD,Neubauer P

    更新日期:2014-07-18 00:00:00

  • A Scalable Epitope Tagging Approach for High Throughput ChIP-Seq Analysis.

    abstract::Eukaryotic transcriptional factors (TFs) typically recognize short genomic sequences alone or together with other proteins to modulate gene expression. Mapping of TF-DNA interactions in the genome is crucial for understanding the gene regulatory programs in cells. While chromatin immunoprecipitation followed by sequen...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.6b00358

    authors: Xiong X,Zhang Y,Yan J,Jain S,Chee S,Ren B,Zhao H

    更新日期:2017-06-16 00:00:00

  • A System for Gene Expression Noise Control in Yeast.

    abstract::Gene expression noise arises from stochastic variation in the synthesis and degradation of mRNA and protein molecules and creates differences in protein numbers across populations of genetically identical cells. Such variability can lead to imprecision and reduced performance of both native and synthetic networks. In ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00279

    authors: Mundt M,Anders A,Murray SM,Sourjik V

    更新日期:2018-11-16 00:00:00

  • RNA-Catalyzed Polymerization of Deoxyribose, Threose, and Arabinose Nucleic Acids.

    abstract::An RNA-dependent RNA polymerase ribozyme that was highly optimized through in vitro evolution for the ability to copy a broad range of template sequences exhibits promiscuity toward other nucleic acids and nucleic acid analogues, including DNA, threose nucleic acid (TNA), and arabinose nucleic acid (ANA). By operating...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.9b00044

    authors: Horning DP,Bala S,Chaput JC,Joyce GF

    更新日期:2019-05-17 00:00:00

  • Repurposing Synechocystis PCC6803 UirS-UirR as a UV-Violet/Green Photoreversible Transcriptional Regulatory Tool in E. coli.

    abstract::We have previously engineered green/red and red/far red photoreversible E. coli phytochrome and cyanobacteriochrome (CBCR) two-component systems (TCSs) and utilized them to program tailor-made gene expression signals for gene circuit characterization. Here, we transport the UV-violet/green photoreversible CBCR TCS Uir...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.6b00068

    authors: Ramakrishnan P,Tabor JJ

    更新日期:2016-07-15 00:00:00

  • Combined Assembly and Targeted Integration of Multigene for Nitrogenase Biosynthetic Pathway in Saccharomyces cerevisiae.

    abstract::Biological nitrogen fixation, a process unique to diazotrophic prokaryote, is catalyzed by the nitrogenase complex. There has been a long-standing interest in reconstituting a nitrogenase biosynthetic pathway in a eukaryotic host with the final aim of developing N2-fixing cereal crops. In this study, we report that a ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.9b00060

    authors: Liu X,Wang M,Song Y,Li Y,Liu P,Shi H,Li Y,Hao T,Zhang H,Jiang W,Chen S,Li J

    更新日期:2019-08-16 00:00:00

  • Synthetic tunable amplifying buffer circuit in E. coli.

    abstract::While predictable design of a genetic circuit's output is a major goal of synthetic biology, it remains a significant challenge because DNA binding sites in the cell affect the concentration of available transcription factors (TF). To mitigate this problem, we propose to use a TF that results from the (reversible) pho...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb5002533

    authors: Nilgiriwala KS,Jiménez J,Rivera PM,Del Vecchio D

    更新日期:2015-05-15 00:00:00

  • Incorporation of a Synthetic Amino Acid into dCas9 Improves Control of Gene Silencing.

    abstract::The CRISPR-Cas9 nuclease has been repurposed as a tool for gene repression (CRISPRi). This catalytically dead Cas9 (dCas9) variant inhibits transcription by blocking either initiation or elongation by the RNA polymerase complex. Conditional control of dCas9-mediated repression has been achieved with inducible promoter...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00347

    authors: Koopal B,Kruis AJ,Claassens NJ,Nobrega FL,van der Oost J

    更新日期:2019-02-15 00:00:00

  • Nucleic Acid Detection Using CRISPR/Cas Biosensing Technologies.

    abstract::For infectious diseases, rapid and accurate identification of the pathogen is critical for effective management and treatment, but diagnosis remains challenging, particularly in resource-limited areas. Methods that accurately detect pathogen nucleic acids can provide robust, accurate, rapid, and ultrasensitive technol...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.9b00507

    authors: Aman R,Mahas A,Mahfouz M

    更新日期:2020-06-19 00:00:00

  • Genetically Encodable Bacterial Flavin Transferase for Fluorogenic Protein Modification in Mammalian Cells.

    abstract::A bacterial flavin transferase (ApbE) was recently employed for flavin mononucleotide (FMN) modification on the Na+-translocating NADH:quinone oxidoreductase C (NqrC) protein in the pathogenic Gram-negative bacterium Vibrio cholerae. We employed this unique post-translational modification in mammalian cells and found ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.6b00284

    authors: Kang MG,Park J,Balboni G,Lim MH,Lee C,Rhee HW

    更新日期:2017-04-21 00:00:00

  • FlopR: An Open Source Software Package for Calibration and Normalization of Plate Reader and Flow Cytometry Data.

    abstract::The measurement of gene expression using fluorescence markers has been a cornerstone of synthetic biology for the past two decades. However, the use of arbitrary units has limited the usefulness of these data for many quantitative purposes. Calibration of fluorescence measurements from flow cytometry and plate reader ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.0c00296

    authors: Fedorec AJH,Robinson CM,Wen KY,Barnes CP

    更新日期:2020-09-18 00:00:00

  • The Experiment Data Depot: A Web-Based Software Tool for Biological Experimental Data Storage, Sharing, and Visualization.

    abstract::Although recent advances in synthetic biology allow us to produce biological designs more efficiently than ever, our ability to predict the end result of these designs is still nascent. Predictive models require large amounts of high-quality data to be parametrized and tested, which are not generally available. Here, ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.7b00204

    authors: Morrell WC,Birkel GW,Forrer M,Lopez T,Backman TWH,Dussault M,Petzold CJ,Baidoo EEK,Costello Z,Ando D,Alonso-Gutierrez J,George KW,Mukhopadhyay A,Vaino I,Keasling JD,Adams PD,Hillson NJ,Garcia Martin H

    更新日期:2017-12-15 00:00:00

  • Merlin: Computer-Aided Oligonucleotide Design for Large Scale Genome Engineering with MAGE.

    abstract::Genome engineering technologies now enable precise manipulation of organism genotype, but can be limited in scalability by their design requirements. Here we describe Merlin ( http://merlincad.org ), an open-source web-based tool to assist biologists in designing experiments using multiplex automated genome engineerin...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.5b00219

    authors: Quintin M,Ma NJ,Ahmed S,Bhatia S,Lewis A,Isaacs FJ,Densmore D

    更新日期:2016-06-17 00:00:00

  • A Tunable Protein Piston That Breaks Membranes to Release Encapsulated Cargo.

    abstract::Movement of molecules across membranes in response to a stimulus is a key component of cellular programming. Here, we characterize and manipulate the response of a protein-based piston capable of puncturing membranes in a pH-dependent manner. Our protein actuator consists of modified R bodies found in a bacterial endo...

    journal_title:ACS synthetic biology

    pub_type: 信件

    doi:10.1021/acssynbio.5b00237

    authors: Polka JK,Silver PA

    更新日期:2016-04-15 00:00:00

  • Strategies for Editing Virulent Staphylococcal Phages Using CRISPR-Cas10.

    abstract::Staphylococci are prevalent skin-dwelling bacteria that are also leading causes of antibiotic-resistant infections. Viruses that infect and lyse these organisms (virulent staphylococcal phages) can be used as alternatives to conventional antibiotics and represent promising tools to eliminate or manipulate specific spe...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.7b00240

    authors: Bari SMN,Walker FC,Cater K,Aslan B,Hatoum-Aslan A

    更新日期:2017-12-15 00:00:00

  • Direct mutagenesis of thousands of genomic targets using microarray-derived oligonucleotides.

    abstract::Multiplex Automated Genome Engineering (MAGE) allows simultaneous mutagenesis of multiple target sites in bacterial genomes using short oligonucleotides. However, large-scale mutagenesis requires hundreds to thousands of unique oligos, which are costly to synthesize and impossible to scale-up by traditional phosphoram...

    journal_title:ACS synthetic biology

    pub_type: 信件

    doi:10.1021/sb5001565

    authors: Bonde MT,Kosuri S,Genee HJ,Sarup-Lytzen K,Church GM,Sommer MO,Wang HH

    更新日期:2015-01-16 00:00:00

  • Assembly of Multicomponent Protein Filaments Using Engineered Subunit Interfaces.

    abstract::Exploiting the ability of proteins to self-assemble into architectural templates may provide novel routes for the positioning of functional molecules in nanotechnology. Here we report the engineering of multicomponent protein templates composed of distinct monomers that assemble in repeating orders into a dynamic func...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00241

    authors: Glover DJ,Lim S,Xu D,Sloan NB,Zhang Y,Clark DS

    更新日期:2018-10-19 00:00:00

  • Gene-Mediated Chemical Communication in Synthetic Protocell Communities.

    abstract::A gene-directed chemical communication pathway between synthetic protocell signaling transmitters (lipid vesicles) and receivers (proteinosomes) was designed, built and tested using a bottom-up modular approach comprising small molecule transcriptional control, cell-free gene expression, porin-directed efflux, substra...

    journal_title:ACS synthetic biology

    pub_type: 信件

    doi:10.1021/acssynbio.7b00306

    authors: Tang TD,Cecchi D,Fracasso G,Accardi D,Coutable-Pennarun A,Mansy SS,Perriman AW,Anderson JLR,Mann S

    更新日期:2018-02-16 00:00:00

  • Artificial conversion of the mating-type of Saccharomyces cerevisiae without autopolyploidization.

    abstract::Crossbreeding is a classical yeast hybridization procedure, where the mating of haploid cells of opposite mating-type, MATa and MATα cells, produces a new heterozygous diploid. Here, we describe a method to generate haploid MATa and MATα cells using mating-type conversion caused by expression of the HO gene, which enc...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb400016j

    authors: Fukuda N,Matsukura S,Honda S

    更新日期:2013-12-20 00:00:00

  • Analog Computation by DNA Strand Displacement Circuits.

    abstract::DNA circuits have been widely used to develop biological computing devices because of their high programmability and versatility. Here, we propose an architecture for the systematic construction of DNA circuits for analog computation based on DNA strand displacement. The elementary gates in our architecture include ad...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.6b00144

    authors: Song T,Garg S,Mokhtar R,Bui H,Reif J

    更新日期:2016-08-19 00:00:00

  • Switching Protein Localization by Site-Directed RNA Editing under Control of Light.

    abstract::Site directed RNA editing is an engineered tool for the posttranscriptional manipulation of RNA and proteins. Here, we demonstrate the inclusion of additional N- and C-terminal protein domains in an RNA editing-dependent manner to switch between protein isoforms in mammalian cell culture. By inclusion of localization ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.7b00113

    authors: Vogel P,Hanswillemenke A,Stafforst T

    更新日期:2017-09-15 00:00:00

  • The Fifth Annual Sc2.0 and Synthetic Genomes Conference: Synthetic Genomes in High Gear.

    abstract::The Sc2.0 project is perhaps the largest synthetic biology project in the public domain, and ultimately aims to construct a new version of the humble brewer's yeast, Saccharomyces cerevisiae. Each year, the Sc2.0 consortium gather to discuss progress in this ambitious project and highlight new developments at the fore...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章,评审

    doi:10.1021/acssynbio.6b00227

    authors: Walker RS,Cai Y

    更新日期:2016-09-16 00:00:00

  • Homologous Quorum Sensing Regulatory Circuit: A Dual-Input Genetic Controller for Modulating Quorum Sensing-Mediated Protein Expression in E. coli.

    abstract::We developed a hybrid synthetic circuit that co-opts the genetic regulation of the native bacterial quorum sensing autoinducer-2 and imposes an extra external controller for maintaining tightly controlled gene expression. This dual-input genetic controller was mathematically modeled and, by design, can be operated in ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.0c00179

    authors: Hauk P,Stephens K,Virgile C,VanArsdale E,Pottash AE,Schardt JS,Jay SM,Sintim HO,Bentley WE

    更新日期:2020-10-16 00:00:00

  • Metabolic Engineering of Pseudomonas putida KT2440 for Complete Mineralization of Methyl Parathion and γ-Hexachlorocyclohexane.

    abstract::Agricultural soils are often cocontaminated with multiple pesticides. Unfortunately, microorganisms isolated from natural environments do not possess the ability to simultaneously degrade different classes of pesticides. Currently, we can use the approaches of synthetic biology to create a strain endowed with various ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.6b00025

    authors: Gong T,Liu R,Zuo Z,Che Y,Yu H,Song C,Yang C

    更新日期:2016-05-20 00:00:00

  • Combinatory Biosynthesis of Prenylated 4-Hydroxybenzoate Derivatives by Overexpression of the Substrate-Promiscuous Prenyltransferase XimB in Engineered E. coli.

    abstract::Prenylated aromatic compounds are important intermediates in the biosynthesis of bioactive molecules such as 3-chromanols from plants, ubiquinones from prokaryotes and meroterpenoids from sponges. Biosynthesis of prenylated aromatic compounds using prokaryotic microorganisms has attracted increasing attention in the f...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00070

    authors: He BB,Bu XL,Zhou T,Li SM,Xu MJ,Xu J

    更新日期:2018-09-21 00:00:00

  • Rationalizing Context-Dependent Performance of Dynamic RNA Regulatory Devices.

    abstract::The ability of RNA to sense, regulate, and store information is an attractive attribute for a variety of functional applications including the development of regulatory control devices for synthetic biology. RNA folding and function is known to be highly context sensitive, which limits the modularity and reuse of RNA ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00041

    authors: Kent R,Halliwell S,Young K,Swainston N,Dixon N

    更新日期:2018-07-20 00:00:00

  • Development of an artificial calcium-dependent transcription factor to detect sustained intracellular calcium elevation.

    abstract::The development of a synthetic transcription factor that responds to intracellular calcium signals enables analyzing cellular events at the single-cell level or "rewiring" the intracellular information networks. In this study, we developed the calcium-dependent transcription factor (CaTF), which was cleaved by calpain...

    journal_title:ACS synthetic biology

    pub_type: 信件

    doi:10.1021/sb500070c

    authors: Suzuki S,Murotomi K,Nakajima Y,Kawai K,Ohta K,Warita K,Miki T,Takeuchi Y

    更新日期:2014-10-17 00:00:00