Rational design of allosteric regulation of homoserine dehydrogenase by a nonnatural inhibitor L-lysine.

Abstract:

:Allosteric proteins, which can sense different signals, are interesting biological parts for synthetic biology. In particular, the design of an artificial allosteric enzyme to sense an unnatural signal is both challenging and highly desired, for example, for a precise and dynamical control of fluxes of growth-essential but byproduct pathways in metabolic engineering of industrial microorganisms. In this work, we used homoserine dehydrogenase (HSDH) of Corynebacterium glutamicum, which is naturally allosterically regulated by threonine and isoleucine, as an example to demonstrate the feasibility of reengineering an allosteric enzyme to respond to an unnatural inhibitor L-lysine. For this purpose, the natural threonine binding sites of HSD were first predicted and verified by mutagenesis experiments. The threonine binding sites were then engineered to a lysine binding pocket. The reengineered HSD only responds to lysine inhibition but not to threonine. This is a significant step toward the construction of artificial molecular circuits for dynamic control of growth-essential byproduct formation pathway for lysine biosynthesis.

journal_name

ACS Synth Biol

journal_title

ACS synthetic biology

authors

Chen Z,Rappert S,Zeng AP

doi

10.1021/sb400133g

subject

Has Abstract

pub_date

2015-02-20 00:00:00

pages

126-31

issue

2

issn

2161-5063

journal_volume

4

pub_type

杂志文章
  • 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

  • A Single Cas9-VPR Nuclease for Simultaneous Gene Activation, Repression, and Editing in Saccharomyces cerevisiae.

    abstract::Combinatorial metabolic engineering has been widely established for the development of efficient microbial cell factories to produce the products of interest by precisely regulating the expression levels of multiple genes simultaneously. Here, we report a novel multifunctional CRISPR system that enables simultaneous g...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.0c00218

    authors: Dong C,Jiang L,Xu S,Huang L,Cai J,Lian J,Xu Z

    更新日期:2020-09-18 00:00:00

  • Lessons from Two Design-Build-Test-Learn Cycles of Dodecanol Production in Escherichia coli Aided by Machine Learning.

    abstract::The Design-Build-Test-Learn (DBTL) cycle, facilitated by exponentially improving capabilities in synthetic biology, is an increasingly adopted metabolic engineering framework that represents a more systematic and efficient approach to strain development than historical efforts in biofuels and biobased products. Here, ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.9b00020

    authors: Opgenorth P,Costello Z,Okada T,Goyal G,Chen Y,Gin J,Benites V,de Raad M,Northen TR,Deng K,Deutsch S,Baidoo EEK,Petzold CJ,Hillson NJ,Garcia Martin H,Beller HR

    更新日期:2019-06-21 00:00:00

  • An Orthogonal Multi-input Integration System to Control Gene Expression in Escherichia coli.

    abstract::In many biotechnological applications, it is useful for gene expression to be regulated by multiple signals, as this allows the programming of complex behavior. Here we implement, in Escherichia coli, a system that compares the concentration of two signal molecules, and tunes GFP expression proportionally to their rel...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.7b00109

    authors: Annunziata F,Matyjaszkiewicz A,Fiore G,Grierson CS,Marucci L,di Bernardo M,Savery NJ

    更新日期:2017-10-20 00:00:00

  • Cellular signaling circuits interfaced with synthetic, post-translational, negating Boolean logic devices.

    abstract::A negating functionality is fundamental to information processing of logic circuits within cells and computers. Aiming to adapt unutilized electronic concepts to the interrogation of signaling circuits in cells, we first took a bottom-up strategy whereby we created protein-based devices that perform negating Boolean l...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb500222z

    authors: Razavi S,Su S,Inoue T

    更新日期:2014-09-19 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

  • Exploring the Nonconserved Sequence Space of Synthetic Expression Modules in Bacillus subtilis.

    abstract::Increasing protein expression levels is a key step in the commercial production of enzymes. Predicting promoter activity and translation initiation efficiency based solely on consensus sequences have so far met with mixed results. Here, we addressed this challenge using a "brute-force" approach by designing and synthe...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00110

    authors: Sauer C,Ver Loren van Themaat E,Boender LGM,Groothuis D,Cruz R,Hamoen LW,Harwood CR,van Rij T

    更新日期:2018-07-20 00:00:00

  • Effects of compartment size on the kinetics of intracompartmental multimeric protein synthesis.

    abstract::The cell contents are encapsulated within a compartment, the volume of which is a fundamental physical parameter that may affect intracompartmental reactions. However, there have been few studies to elucidate whether and how volume changes alone can affect the reaction kinetics. It is difficult to address these questi...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb300041z

    authors: Matsuura T,Hosoda K,Kazuta Y,Ichihashi N,Suzuki H,Yomo T

    更新日期:2012-09-21 00:00:00

  • Mathematical Modeling of RNA-Based Architectures for Closed Loop Control of Gene Expression.

    abstract::Feedback allows biological systems to control gene expression precisely and reliably, even in the presence of uncertainty, by sensing and processing environmental changes. Taking inspiration from natural architectures, synthetic biologists have engineered feedback loops to tune the dynamics and improve the robustness ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00040

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

    更新日期:2018-05-18 00:00:00

  • Active Analyte Import Improves the Dynamic Range and Sensitivity of a Vitamin B12 Biosensor.

    abstract::Cell-free systems provide a versatile platform for the development of low-cost, easy-to-use sensors for diverse analytes. However, sensor affinity dictates response sensitivity, and improving binding affinity can be challenging. Here, we describe efforts to address this problem while developing a biosensor for vitamin...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.9b00429

    authors: McNerney MP,Piorino F,Michel CL,Styczynski MP

    更新日期:2020-02-21 00:00:00

  • Genome Engineering of Virulent Lactococcal Phages Using CRISPR-Cas9.

    abstract::Phages are biological entities found in every ecosystem. Although much has been learned about them in past decades, significant knowledge gaps remain. Manipulating virulent phage genomes is challenging. To date, no efficient gene-editing tools exist for engineering virulent lactococcal phages. Lactococcus lactis is a ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.6b00388

    authors: Lemay ML,Tremblay DM,Moineau S

    更新日期:2017-07-21 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

  • Microbial Synthesis of Human-Hormone Melatonin at Gram Scales.

    abstract::Melatonin is a commercially attractive tryptophan-derived hormone. Here we describe a bioprocess for the production of melatonin using Escherichia coli to high titers. The first engineered strain produced 0.13 g/L of melatonin from tryptophan under fed-batch fermentation conditions. A 4-fold improvement on melatonin t...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.0c00065

    authors: Luo H,Schneider K,Christensen U,Lei Y,Herrgard M,Palsson BØ

    更新日期:2020-06-19 00:00:00

  • Engineering Escherichia coli into a protein delivery system for mammalian cells.

    abstract::Many Gram-negative pathogens encode type 3 secretion systems, sophisticated nanomachines that deliver proteins directly into the cytoplasm of mammalian cells. These systems present attractive opportunities for therapeutic protein delivery applications; however, their utility has been limited by their inherent pathogen...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.5b00002

    authors: Reeves AZ,Spears WE,Du J,Tan KY,Wagers AJ,Lesser CF

    更新日期:2015-05-15 00:00:00

  • 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

  • Synthetic Biogenesis of Bacterial Amyloid Nanomaterials with Tunable Inorganic-Organic Interfaces and Electrical Conductivity.

    abstract::Amyloids are highly ordered, hierarchal protein nanoassemblies. Functional amyloids in bacterial biofilms, such as Escherichia coli curli fibers, are formed by the polymerization of monomeric proteins secreted into the extracellular space. Curli is synthesized by living cells, is primarily composed of the major curlin...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.6b00166

    authors: Seker UO,Chen AY,Citorik RJ,Lu TK

    更新日期:2017-02-17 00:00:00

  • Conditional Recruitment to a DNA-Bound CRISPR-Cas Complex Using a Colocalization-Dependent Protein Switch.

    abstract::To spatially control biochemical functions at specific sites within a genome, we have engineered a synthetic switch that activates when bound to its DNA target site. The system uses two CRISPR-Cas complexes to colocalize components of a de novo-designed protein switch (Co-LOCKR) to adjacent sites in the genome. Coloca...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.0c00012

    authors: Kirkpatrick RL,Lewis K,Langan RA,Lajoie MJ,Boyken SE,Eakman M,Baker D,Zalatan JG

    更新日期:2020-09-18 00:00:00

  • A Genetically Encoded Protein Polymer for Uranyl Binding and Extraction Based on the SpyTag-SpyCatcher Chemistry.

    abstract::A defining goal of synthetic biology is to develop biomaterials with superior performance and versatility. Here we introduce a purely genetically encoded and self-assembling biopolymer based on the SpyTag-SpyCatcher chemistry. We show the application of this polymer for highly efficient uranyl binding and extraction f...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00223

    authors: Yang X,Wei J,Wang Y,Yang C,Zhao S,Li C,Dong Y,Bai K,Li Y,Teng H,Wang D,Lyu N,Li J,Chang X,Ning X,Ouyang Q,Zhang Y,Qian L

    更新日期:2018-10-19 00:00:00

  • Manipulation of endogenous kinase activity in living cells using photoswitchable inhibitory peptides.

    abstract::Optogenetic control of endogenous signaling can be an important tool for probing cell behavior. Using the photoresponse of the LOV2 domain of Avena sativa phototropin 1, we developed analogues of kinase inhibitors whose activity is light dependent. Inhibitory peptides were appended to the Jα helix, where they potently...

    journal_title:ACS synthetic biology

    pub_type: 信件

    doi:10.1021/sb5001356

    authors: Yi JJ,Wang H,Vilela M,Danuser G,Hahn KM

    更新日期:2014-11-21 00:00:00

  • PERSIA for Direct Fluorescence Measurements of Transcription, Translation, and Enzyme Activity in Cell-Free Systems.

    abstract::Quantification of biology's central dogma (transcription and translation) is pursued by a variety of methods. Direct, immediate, and ongoing quantification of these events is difficult to achieve. Common practice is to use fluorescent or luminescent proteins to report indirectly on prior cellular events, such as turni...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00450

    authors: Wick S,Walsh DI 3rd,Bobrow J,Hamad-Schifferli K,Kong DS,Thorsen T,Mroszczyk K,Carr PA

    更新日期:2019-05-17 00:00:00

  • Robust Characterization of Two Distinct Glutarate Sensing Transcription Factors of Pseudomonas putida l-Lysine Metabolism.

    abstract::A significant bottleneck in synthetic biology involves screening large genetically encoded libraries for desirable phenotypes such as chemical production. However, transcription factor-based biosensors can be leveraged to screen thousands of genetic designs for optimal chemical production in engineered microbes. In th...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.9b00255

    authors: Thompson MG,Costello Z,Hummel NFC,Cruz-Morales P,Blake-Hedges JM,Krishna RN,Skyrud W,Pearson AN,Incha MR,Shih PM,Garcia-Martin H,Keasling JD

    更新日期:2019-10-18 00:00:00

  • A Post-translational Metabolic Switch Enables Complete Decoupling of Bacterial Growth from Biopolymer Production in Engineered Escherichia coli.

    abstract::Most of the current methods for controlling the formation rate of a key protein or enzyme in cell factories rely on the manipulation of target genes within the pathway. In this article, we present a novel synthetic system for post-translational regulation of protein levels, FENIX, which provides both independent contr...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00345

    authors: Durante-Rodríguez G,de Lorenzo V,Nikel PI

    更新日期:2018-11-16 00:00:00

  • Engineering the S-Layer of Caulobacter crescentus as a Foundation for Stable, High-Density, 2D Living Materials.

    abstract::Materials synthesized by organisms, such as bones and wood, combine the ability to self-repair with remarkable mechanical properties. This multifunctionality arises from the presence of living cells within the material and hierarchical assembly of different components across nanometer to micron scales. While creating ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00448

    authors: Charrier M,Li D,Mann VR,Yun L,Jani S,Rad B,Cohen BE,Ashby PD,Ryan KR,Ajo-Franklin CM

    更新日期:2019-01-18 00:00:00

  • Tn-Core: A Toolbox for Integrating Tn-seq Gene Essentiality Data and Constraint-Based Metabolic Modeling.

    abstract::The design of synthetic cells requires a detailed understanding of the relevance of genes and gene networks underlying complex cellular phenotypes. Transposon-sequencing (Tn-seq) and constraint-based metabolic modeling can be used to probe the core genetic and metabolic networks underlying a biological process. Integr...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00432

    authors: diCenzo GC,Mengoni A,Fondi M

    更新日期:2019-01-18 00:00:00

  • Rapid and Scalable Preparation of Bacterial Lysates for Cell-Free Gene Expression.

    abstract::Cell-free gene expression systems are emerging as an important platform for a diverse range of synthetic biology and biotechnology applications, including production of robust field-ready biosensors. Here, we combine programmed cellular autolysis with a freeze-thaw or freeze-dry cycle to create a practical, reproducib...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.7b00253

    authors: Didovyk A,Tonooka T,Tsimring L,Hasty J

    更新日期:2017-12-15 00:00:00

  • SBOLExplorer: Data Infrastructure and Data Mining for Genetic Design Repositories.

    abstract::This paper describes SBOLExplorer, a system that is used to provide intuitive searching within the SynBioHub genetic design repository. SynBioHub stores genetic constructs encoded in the SBOL data format. These constructs can represent genetic parts, circuits, and sequences. These constructs are often numerous, exist ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.9b00089

    authors: Zhang M,Zundel Z,Myers CJ

    更新日期:2019-10-18 00:00:00

  • Matching Protein Interfaces for Improved Medium-Chain Fatty Acid Production.

    abstract::Medium-chain fatty acids (MCFAs) are key intermediates in the synthesis of medium-chain chemicals including α-olefins and dicarboxylic acids. In bacteria, microbial production of MCFAs is limited by the activity and product profile of fatty acyl-ACP thioesterases. Here, we engineer a heterologous bacterial medium-chai...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.7b00334

    authors: Sarria S,Bartholow TG,Verga A,Burkart MD,Peralta-Yahya P

    更新日期:2018-05-18 00:00:00

  • Fluorescent proteins and in vitro genetic organization for cell-free synthetic biology.

    abstract::To facilitate the construction of cell-free genetic devices, we evaluated the ability of 17 different fluorescent proteins to give easily detectable fluorescence signals in real-time from in vitro transcription-translation reactions with a minimal system consisting of T7 RNA polymerase and E. coli translation machiner...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb400003y

    authors: Lentini R,Forlin M,Martini L,Del Bianco C,Spencer AC,Torino D,Mansy SS

    更新日期:2013-09-20 00:00:00

  • Synthetic Derivatives of (+)-epi-α-Bisabolol Are Formed by Mammalian Cytochromes P450 Expressed in a Yeast Reconstituted Pathway.

    abstract::Identification of the enzyme(s) involved in complex biosynthetic pathways can be challenging. An alternative approach might be to deliberately diverge from the original natural enzyme source and use promiscuous enzymes from other organisms. In this paper, we have tested the ability of a series of human and animal cyto...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.9b00399

    authors: Sarrade-Loucheur A,Ro DK,Fauré R,Remaud-Siméon M,Truan G

    更新日期:2020-02-21 00:00:00

  • Metabolic Engineering of Escherichia coli for Efficient Production of 2-Pyrone-4,6-dicarboxylic Acid from Glucose.

    abstract::2-Pyrone-4,6-dicarboxylic acid (PDC) is a pseudoaromatic dicarboxylic acid and is a promising biobased building block chemical that can be used to make diverse polyesters with novel functionalities. In this study, Escherichia coli was metabolically engineered to produce PDC from glucose. First, an efficient biosynthet...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00281

    authors: Luo ZW,Kim WJ,Lee SY

    更新日期:2018-09-21 00:00:00