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Synthetic Biology Toolbox for Antarctic Pseudomonas sp. Strains: Toward a Psychrophilic Nonmodel Chassis for Function-Driven Metagenomics.
ACS Synth Biol. 2023 03 17; 12(3):722-734.AS

Abstract

One major limitation of function-driven metagenomics is the ability of the host to express the metagenomic DNA correctly. Differences in the transcriptional, translational, and post-translational machinery between the organism to which the DNA belongs and the host strain are all factors that influence the success of a functional screening. For this reason, the use of alternative hosts is an appropriate approach to favor the identification of enzymatic activities in function-driven metagenomics. To be implemented, appropriate tools should be designed to build the metagenomic libraries in those hosts. Moreover, discovery of new chassis and characterization of synthetic biology toolbox in nonmodel bacteria is an active field of research to expand the potential of these organisms in processes of industrial interest. Here, we assessed the suitability of two Antarctic psychrotolerant Pseudomonas strains as putative alternative hosts for function-driven metagenomics using pSEVA modular vectors as scaffold. We determined a set of synthetic biology tools suitable for these hosts and, as a proof of concept, we demonstrated their fitness for heterologous protein expression. These hosts represent a step forward for the prospection and identification of psychrophilic enzymes of biotechnological interest.

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Authors+Show Affiliations

Amarelle V
Departamento de Bioquímica y Genómica Microbianas. Instituto de Investigaciones Biológicas Clemente Estable, Av. Italia 3318, Montevideo 11600, Uruguay.
Roldán DM
Departamento de Bioquímica y Genómica Microbianas. Instituto de Investigaciones Biológicas Clemente Estable, Av. Italia 3318, Montevideo 11600, Uruguay.
Fabiano E
Departamento de Bioquímica y Genómica Microbianas. Instituto de Investigaciones Biológicas Clemente Estable, Av. Italia 3318, Montevideo 11600, Uruguay.
Guazzaroni ME
Departamento de Biologia. FFCLRP, University of São Paulo, 14049-901 Ribeirão Preto, São Paulo, Brazil.

MeSH

Synthetic BiologyPseudomonasMetagenomicsAntarctic RegionsBiotechnology

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

36862944

Citation

Amarelle, Vanesa, et al. "Synthetic Biology Toolbox for Antarctic Pseudomonas Sp. Strains: Toward a Psychrophilic Nonmodel Chassis for Function-Driven Metagenomics." ACS Synthetic Biology, vol. 12, no. 3, 2023, pp. 722-734.
Amarelle V, Roldán DM, Fabiano E, et al. Synthetic Biology Toolbox for Antarctic Pseudomonas sp. Strains: Toward a Psychrophilic Nonmodel Chassis for Function-Driven Metagenomics. ACS Synth Biol. 2023;12(3):722-734.
Amarelle, V., Roldán, D. M., Fabiano, E., & Guazzaroni, M. E. (2023). Synthetic Biology Toolbox for Antarctic Pseudomonas sp. Strains: Toward a Psychrophilic Nonmodel Chassis for Function-Driven Metagenomics. ACS Synthetic Biology, 12(3), 722-734. https://doi.org/10.1021/acssynbio.2c00543
Amarelle V, et al. Synthetic Biology Toolbox for Antarctic Pseudomonas Sp. Strains: Toward a Psychrophilic Nonmodel Chassis for Function-Driven Metagenomics. ACS Synth Biol. 2023 03 17;12(3):722-734. PubMed PMID: 36862944.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Synthetic Biology Toolbox for Antarctic Pseudomonas sp. Strains: Toward a Psychrophilic Nonmodel Chassis for Function-Driven Metagenomics. AU - Amarelle,Vanesa, AU - Roldán,Diego M, AU - Fabiano,Elena, AU - Guazzaroni,María-Eugenia, Y1 - 2023/03/02/ PY - 2023/3/3/pubmed PY - 2023/3/21/medline PY - 2023/3/2/entrez KW - functional metagenomics KW - nonmodel chassis KW - synthetic biology toolbox SP - 722 EP - 734 JF - ACS synthetic biology JO - ACS Synth Biol VL - 12 IS - 3 N2 - One major limitation of function-driven metagenomics is the ability of the host to express the metagenomic DNA correctly. Differences in the transcriptional, translational, and post-translational machinery between the organism to which the DNA belongs and the host strain are all factors that influence the success of a functional screening. For this reason, the use of alternative hosts is an appropriate approach to favor the identification of enzymatic activities in function-driven metagenomics. To be implemented, appropriate tools should be designed to build the metagenomic libraries in those hosts. Moreover, discovery of new chassis and characterization of synthetic biology toolbox in nonmodel bacteria is an active field of research to expand the potential of these organisms in processes of industrial interest. Here, we assessed the suitability of two Antarctic psychrotolerant Pseudomonas strains as putative alternative hosts for function-driven metagenomics using pSEVA modular vectors as scaffold. We determined a set of synthetic biology tools suitable for these hosts and, as a proof of concept, we demonstrated their fitness for heterologous protein expression. These hosts represent a step forward for the prospection and identification of psychrophilic enzymes of biotechnological interest. SN - 2161-5063 UR - https://www.unboundmedicine.com/medline/citation/36862944/Synthetic_Biology_Toolbox_for_Antarctic_Pseudomonas_sp__Strains:_Toward_a_Psychrophilic_Nonmodel_Chassis_for_Function_Driven_Metagenomics_ DB - PRIME DP - Unbound Medicine ER -