Cargando…

Application of serine integrases for secondary metabolite pathway assembly in Streptomyces

Serine integrases have been shown to be efficient tools for metabolic pathway assembly. To further improve the flexibility and efficiency of pathway engineering via serine integrases, we explored how multiple orthogonally active serine integrases can be applied for use in vitro for the heterologous...

Descripción completa

Detalles Bibliográficos
Autores principales: Gao, Hong, Taylor, Gabrielle, Evans, Stephanie K., Fogg, Paul C.M., Smith, Margaret C.M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: KeAi Publishing 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7306541/
https://www.ncbi.nlm.nih.gov/pubmed/32596521
http://dx.doi.org/10.1016/j.synbio.2020.05.006
_version_ 1783548674932998144
author Gao, Hong
Taylor, Gabrielle
Evans, Stephanie K.
Fogg, Paul C.M.
Smith, Margaret C.M.
author_facet Gao, Hong
Taylor, Gabrielle
Evans, Stephanie K.
Fogg, Paul C.M.
Smith, Margaret C.M.
author_sort Gao, Hong
collection PubMed
description Serine integrases have been shown to be efficient tools for metabolic pathway assembly. To further improve the flexibility and efficiency of pathway engineering via serine integrases, we explored how multiple orthogonally active serine integrases can be applied for use in vitro for the heterologous expression of complex biosynthesis pathways in Streptomyces spp., the major producers of useful bioactive natural products. The results show that multiple orthogonal serine integrases efficiently assemble the genes from a complex biosynthesis pathway in a single in vitro recombination reaction, potentially permitting a versatile combinatorial assembly approach. Furthermore, the assembly strategy also permitted the incorporation of a well-characterised promoter upstream of each gene for expression in a heterologous host. The results demonstrate how site-specific recombination based on orthogonal serine integrases can be applied in Streptomyces spp.
format Online
Article
Text
id pubmed-7306541
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher KeAi Publishing
record_format MEDLINE/PubMed
spelling pubmed-73065412020-06-25 Application of serine integrases for secondary metabolite pathway assembly in Streptomyces Gao, Hong Taylor, Gabrielle Evans, Stephanie K. Fogg, Paul C.M. Smith, Margaret C.M. Synth Syst Biotechnol Article Serine integrases have been shown to be efficient tools for metabolic pathway assembly. To further improve the flexibility and efficiency of pathway engineering via serine integrases, we explored how multiple orthogonally active serine integrases can be applied for use in vitro for the heterologous expression of complex biosynthesis pathways in Streptomyces spp., the major producers of useful bioactive natural products. The results show that multiple orthogonal serine integrases efficiently assemble the genes from a complex biosynthesis pathway in a single in vitro recombination reaction, potentially permitting a versatile combinatorial assembly approach. Furthermore, the assembly strategy also permitted the incorporation of a well-characterised promoter upstream of each gene for expression in a heterologous host. The results demonstrate how site-specific recombination based on orthogonal serine integrases can be applied in Streptomyces spp. KeAi Publishing 2020-06-18 /pmc/articles/PMC7306541/ /pubmed/32596521 http://dx.doi.org/10.1016/j.synbio.2020.05.006 Text en © 2020 KeAi Communications Co.(+) Ltd http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Gao, Hong
Taylor, Gabrielle
Evans, Stephanie K.
Fogg, Paul C.M.
Smith, Margaret C.M.
Application of serine integrases for secondary metabolite pathway assembly in Streptomyces
title Application of serine integrases for secondary metabolite pathway assembly in Streptomyces
title_full Application of serine integrases for secondary metabolite pathway assembly in Streptomyces
title_fullStr Application of serine integrases for secondary metabolite pathway assembly in Streptomyces
title_full_unstemmed Application of serine integrases for secondary metabolite pathway assembly in Streptomyces
title_short Application of serine integrases for secondary metabolite pathway assembly in Streptomyces
title_sort application of serine integrases for secondary metabolite pathway assembly in streptomyces
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7306541/
https://www.ncbi.nlm.nih.gov/pubmed/32596521
http://dx.doi.org/10.1016/j.synbio.2020.05.006
work_keys_str_mv AT gaohong applicationofserineintegrasesforsecondarymetabolitepathwayassemblyinstreptomyces
AT taylorgabrielle applicationofserineintegrasesforsecondarymetabolitepathwayassemblyinstreptomyces
AT evansstephaniek applicationofserineintegrasesforsecondarymetabolitepathwayassemblyinstreptomyces
AT foggpaulcm applicationofserineintegrasesforsecondarymetabolitepathwayassemblyinstreptomyces
AT smithmargaretcm applicationofserineintegrasesforsecondarymetabolitepathwayassemblyinstreptomyces