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 orthogonal...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.展开更多
基金This work was supported by the Biotechnology and Biological Sciences Research Council project grant BB/K003356/1.HPLC-MS analysis was performed on equipment within the Centre of Excellence in Mass Spectrometry(University of York),funded by Science City York(Yorkshire Forward,EP/K039660/1,EP/M028127/1).
文摘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.
