GPAHex-A synthetic biology platform for Type IV–V glycopeptide antibiotic production and discovery

Abstract

Glycopeptide antibiotics (GPAs) are essential for the treatment of severe infectious diseases caused by Gram-positive bacteria. The emergence and spread of GPA resistance have propelled the search for more effective GPAs. Given their structural complexity, genetic intractability, and low titer, expansion of GPA chemical diversity using synthetic or medicinal chemistry remains challenging. Here we describe a synthetic biology platform, GPAHex (GPA Heterologous expression), which exploits the genes required for the specialized GPA building blocks, regulation, antibiotic transport, and resistance for the heterologous production of GPAs. Application of the GPAHex platform results in: (1) a 19-fold increase of corbomycin titer compared to the parental strain, (2) the discovery of a teicoplanin-class GPA from an Amycolatopsis isolate, and (3) the overproduction and characterization of a cryptic nonapeptide GPA. GPAHex provides a platform for GPA production and mining of uncharacterized GPAs and provides a blueprint for chassis design for other natural product classes.

Synthetic Biology Tools for Engineering Microbial Cells to Fight Superbugs

“…has poor performance ( Culp et al, 2020 ). Xu et al used a glycopeptide antibiotic heterologous expression system (GPAHex) to enhance the expression of genes for corbomicyn synthesis in S. coelicolor , obtaining a 19-fold increase in titers using this platform ( Xu et al, 2020 ).…”

Genomic Insights into the Distribution and Phylogeny of Glycopeptide Resistance Determinants within the Actinobacteria Phylum

“…Type V GPAs are not glycopeptides sensu stricto , since they are not glycosylated; moreover, they are not dalbaheptides, since they include also nonapeptide antibiotics (such as corbomycin and GP6738 [ 19 , 20 ]), and they target autolysins (named also murein hydrolases) instead of lipid II. Autolysins are enzymes breaking the bonds within the peptidoglycan to allow bacterial growth and cell division [ 21 ]; thus, type V GPAs block cell-wall remodeling, arresting cell division.…”

Chemotranscriptomic Profiling Defines Drug-Specific Signatures of the Glycopeptide Antibiotics Dalbavancin, Vancomycin and Chlorobiphenyl-Vancomycin in a VanB-Type-Resistant Streptomycete

“…VanB-type resistance is defined by inducible resistance to vancomycin but susceptibility to teicoplanin and related analogues (unless vancomycin is also present). Routes to the production of new glycopeptide antibiotics with improved efficacy are continually being sought and include total chemical synthesis ( Okano et al, 2014 , 2017 ), chemical or chemoenzymatic derivatisation of glycopeptide scaffolds ( Yarlagadda et al, 2015 ; Blaskovich et al, 2018 ; MishraI, Stolarzewicz et al, 2018 ; Tailhades et al, 2020 ), and the development of synthetic biology platforms ( Xu et al, 2020 ). Successful modification of a glycopeptide structure might result from the generation of molecules with reduced ability to activate the known inducible glycopeptide resistance systems or the conferment of antibacterial activities independent of the interaction with d-Ala–d-Ala residues.…”

“…VanB-type resistance is defined by inducible resistance to vancomycin but susceptibility to teicoplanin and related analogues (unless vancomycin is also present). Routes to the production of new glycopeptide antibiotics with improved efficacy are continually being sought and include total chemical synthesis (Okano et al, 2014(Okano et al, , 2017, chemical or chemoenzymatic derivatisation of glycopeptide scaffolds (Yarlagadda et al, 2015;Blaskovich et al, 2018;MishraI, Stolarzewicz et al, 2018;Tailhades et al, 2020), and the development of synthetic biology platforms (Xu et al, 2020).…”