High production of a class III lantipeptide AmfS in Streptomyces griseus

Abstract

AmfS, a class III lantipeptide serves as a morphogen in Streptomyces griseus. Here, we constructed a high production system of AmfS in S. griseus. We isolated S. griseus Grd1 strain defective in glucose repression of aerial mycelium formation and found it suitable for the overproduction of AmfS. Two expression vectors carrying the strong and constitutive ermE2 promoter were constructed using a multicopy number plasmid, pIJ702. The use of the Grd1 strain combined with the expression vectors enabled high production of AmfS by S. griseus into its culture broth. The expression system was also effective for the generation of abundant AmfS derived from Streptomyces avermitilis. In addition, site-directed mutagenesis revealed the amino acid residues essential for the morphogen activity of AmfS. These results indicate that the constructed system enables efficient production of class III lantipeptides by Streptomyces.

Mechanism of Action of Ribosomally Synthesized and Post-Translationally Modified Peptides

“…Several other peptides with sequence homology to SapB have been identified including AmfS ( 36 ), produced in Streptomyces griseus (Figure ). , AmfS is also important for aerial hyphae formation and sporulation. , Mutagenesis studies of AmfS revealed that all conserved post-translationally modified residues selected for replacement (Ser3, Cys10, Ser13, Ser16, and Cys20) were essential for activity . Ser6, although conserved and converted to Dha, was not targeted by mutagenesis in this study.…”

A Multidisciplinary Approach to Unraveling the Natural Product Biosynthetic Potential of a Streptomyces Strain Collection Isolated from Leaf-Cutting Ants

“…The remaining 21 compounds that were not detected by dereplication were mainly classified as siderophores, terpenes, lanthipeptides, lasso peptides, lipopeptides, ectoine, butyrolactone and melanin. Our inability to detect these compounds could be due to two different reasons: (i) they were not being produced under our laboratory conditions or (ii) the extraction method using ethyl acetate is not suitable for this type of metabolites, and other solvents (e.g., n-butanol or ethanol) should be used instead [49][50][51][52].…”

De novo biosynthesis of garbanzol and fustin in Streptomyces albus based on a potential flavanone 3‐hydroxylase with 2‐hydroxylase side activity

“…2A). The ermE * promoter is one of the most widely used promoters for heterologous gene expression in streptomycetes (Lombó et al ., 2006; Park et al ., 2009; Wang et al ., 2012; Takano et al ., 2017; Liu et al ., 2018), and its correct activity for controlling flavonoid biosynthetic genes has been confirmed in previous studies of this research group (Marín et al ., 2017, 2018; García‐Gutiérrez et al ., 2020). Genes were selected from different bacteria ( Rhodobacter capsulatus and S. coelicolor ) and plants ( Glycine max, Petroselinum crispum and Arabidopsis thaliana ) as they were already tested in the laboratory for these enzymatic steps (Marín et al ., 2017, 2018; García‐Gutiérrez et al ., 2020).…”