Identification of the Anti-Infective Aborycin Biosynthetic Gene Cluster from Deep-Sea-Derived Streptomyces sp. SCSIO ZS0098 Enables Production in a Heterologous Host

Abstract

Aborycin is a ribosomally synthesized member of the type I lasso peptide natural products. In the present study, aborycin was isolated and identified from the deep-sea-derived microbe Streptomyces sp. SCSIO ZS0098. The aborycin biosynthetic gene cluster (abo) was identified on the basis of genome sequence analyses and then heterologously expressed in Streptomyces coelicolor M1152 to effectively produce aborycin. Aborycin generated in this fashion exhibited moderate antibacterial activity against 13 Staphylococcus aureus strains from various sources with minimum inhibitory concentrations MICs = 8.0~128 µg/mL, against Enterococcus faecalis ATCC 29212 with an MIC = 8.0 µg/mL, and against Bacillus thuringiensis with MIC = 2.0 µg/mL. Additionally, aborycin displayed potent antibacterial activity (MIC = 0.5 µg/mL) against the poultry pathogen Enterococcus gallinarum 5F52C. The reported abo cluster clearly has the potential to provide a means of expanding the repertoire of anti-infective type I lasso peptides.

Heterologous Production of Lasso Peptide Capistruin in a Burkholderia Host

“…According to the reported codon preferences for Burkholderia mallei, 25 the synonymous modification introduced is expected to be well tolerated. Transcription was driven from the L-arabinose inducible araC/PBAD promoter that had been previously shown to work well in FERM BP-3421, 11 which does not utilize L-arabinose as a carbon source. 9 The placement of the transcriptional start site of PBAD was 45 bp from the RBS, which was comparable to the location of the native promoter.…”

“…9 The placement of the transcriptional start site of PBAD was 45 bp from the RBS, which was comparable to the location of the native promoter. The araC/PBAD fragment was first ligated into the E. coli-Burkholderia replicative vector pUCP_neo 11 to yield pSK019, after which synthetic capABCD was inserted into pSK019 to yield pSK020 (Figure 1 and Figure S1). Figure 1.…”

“…The native promoter was replaced with the araC/PBAD promoter that had been shown to work in FERM BP-3421. 11 The sequence of the promoter is annotated in red as reported by Greenfield et al 26 The distance between the transcriptional start site Establishment of an electroporation protocol. The only reported DNA transfer method for FERM BP-3421 is conjugation from E. coli.…”

“…9,10 Strain FERM BP-3421 was previously developed to produce native polyketide-nonribosomal peptides of the spliceostatin class of natural products in industrial scale and in up to 6 g/L yields. 11 As a model BGC to test the host with, we chose the BGC encoding biosynthesis of the lasso peptide capistruin. Our main rationale for choosing the capistruin BGC was because it had been previously expressed in E. coli, allowing us to determine whether our chosen host offers any advantage over the broadly used E. coli host.…”

Identification and Heterologous Expression of the Biosynthetic Gene Cluster Encoding the Lasso Peptide Humidimycin, a Caspofungin Activity Potentiator

“…The BGCs of additional class I siamycin-like molecules (aborycin, specialicin, and MS-271) have been recently identified by genome mining, and some of them have been heterologously expressed [28][29][30].…”