Re-engineering robustness in to industrial antibiotic producing Streptomyces strains

Lead Research Organisation: University of Strathclyde
Department Name: Inst of Pharmacy and Biomedical Sci

Abstract

Currently around two-thirds of our antibiotics are made by the soil bacterium Streptomyces. Industrial Streptomyces usually undergo several rounds of random mutagenesis to improve their production characteristics becoming highly-adapted to industrial culture conditions. There is a problem, however, highly-adapted strains are less flexible limiting alterations to production processes and the introduction of sustainable feedstocks. This project will identify genes used by the bacteria to adapt to certain feedstocks and develop new, rapid methods to repair genes lost or damaged genes during mutagenesis, maintaining performance and increasing the flexibility of high-producing strains. This is important because the generation of industrial strains is time consuming and labour intensive. Understanding what limits flexibility in high-yielding antibiotic producers and repairing these mutations will make it easier and greener to make future antibiotics and combat antimicrobial resistance.
 
Title Supplementary data for McHugh et al., 2022 - Biosynthesis of aurodox, a Type III secretion system inhibitor from Streptomyces goldiniensis. 
Description The global increase in antimicrobial-resistant infections means that there is a need to develop new antimicrobial molecules and strategies to combat the issue. Aurodox is a linear polyketide natural product that is produced by Streptomyces goldiniensis, yet little is known about aurodox biosynthesis or the nature of the biosynthetic gene cluster (BGC) that encodes its production. To gain a deeper understanding of aurodox biosynthesis by S. goldiniensis, the whole genome of the organism was sequenced, revealing the presence of an 87 kb hybrid Polyketide Synthase/Non-Ribosomal Peptide Synthetase (PKS/NRPS) BGC. The aurodox BGC shares significant homology with the kirromycin BGC from S. collinus T? 365; however, the genetic organisation of the BGC differs significantly. The candidate aurodox gene cluster was cloned and expressed in a heterologous host to demonstrate that it was responsible for aurodox biosynthesis and disruption of the primary PKS gene (aurAI) abolished aurodox production. These data support a model whereby the initial core biosynthetic reactions involved in aurodox biosynthesis follow that of kirromycin. Cloning aurM* from S. goldiniensis and expressing this in the kirromycin producer S. collinus T? 365 enabled methylation of the pyridone group, suggesting this is the last step in biosynthesis. This methylation step is also sufficient to confer the unique Type III Secretion System inhibitory properties to aurodox. 
Type Of Art Image 
Year Produced 2022 
URL https://figshare.com/articles/figure/Supplementary_data_for_McHugh_et_al_2022_-_Biosynthesis_of_auro...
 
Description We have developed in collaboration with The Game Doctor Limited a game that allows 11-18 year old students and interested parties to simulate the antibiotic discovery process from isolation of bacteria through to chemical analysis - the app is soft launched (march 2nd 2021) but we will be using a stronger social media presence during science week (5-14th March 2021) in the UK to promote the game - it is available as an iPhone/iPad app App: https://apps.apple.com/gb/app/dr-dirt/id1543940322 WebGL version: https://www.drdirt.co.uk
First Year Of Impact 2021
Sector Education
Impact Types Cultural
 
Title Genome sequence of the aurodox-producing bacterium Streptomyces goldiniensis ATCC 21386 - ACMI-D-22-00033R1 
Description Genome sequence of the aurodox-producing bacterium Streptomyces goldiniensis ATCC 21386 Access Microbiology manuscript ACMI-D-22-00033R1We report the genome sequence of Streptomyces goldiniensis ATCC 21386, a strain which produces the anti-bacterial and anti-virulence polyketide, aurodox. The genome of S. goldiniensis ATCC 21386 was sequenced using a multiplatform hybrid approach, revealing a linear genome of ~10 Mbp with a G+C content of 71 %. The genome sequence revealed 36 putative biosynthetic gene clusters (BGCs), including a large region of 271 Kbp that was rich in biosynthetic capability. The genome sequence is deposited in DDBJ/EMBL/GenBank with the accession number PRJNA602141.These data represent the FastQ outputs from the Oxford Nanopore sequencing that contributed to the hybrid genome assembly. 
Type Of Material Database/Collection of data 
Year Produced 2022 
Provided To Others? Yes  
URL https://figshare.com/articles/dataset/Genome_sequence_of_the_aurodox-producing_bacterium_Streptomyce...