Heterologous expression of the diazaquinomycin biosynthetic gene cluster

Abstract

Members of the diazaquinomycin class of natural products have shown potent and selective activity against Mycobacterium tuberculosis. However, poor aqueous solubility has prevented extensive studies in animal models thus far. Our long-term goal is to harness knowledge regarding diazaquinomycin biosynthesis towards the generation of derivatives for structure–activity relationship studies. We have previously sequenced the genomes of two diazaquinomycin-producing, actinomycete bacteria and identified putative daq biosynthetic gene clusters. Here, we report the heterologous expression of the daq gene cluster from the marine Streptomyces sp. F001 in S. coelicolor M1152. In addition to serving as functional proof for gene cluster assignment, the heterologous expression system reported here is expected to facilitate investigations aimed at elucidating diazaquinomycin biosynthesis.

Enzyme-Constrained Models and Omics Analysis of Streptomyces coelicolor Reveal Metabolic Changes that Enhance Heterologous Production

“…This mutation was shown to have strong positive effects on the production of various antibiotics ( Gomez-Escribano and Bibb, 2011 ; Hu et al., 2002 ). Up to now, M1152 is a preferred general “superhost” for heterologous BGC expression ( Braesel et al., 2019 ; Castro et al., 2015 ; Kepplinger et al., 2018 ; Li et al., 2013 ; Thanapipatsiri et al., 2015 ) and is the starting point for further strain development.…”

Deciphering Deoxynybomycin Biosynthesis Reveals Fe(II)/α-Ketoglutarate-Dependent Dioxygenase-Catalyzed Oxazoline Ring Formation and Decomposition

“…The proteins encoded in the dnm BGC show high homology to those in the nyb BGC 17 with the amino acid identity of 60−86% (except the 48% identity of DnmM and NybM) (Table S2), and the proteins encoded in the dnq BGC also exhibit high similarity to those in daq BGCs from Streptomyces sp. F001 32,33 (Figure 2A) with identity in amino acid sequences of 38−67% (Table S3). The highly conserved architectures of dnm, nyb, dnq, and daq BGCs indicated that the biosynthesis of DNM (1) and DNQ (3) should share a common enzymatic system to construct the tricyclic ring system, as previously proposed for NM (2) and DAQs (Figure 2B).…”

Advances in Biosynthesis of Natural Products from Marine Microorganisms

“…Although the yield was similar to that of the original host, the culturing efficiency was improved significantly to allow a simpler scale-up of the experiment. Moreover, the heterologous biosynthetic system provides the basis for generating structural analogs to solve the low solubility problem [78].…”

“…Although the yield was similar to that of the original host, the culturing efficiency was improved significantly to allow a simpler scale-up of the experiment. Moreover, the heterologous biosynthetic system provides the basis for generating structural analogs to solve the low solubility problem [78]. Escherichia coli is frequently used as a heterologous host of choice due to its fast growth, abundant genetic tools, and the deeper understanding we have of its metabolism.…”