Biosynthesis of Colabomycin E, a New Manumycin‐Family Metabolite, Involves an Unusual Chain‐Length Factor

Abstract

Colabomycin E is a new member of the manumycin-type metabolites produced by the strain Streptomyces aureus SOK1/5-04 and identified by genetic screening from a library of streptomycete strains. The structures of colabomycin E and accompanying congeners were resolved. The entire biosynthetic gene cluster was cloned and expressed in Streptomyces lividans. Bioinformatic analysis and mutagenic studies identified components of the biosynthetic pathway that are involved in the formation of both polyketide chains. Recombinant polyketide synthases (PKSs) assembled from the components of colabomycin E and asukamycin biosynthetic routes catalyzing the biosynthesis of "lower" carbon chains were constructed and expressed in S. aureus SOK1/5-04 ΔcolC11-14 deletion mutant. Analysis of the metabolites produced by recombinant strains provided evidence that in both biosynthetic pathways the length of the lower carbon chain is controlled by an unusual chain-length factor supporting biosynthesis either of a triketide in asukamycin or of a tetraketide in colabomycin E. Biological activity assays indicated that colabomycin E significantly inhibited IL-1β release from THP-1 cells and might thus potentially act as an anti-inflammatory agent.

Evolution of cyclizing 5-aminolevulinate synthases in the biosynthesis of actinomycete secondary metabolites: outcomes for genetic screening techniques

“…Sixty-six natural isolates were clustered in the linear polyketides branch, together with six uncharacterized genome-based sequences. Thirty eight of the natural isolates formed a separate branch represented by a novel colabomycin E producer, S. aureus SOK1/5-04 (BCCO10_5), which was identified by the genetic screening method described in this work and recently studied in detail by our group ( Petříčková et al, 2014 ). The next two isolates occurred in the manumycin-producer branch (BCCO10_215 and BCCO10_1636; isolated from Sokolov colliery spoil heaps and Mt.…”

“…Bacterial cultures were maintained long-term as spore suspensions in glycerol (15% v/v) at -80°C and/or freeze-dried. One group of strains had been isolated and characterized previously ( Chroňáková et al, 2010 ), consequently we studied one of these in greater detail ( S. aureus SOK 1/5-04, BCCO10_5, a new colabomycin E producer; Petříčková et al, 2014 ).…”

“…Both final extracts were combined, dissolved in acetone, and analyzed. UHPLC-DAD-ToF-MS analyses were carried out with a Waters Acquity UPLC System (Waters) using the Acquity UPLC BEH C18 column under the conditions specified in Petříčková et al (2014) .…”

A Human Lung-Associated Streptomyces sp. TR1341 Produces Various Secondary Metabolites Responsible for Virulence, Cytotoxicity and Modulation of Immune Response

“…Many strains produce cytotoxic or cytolytic compounds (Knopik-Skrocka and Bielawski, 2002), or compounds able to slow down or stop the host immune response, cell division, etc. (e.g., (Striz et al, 2008;Petrickova et al, 2014). People in regular contact with the soil dust (miners, farmers) as well as inhabitants of cities with high air pollution and water-damaged houses are the most exposed.…”

Production of a Novel Amide‐Containing Polyene by Activating a Cryptic Biosynthetic Gene Cluster in Streptomyces sp. MSC090213JE08

“…ORF3/SmcKSII and ORF2/SmcKSI belong to the same clades as Iga11 and Iga12, respectively . AsuC13/ColC13 (KS α ) and AsuC14/ColC14 (CLF) were reported to be unusual KSs involved in the biosynthesis of the polyene portion of the manumycin‐type compounds, asukamycin and colabomycin E . These KS α s and CLFs belong to phylogenetic clades different from those of Iga11 and Iga12.…”