Biosynthesis of Chlorinated Lactylates in Sphaerospermopsis sp. LEGE 00249

10.26434/chemrxiv.12885476 ◽

2020 ◽

Author(s):

Kathleen Abt ◽

Raquel Castelo-Branco ◽

Pedro Leao

Keyword(s):

Gene Cluster ◽

Building Blocks ◽

Bioinformatic Analysis ◽

Biosynthetic Gene Cluster ◽

Pathway Engineering ◽

Dodecanoic Acid ◽

Biosynthetic Gene ◽

Precursor Feeding ◽

Alkyl Chains ◽

Important Group

Lactylates are an important group of molecules in the food and cosmetic industries. A series of natural halogenated 1-lactylates – chlorosphaerolactyaltes (1-4) – were recently reported from Sphaerospermopsis sp. LEGE 00249. Here, we identify the cly biosynthetic gene cluster, containing all the necessary functionalities to generate and release the natural lactylates. Using a combination of stable isotope-labeled precursor feeding and bioinformatic analysis, we propose that dodecanoic acid and pyruvate are the key building blocks in the biosynthesis of 1-4. We additionally report minor analogues of these molecules with varying alkyl chains. The discovery of the cly gene cluster paves the way to accessing industrially-relevant lactylates through pathway engineering.

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Biosynthesis of Chlorinated Lactylates in Sphaerospermopsis Sp. LEGE 00249

10.26434/chemrxiv.12885476.v1 ◽

2020 ◽

Author(s):

Kathleen Abt ◽

Raquel Castelo-Branco ◽

Pedro Leao

Keyword(s):

Gene Cluster ◽

Building Blocks ◽

Bioinformatic Analysis ◽

Biosynthetic Gene Cluster ◽

Pathway Engineering ◽

Dodecanoic Acid ◽

Biosynthetic Gene ◽

Precursor Feeding ◽

Alkyl Chains ◽

Important Group

Lactylates are an important group of molecules in the food and cosmetic industries. A series of natural halogenated 1-lactylates – chlorosphaerolactyaltes (1-4) – were recently reported from Sphaerospermopsis sp. LEGE 00249. Here, we identify the cly biosynthetic gene cluster, containing all the necessary functionalities to generate and release the natural lactylates. Using a combination of stable isotope-labeled precursor feeding and bioinformatic analysis, we propose that dodecanoic acid and pyruvate are the key building blocks in the biosynthesis of 1-4. We additionally report minor analogues of these molecules with varying alkyl chains. The discovery of the cly gene cluster paves the way to accessing industrially-relevant lactylates through pathway engineering.

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Characterisation and heterologous biosynthesis of burnettiene A, a new polyene-decalin polyketide from Aspergillus burnettii

10.33774/chemrxiv-2021-p016t ◽

2021 ◽

Author(s):

Indra Roux ◽

Simon Bowles ◽

John A. Kalaitzis ◽

Daniel Vuong ◽

Ernest Lacey ◽

...

Keyword(s):

Gene Cluster ◽

Bioinformatic Analysis ◽

Biosynthetic Gene Cluster ◽

Tricarboxylic Acid ◽

In Vitro Cytotoxicity ◽

Methyl Groups ◽

Biosynthetic Gene ◽

Heterologous Biosynthesis ◽

Mouse Myeloma

Chemical exploration of the recently described Australian fungus, Aspergillus burnettii, uncovered a new metabolite, burnettiene A. Here, we characterise the structure of burnettiene A as a polyene-decalin polyketide. Bioinformatic analysis of the genome of A. burnettii identified a putative biosynthetic gene cluster for burnettiene A (bue), consisting of eight genes and sharing similarity to the fusarielin gene cluster. Introduction of the reassembled bue gene cluster into Aspergillus nidulans for heterologous expression resulted in the production of burnettiene A under native promoters. Omission of bueE encoding a cytochrome P450 led to the production of preburnettiene A, confirming that BueE is responsible for catalysing the regiospecific multi-oxidation of terminal methyl groups to carboxylic acids. Similarly, bueF was shown to encode an ester-forming methyltransferase, with its omission resulting in the production of the tricarboxylic acid, preburnettiene B. Introduction of an additional copy of the transcription factor bueR under the regulation of the gpdA promoter significantly improved the heterologous production of the burnettienes. Burnettiene A displayed strong in vitro cytotoxicity against mouse myeloma NS-1 cells (MIC 0.8 µg/mL).

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Organization of the biosynthetic gene cluster for the macrolide concanamycin A in Streptomyces neyagawaensis ATCC 27449

Microbiology ◽

10.1099/mic.0.28194-0 ◽

2005 ◽

Vol 151 (10) ◽

pp. 3161-3169 ◽

Cited By ~ 59

Author(s):

Stephen F. Haydock ◽

Anthony N. Appleyard ◽

Tatiana Mironenko ◽

John Lester ◽

Natasha Scott ◽

...

Keyword(s):

Gene Cluster ◽

Biosynthetic Pathway ◽

Polyketide Synthase ◽

Building Blocks ◽

Biosynthetic Gene Cluster ◽

Sequence Motif ◽

Biosynthetic Gene ◽

Concanamycin A ◽

Modular Polyketide Synthase

The macrolide antibiotic concanamycin A has been identified as an exceptionally potent inhibitor of the vacuolar (V-type) ATPase. Such compounds have been mooted as the basis of a potential drug treatment for osteoporosis, since the V-ATPase is involved in the osteoclast-mediated bone resorption that underlies this common condition. To enable combinatorial engineering of altered concanamycins, the biosynthetic gene cluster governing the biosynthesis of concanamycin A has been cloned from Streptomyces neyagawaensis and shown to span a region of over 100 kbp of contiguous DNA. An efficient transformation system has been developed for S. neyagawaensis and used to demonstrate the role of the cloned locus in the formation of concanamycin A. Sequence analysis of the 28 ORFs in the region has revealed key features of the biosynthetic pathway, in particular the biosynthetic origin of portions of the backbone, which arise from the unusual polyketide building blocks ethylmalonyl-CoA and methoxymalonyl-ACP, and the origin of the pendant deoxysugar moiety 4′-O-carbamoyl-2′-deoxyrhamnose, as well as the presence of a modular polyketide synthase (PKS) encoded by six giant ORFs. Examination of the methoxymalonyl-specific acyltransferase (AT) domains has led to recognition of an amino acid sequence motif which can be used to distinguish methylmalonyl-CoA- from methoxymalonyl-ACP-specific AT domains in natural PKSs.

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Identifying the Biosynthetic Gene Cluster for Triacsins with an N-hydroxytriazene Moiety

10.1101/495424 ◽

2018 ◽

Author(s):

Frederick F. Twigg ◽

Wenlong Cai ◽

Wei Huang ◽

Joyce Liu ◽

Michio Sato ◽

...

Keyword(s):

Biological Activity ◽

Lipid Metabolism ◽

Gene Cluster ◽

Gene Disruption ◽

Bioinformatic Analysis ◽

Biosynthetic Gene Cluster ◽

Biosynthetic Gene ◽

Streptomyces Tsukubaensis ◽

Targeted Gene Disruption

AbstractTriacsins are a family of natural products containing an N-hydroxytriazene moiety not found in any other known secondary metabolites. Though many studies have examined the biological activity of triacsins in lipid metabolism, the biosynthesis of triacsins has remained unknown. Here, we report the identification of the triacsin biosynthetic gene cluster in Streptomyces aureofaciens ATCC 31442. Bioinformatic analysis of the gene cluster led to the discovery of the tacrolimus producer Streptomyces tsukubaensis NRRL 18488 as a new triacsin producer. In addition to targeted gene disruption to identify necessary genes for triacsin production, stable isotope feeding was performed in vivo to advance the understanding of N-hydroxytriazene biosynthesis.

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New Insights into Chloramphenicol Biosynthesis in Streptomyces venezuelae ATCC 10712

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.04272-14 ◽

2014 ◽

Vol 58 (12) ◽

pp. 7441-7450 ◽

Cited By ~ 45

Author(s):

Lorena T. Fernández-Martínez ◽

Chiara Borsetto ◽

Juan Pablo Gomez-Escribano ◽

Maureen J. Bibb ◽

Mahmoud M. Al-Bassam ◽

...

Keyword(s):

Gene Cluster ◽

Acyl Carrier Protein ◽

Bioinformatic Analysis ◽

Biosynthetic Gene Cluster ◽

Transcriptional Analysis ◽

Biosynthetic Gene ◽

Phosphopantetheinyl Transferase ◽

Streptomyces Venezuelae ◽

Content Type ◽

New Genes

ABSTRACTComparative genome analysis revealed seven uncharacterized genes,sven0909tosven0915, adjacent to the previously identified chloramphenicol biosynthetic gene cluster (sven0916–sven0928) ofStreptomyces venezuelaestrain ATCC 10712 that was absent in a closely relatedStreptomycesstrain that does not produce chloramphenicol. Transcriptional analysis suggested that three of these genes might be involved in chloramphenicol production, a prediction confirmed by the construction of deletion mutants. These three genes encode a cluster-associated transcriptional activator (Sven0913), a phosphopantetheinyl transferase (Sven0914), and a Na+/H+antiporter (Sven0915). Bioinformatic analysis also revealed the presence of a previously undetected gene,sven0925, embedded within the chloramphenicol biosynthetic gene cluster that appears to encode an acyl carrier protein, bringing the number of new genes likely to be involved in chloramphenicol production to four. Microarray experiments and synteny comparisons also suggest thatsven0929is part of the biosynthetic gene cluster. This has allowed us to propose an updated and revised version of the chloramphenicol biosynthetic pathway.

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Anacyclamide D8P, a prenylated cyanobactin from a Sphaerospermopsis sp. cyanobacterium

10.26434/chemrxiv.5346739.v1 ◽

2017 ◽

Cited By ~ 1

Author(s):

Joana Martins ◽

Niina Leikoski ◽

Matti Wahlsten ◽

Joana Azevedo ◽

Jorge Antunes ◽

...

Keyword(s):

Gene Cluster ◽

Cyclic Peptides ◽

Biosynthetic Gene Cluster ◽

The Novel ◽

Tyrosine Residue ◽

Biosynthetic Gene ◽

Post Translational Modification ◽

Biological Assays ◽

Mass Spectrometric Data ◽

Spectrometric Data

Cyanobactins are a family of linear and cyclic peptides produced through the post-translational modification of short precursor peptides. Anacyclamides are macrocyclic cyanobactins with a highly diverse sequence that are common in the genus Anabaena. A mass spectrometry-based screening of potential cyanobactin producers led to the discovery of a new prenylated member of this family of compounds, anacyclamide D8P (1), from Sphaerospermopsis sp. LEGE 00249. The anacyclamide biosynthetic gene cluster (acy) encoding the novel macrocyclic prenylated cyanobactin, was sequenced. Heterologous expression of the acy gene cluster in Escherichia coli established the connection between genomic and mass spectrometric data. Unambiguous establishment of the type and site of prenylation required the full structural elucidation of 1 using Nuclear Magnetic Resonance (NMR), which demonstrated that a forward prenylation occurred on the tyrosine residue. Compound 1 was tested in pharmacologically or ecologically relevant biological assays and revealed moderate antimicrobial activity towards the fouling bacterium Halomonas aquamarina CECT 5000.

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