Biosynthesis of Chlorinated Lactylates in Sphaerospermopsis sp. LEGE 00249

Abt, Kathleen; Castelo-Branco, Raquel; Leão, Pedro N.

doi:10.26434/chemrxiv.12885476

Cited by 3 publications

(4 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In our previous study, 6 nocuolactylates were found to incorporate three alkyl moieties derived from either hexanoic acid or longer fatty acids. The lactylate portion of these molecules accounts for one such incorporation, 12 indicating that the two remaining moieties are used for the biosynthesis of 1. We considered that these building blocks could be used to generate the C13 alkyl moiety of 1.…”

Section: Annotation Of the Noc Gene Products And Acyl-acp Dehydrogena...mentioning

confidence: 99%

“…The latter has been implicated in the biosynthesis of the chlorosphaerolactylates and was predicted to activate dodecanoic acid, but also decanoic and tetradecanoic acids to a minor extent. 12 We heterologously expressed and purified each FAAL as well as with their cognate ACPs in E. coli. NocH contains both FAAL and ACP domains and was expressed as a single polypeptide, while ClyA and its associated ACP ClyB were expressed and purified separately.…”

Section: Noch Activates Hexanoic and Octanoic Acidsmentioning

confidence: 99%

“…1b). 4,12 However, several of the putative noc genes have recently been implicated in the biosynthesis of chlorosphaerolactylates (e.g. 2, Fig.…”

Section: Introductionmentioning

confidence: 99%

“…1b). 12 To acknowledge these findings, those genes were renamed as clyA-F, constituting the cly BGC. Our group later showed that Nodularia sp.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Biosynthesis of the unusual carbon skeleton of nocuolin A

Martins

Glasser

Kountz

et al. 2022

Preprint

View full text Add to dashboard Cite

Nocuolin A is a cytotoxic cyanobacterial metabolite that is proposed to be produced by enzymes of the noc biosynthetic gene cluster. Nocuolin A features a 1,2,3-oxadiazine moiety, a structural feature unique among natural products and, so far, inaccessible through organic synthesis, suggesting that novel enzymatic chemistry might be involved in its biosynthesis. This heterocycle is substituted with two alkyl chains and a 3-hydroxypropanoyl moiety. We report here our efforts to elucidate the origin of the carbon skeleton of nocuolin A. Supplementation of cyanobacterial cultures with stable isotope-labeled fatty acids revealed that the central C13 chain is assembled from two medium-chain fatty acids, hexanoic and octanoic acids. Using biochemical assays, we show that a fatty acyl-AMP ligase, NocH, activates both fatty-acids as acyl adenylates, which are loaded onto acyl carrier protein domains and undergo a non-decarboxylative Claisen condensation catalyzed by the ketosynthase NocG. This enzyme is part of a phylogenetically well-defined clade within similar genomic contexts. NocG presents a unique combination of characteristics found in other ketosynthases, namely in terms of substrate specificity and reactivity. Further supplementation experiments indicate that the 3-hydroxypropanoyl moiety of 1 originates from methionine, through an as-yet-uncharacterized mechanism. This work provides ample biochemical evidence connecting the putative noc biosynthetic gene cluster to nocuolin A and identifies the origin of all its carbon atoms, setting the stage for elucidation of its unusual biosynthetic chemistry.

show abstract

Section: Annotation Of the Noc Gene Products And Acyl-acp Dehydrogena...mentioning

confidence: 99%

Section: Noch Activates Hexanoic and Octanoic Acidsmentioning

confidence: 99%

“…1b). 4,12 However, several of the putative noc genes have recently been implicated in the biosynthesis of chlorosphaerolactylates (e.g. 2, Fig.…”

Section: Introductionmentioning

confidence: 99%

“…1b). 12 To acknowledge these findings, those genes were renamed as clyA-F, constituting the cly BGC. Our group later showed that Nodularia sp.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Biosynthesis of the unusual carbon skeleton of nocuolin A

Martins

Glasser

Kountz

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

Distribution and diversity of dimetal-carboxylate halogenases in cyanobacteria

Eusébio

Rego

Glasser

et al. 2021

Preprint

View full text Add to dashboard Cite

Halogenation is a recurring feature in natural products, especially those from marine organisms. The selectivity with which halogenating enzymes act on their substrates renders halogenases interesting targets for biocatalyst development. Recently, CylC – the first predicted dimetal-carboxylate halogenase to be characterized – was shown to regio- and stereoselectively install a chlorine atom onto an unactivated carbon center during cylindrocyclophane biosynthesis. Homologs of CylC are also found in other characterized cyanobacterial secondary metabolite biosynthetic gene clusters. Due to its novelty in biological catalysis, selectivity and ability to perform C-H activation, this halogenase class is of considerable fundamental and applied interest. However, little is known regarding the diversity and distribution of these enzymes in bacteria. In this study, we used both genome mining and PCR-based screening to explore the genetic diversity and distribution of CylC homologs. While we found non-cyanobacterial homologs of these enzymes to be rare, we identified a large number of genes encoding CylC-like enzymes in publicly available cyanobacterial genomes and in our in-house culture collection of cyanobacteria. Genes encoding CylC homologs are widely distributed throughout the cyanobacterial tree of life, within biosynthetic gene clusters of distinct architectures. Their genomic contexts feature a variety of biosynthetic partners, including fatty-acid activation enzymes, type I or type III polyketide synthases, dialkylresorcinol-generating enzymes, monooxygenases or Rieske proteins. Our study also reveals that dimetal-carboxylate halogenases are among the most abundant types of halogenating enzymes in the phylum Cyanobacteria. This work will help to guide the search for new halogenating biocatalysts and natural product scaffolds.Data statementAll supporting data and methods have been provided within the article or through a Supplementary Material file, which includes 14 supplementary figures and 4 supplementary tables.

show abstract

Distribution and diversity of dimetal-carboxylate halogenases in cyanobacteria

et al. 2021

Self Cite

View full text Add to dashboard Cite

Background Halogenation is a recurring feature in natural products, especially those from marine organisms. The selectivity with which halogenating enzymes act on their substrates renders halogenases interesting targets for biocatalyst development. Recently, CylC – the first predicted dimetal-carboxylate halogenase to be characterized – was shown to regio- and stereoselectively install a chlorine atom onto an unactivated carbon center during cylindrocyclophane biosynthesis. Homologs of CylC are also found in other characterized cyanobacterial secondary metabolite biosynthetic gene clusters. Due to its novelty in biological catalysis, selectivity and ability to perform C-H activation, this halogenase class is of considerable fundamental and applied interest. The study of CylC-like enzymes will provide insights into substrate scope, mechanism and catalytic partners, and will also enable engineering these biocatalysts for similar or additional C-H activating functions. Still, little is known regarding the diversity and distribution of these enzymes. Results In this study, we used both genome mining and PCR-based screening to explore the genetic diversity of CylC homologs and their distribution in bacteria. While we found non-cyanobacterial homologs of these enzymes to be rare, we identified a large number of genes encoding CylC-like enzymes in publicly available cyanobacterial genomes and in our in-house culture collection of cyanobacteria. Genes encoding CylC homologs are widely distributed throughout the cyanobacterial tree of life, within biosynthetic gene clusters of distinct architectures (combination of unique gene groups). These enzymes are found in a variety of biosynthetic contexts, which include fatty-acid activating enzymes, type I or type III polyketide synthases, dialkylresorcinol-generating enzymes, monooxygenases or Rieske proteins. Our study also reveals that dimetal-carboxylate halogenases are among the most abundant types of halogenating enzymes in the phylum Cyanobacteria. Conclusions Our data show that dimetal-carboxylate halogenases are widely distributed throughout the Cyanobacteria phylum and that BGCs encoding CylC homologs are diverse and mostly uncharacterized. This work will help guide the search for new halogenating biocatalysts and natural product scaffolds.

show abstract

Biosynthesis of Chlorinated Lactylates in Sphaerospermopsis sp. LEGE 00249

Cited by 3 publications

References 31 publications

Biosynthesis of the unusual carbon skeleton of nocuolin A

Biosynthesis of the unusual carbon skeleton of nocuolin A

Distribution and diversity of dimetal-carboxylate halogenases in cyanobacteria

Distribution and diversity of dimetal-carboxylate halogenases in cyanobacteria

Contact Info

Product

Resources

About