BrtB is an O-alkylating enzyme that generates fatty acid-bartoloside esters

Reis, João P. A.; Figueiredo, Sandra A. C.; Sousa, Maria Lígia; Leão, Pedro N.

doi:10.1038/s41467-020-15302-z

Cited by 16 publications

(17 citation statements)

References 32 publications

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“… 22 These loci were associated with the biosynthesis of nocuolin A ( 6 , Figure 1 a) by Voráčová and co-workers, 22 based on comparative genomics (strains that contained the locus were found to produce 6 ). Despite the fact that CylC homologues are known to carry out cryptic halogenations and generate nonhalogenated products, 23 , 28 we considered that these two dimetal-carboxylate halogenases found in the LEGE 00249 genome were strong candidate enzymes for carrying out the halogenations in 1 – 4 .…”

Section: Resultsmentioning

confidence: 99%

Biosynthesis of Chlorinated Lactylates in Sphaerospermopsis sp. LEGE 00249

2021

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Lactylates are an important group of molecules in the food and cosmetic industries. A series of natural halogenated 1-lactylates, chlorosphaerolactylates ( 1 – 4 ), were recently reported from Sphaerospermopsis sp. LEGE 00249. Here, we identify the cly biosynthetic gene cluster, containing all the necessary functionalities for the biosynthesis of the natural lactylates, based on in silico analyses. Using a combination of stable isotope incorporation experiments 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. This work paves the way to accessing industrially relevant lactylates through pathway engineering.

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Section: Resultsmentioning

confidence: 99%

Biosynthesis of Chlorinated Lactylates in Sphaerospermopsis sp. LEGE 00249

2021

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“…These observations suggest that functionally convergent associations between CylC homologs and other proteins have emerged multiple times during evolution. Examples include the CylC/CylK and BrtJ/BrtB associations, which use cryptic halogenation to achieve C-C and C-O bond formation, respectively [27, 59]. However, the role of the CylC homolog-mediated halogenation of fatty acyl moieties observed for other cyanobacterial metabolites is not currently understood.…”

Section: Resultsmentioning

confidence: 99%

“…However, the role of the CylC homolog-mediated halogenation of fatty acyl moieties observed for other cyanobacterial metabolites is not currently understood. Interestingly, while a number of CylC homologs, including those that are part of characterized BGCs, likely act on ACP-tethered fatty acyl substrates [27, 59], those from the PriA-Rieske- and cytochrome P450/sulfotransferase categories do not have a neighboring carrier protein and therefore might not require a tethered substrate. This would be an important property for a CylC-like biocatalyst [15].…”

Section: Resultsmentioning

confidence: 99%

Distribution and diversity of dimetal-carboxylate halogenases in cyanobacteria

Eusébio

Rego

Glasser

et al. 2021

Preprint

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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.

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“…25 A recently reported cyanobacterial enzyme -BrtB -can also esterify free FAs with alkyl halide bearing NPs. 26 FAincorporating enzymes are abundant in cyanobacterial genomes (Fig. S1), often within orphan BGCs 22 and therefore represent opportunities for NP discovery.…”

Section: Introductionmentioning

confidence: 99%

“…27 On the other hand, different biosynthetic logics, tailoring enzymes and hypothetical proteins are found associated with these FA-incorporating enzymes. 22,26 Figure 1 -Overview of the FA-supplementation strategy for NP discovery. a) Cyanobacteria can elongate exogenous small-and medium-chain FAs and do not appear to carry out beta-oxidation; therefore, supplementation with deuterium-labeled hexanoic acid (as an example) will result in retention of the label in lipids incorporating longer-chain FAs, while in other bacteria (such as E. coli), exogenous FAs are not elongated and undergo beta-oxidative degradation.…”

Section: Introductionmentioning

confidence: 99%

Discovery of Cyanobacterial Natural Products Containing Fatty Acid Residues**

et al. 2021

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In recent years,extensive sequencing and annotation of bacterial genomes has revealed an unexpectedly large number of secondary metabolite biosynthetic gene clusters whose products are yet to be discovered. Fore xample, cyanobacterial genomes contain avariety of gene clusters that likely incorporate fatty acid derived moieties,b ut for most cases we lack the knowledge and tools to effectively predict or detect the encoded natural products.H ere,w ee xploit the apparent absence of af unctional b-oxidation pathway in cyanobacteria to achieve efficient stable-isotope-labeling of their fatty acid derived lipidome.W es how that supplementation of cyanobacterial cultures with deuterated fatty acids can be used to easily detect natural product signatures in individual strains.T he utility of this strategy is demonstrated in two cultured cyanobacteria by uncovering analogues of the multidrug-resistance reverting hapalosin, and novel, cytotoxic, lactylate-nocuolin Ah ybrids-the nocuolactylates.

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BrtB is an O-alkylating enzyme that generates fatty acid-bartoloside esters

Cited by 16 publications

References 32 publications

Biosynthesis of Chlorinated Lactylates in Sphaerospermopsis sp. LEGE 00249

Biosynthesis of Chlorinated Lactylates in Sphaerospermopsis sp. LEGE 00249

Distribution and diversity of dimetal-carboxylate halogenases in cyanobacteria

Discovery of Cyanobacterial Natural Products Containing Fatty Acid Residues**

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