Cyanobacterial Sfp-type phosphopantetheinyl transferases functionalize carrier proteins of diverse biosynthetic pathways

Yang, Guang; Zhang, Yi; Lee, Nicholas K.; Cozad, Monica A.; Kearney, Sara E.; Luesch, Hendrik; Ding, Yousong

doi:10.1038/s41598-017-12244-3

Cited by 18 publications

(17 citation statements)

References 43 publications

(51 reference statements)

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“…5, soluble protein extracts from the recombinant cyanobacterial strains pursued in this study allowed the conversion of E. coli ACP to its fluorescently labeled derivative. These results indicate that S. elongatus wild-type strain AMC2302 does not exhibit promiscuous native PPTase activity, as was previously reported 52 , and that the heterologous Sfp is expressed in its active form in all the S. elongatus recombinant strains analyzed. Therefore, considering that we had previously shown that the Mas synthase is a suitable substrate for Sfp 8,55 , we were confident that the Mas enzyme would also be adequately phosphopantetheinylated in the recombinant strains here developed.…”

Section: Resultssupporting

confidence: 87%

“…The S. elongatus genome does not encode PKS or non-ribosomal peptide synthase (NRPS) clusters 50,51 . Although a Sfp-type PPTase was identified in its genome 52 , further characterization of this PPTase showed that it exhibits very low or no activity towards ACPs of different PKSs 52 . Therefore, as part of module II, we constructed a device in which the promiscuous Sfp-PPTase from Bacillus subtilis 49 was expressed from P conII and placed under the translational control of riboswitch F (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Specialized natural product pathways, in particular PKS and NRPS, invariably rely on the post-translational modification of the synthases to produce active enzymes. Because S. elongatus lacks a PPTase with activity towards the carrier proteins of different PKSs or NRPSs 52 , a promiscuous PPTase enzyme 49 was included in our platform strain, and in vitro assays confirmed that the enzyme was active (Fig. 3 and Supplementary Fig.…”

Section: Discussionmentioning

confidence: 99%

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Development of a cyanobacterial heterologous polyketide production platform

Roulet

Taton

Golden

et al. 2018

Metabolic Engineering

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The development of new heterologous hosts for polyketides production represents an excellent opportunity to expand the genomic, physiological, and biochemical backgrounds that better fit the sustainable production of these valuable molecules. Cyanobacteria are particularly attractive for the production of natural compounds because they have minimal nutritional demands and several strains have well established genetic tools. Using the model strain Synechococcus elongatus, a generic platform was developed for the heterologous production of polyketide synthase (PKS)-derived compounds. The versatility of this system is based on interchangeable modules harboring promiscuous enzymes for PKS activation and the production of PKS extender units, as well as inducible circuits for a regulated expression of the PKS biosynthetic gene cluster. To assess the capability of this platform, we expressed the mycobacterial PKS-based mycocerosic biosynthetic pathway to produce multimethyl-branched esters (MBE). This work is a foundational step forward for the production of high value polyketides in a photosynthetic microorganism.

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Section: Resultssupporting

confidence: 87%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Development of a cyanobacterial heterologous polyketide production platform

Roulet

Taton

Golden

et al. 2018

Metabolic Engineering

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“…For example, the gene of a promiscuous PPTase from the myxobacterium Stigmatella aurantiaca (MtaA) was integrated into the chromosome of E. coli GB2005 to create E. coli GB05-MtaA, which successfully expressed the microcystin BGC to produce multiple analogs (Liu et al., 2017 , 2019 ). To gain useful insights into the catalytic performance of cyanobacterial PPTases, we recently characterized the substrate scopes of 6 enzymes with 11 thiolation domains of known and silent BGCs from cyanobacterial and Streptomyces strains (Yang et al., 2017 ). Biochemical and genetic studies uncovered that the PPTase from Anabaena sp.…”

Section: Metabolic Engineering and Synthetic Biology Toolkits For Heterologous Production Of Cyanobacterial Compoundsmentioning

confidence: 99%

Heterologous production of cyanobacterial compounds

Dhakal

Chen

Luesch

et al. 2021

Journal of Industrial Microbiology and Biotechnology

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Cyanobacteria produce a plethora of compounds with unique chemical structures and diverse biological activities. Importantly, the increasing availability of cyanobacterial genome sequences and the rapid development of bioinformatics tools have unraveled the tremendous potential of cyanobacteria in producing new natural products. However, the discovery of these compounds based on cyanobacterial genomes has progressed slowly as the majority of their corresponding biosynthetic gene clusters (BGCs) are silent. In addition, cyanobacterial strains are often slow-growing, difficult for genetic engineering, or cannot be cultivated yet, limiting the use of host genetic engineering approaches for discovery. On the other hand, genetically tractable hosts such as Escherichia coli, Actinobacteria, and yeast have been developed for the heterologous expression of cyanobacterial BGCs. More recently, there has been increased interests in developing model cyanobacterial strains as heterologous production platforms. Herein, we present recent advances in the heterologous production of cyanobacterial compounds in both cyanobacterial and non-cyanobacterial hosts. Emerging strategies for BGC assembly, host engineering, and optimization of BGC expression are included for fostering the broader applications of synthetic biology tools in the discovery of new cyanobacterial natural products.

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“…BGCs usually include genes for core biosynthesis and tailoring enzymes and regulatory and resistance genes [6,7]. Among the accessory enzymes, 4-phosphopantetheinyl transferases (PPTs) play a major role in the biosynthesis of several natural products through the conversion of inactive apo-proteins into their active holo-forms [8][9][10].…”

Section: Introductionmentioning

confidence: 99%

Mining of Cyanobacterial Genomes Indicates That Plasmids Are Involved in the Production of Natural Products

Popin

Alvarenga

Castelo-Branco

et al. 2020

Preprint

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Background Microbial natural products have unique chemical structures and diverse biological activities. Cyanobacteria commonly possess a wide range of biosynthetic gene clusters to produce natural products. Several studies have mapped the distribution of natural product biosynthetic gene clusters in cyanobacterial genomes. However, little attention has been paid to natural product biosynthesis in plasmids. Some genes encoding cyanobacterial natural product biosynthetic pathways are believed to be dispersed by plasmids through horizontal gene transfer. Thus, we examined complete cyanobacterial genomes to assess if plasmids are involved in the production and dissemination of natural products by cyanobacteria.Results The 185 analyzed genomes possessed 1 to 42 gene clusters and an average of 10. In total, 1816 biosynthetic gene clusters were found. Approximately 95% of these clusters were present in chromosomes. The remaining 5% were present in plasmids, from which homologs of the biosynthetic pathways for aeruginosin, anabaenopeptin, ambiguine, cryptophycin, hassallidin, geosmin, and microcystin were manually curated. The cryptophycin pathway was previously described as active while the other gene cluster include all genes for biosynthesis. Approximately 12% of the 424 analyzed cyanobacterial plasmids contained homologs of genes involved in conjugation. Large plasmids, previously named as “chromids”, were also observed to be widespread in cyanobacteria. Sixteen cryptic natural product biosynthetic gene clusters and geosmin biosynthetic gene clusters were located in those mobile plasmids.Conclusion Homologues of genes involved in the production of toxins, protease inhibitors, odorous compounds, antimicrobials, antitumorals, and other unidentified natural products are located in cyanobacterial plasmids. Some of these plasmids are predicted to be conjugative. The present study provides in silico evidence that plasmids are involved in the distribution of natural product biosynthetic pathways in cyanobacteria.

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Cyanobacterial Sfp-type phosphopantetheinyl transferases functionalize carrier proteins of diverse biosynthetic pathways

Cited by 18 publications

References 43 publications

Development of a cyanobacterial heterologous polyketide production platform

Development of a cyanobacterial heterologous polyketide production platform

Heterologous production of cyanobacterial compounds

Mining of Cyanobacterial Genomes Indicates That Plasmids Are Involved in the Production of Natural Products

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