Heterologous expression of cryptomaldamide in a cyanobacterial host

Taton, Arnaud; Ecker, Andrew K.; Diaz, Brienna; Moss, Nathan A.; Anderson, Brooke; Reher, Raphael; Leão, Tiago; Simkovsky, Ryan; Dorrestein, Pieter C.; Gerwick, Lena; Gerwick, William H.; Golden, James W.

doi:10.1101/2020.08.26.267179

Cited by 6 publications

(10 citation statements)

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“…It has been proposed that the selection of strains pre-adapted for growth in industrial photobioreactors could enable more productive, and therefore more economical, cultivation outcomes. Non-model filamentous cyanobacteria, including Arthrospira platensis (Spirulina), are becoming increasingly important for industrial bioproduction and biological engineering applications (Taton et al, 2020). This is a result of their capacity to form dense biofilms and, in some cases, their natural resilience to contamination due to unimpeded growth under high salinity and alkalinity conditions (Nies et al, 2017;Ataeian et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

The survivor strain: isolation and characterization of Phormidium yuhuli AB48, a filamentous phototactic cyanobacterium with biotechnological potential

Koch

Noonan

Qiu

et al. 2022

Front. Bioeng. Biotechnol.

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Despite their recognized potential, current applications of cyanobacteria as microbial cell factories remain in early stages of development. This is partly due to the fact that engineered strains are often difficult to grow at scale. This technical challenge contrasts with the dense and highly productive cyanobacteria populations thriving in many natural environments. It has been proposed that the selection of strains pre-adapted for growth in industrial photobioreactors could enable more productive cultivation outcomes. Here, we described the initial morphological, physiological, and genomic characterization of Phormidium yuhuli AB48 isolated from an industrial photobioreactor environment. P. yuhuli AB48 is a filamentous phototactic cyanobacterium with a growth rate comparable to Synechocystis sp. PCC 6803. The isolate forms dense biofilms under high salinity and alkaline conditions and manifests a similar nutrient profile to Arthrospira platensis (Spirulina). We sequenced, assembled, and analyzed the P. yuhuli AB48 genome, the first closed circular isolate reference genome for a member of the Phormidium genus. We then used cultivation experiments in combination with proteomics and metabolomics to investigate growth characteristics and phenotypes related to industrial scale cultivation, including nitrogen and carbon utilization, salinity, and pH acclimation, as well as antibiotic resistance. These analyses provide insight into the biological mechanisms behind the desirable growth properties manifested by P. yuhuli AB48 and position it as a promising microbial cell factory for industrial-scale bioproduction[221, 1631].

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Section: Introductionmentioning

confidence: 99%

The survivor strain: isolation and characterization of Phormidium yuhuli AB48, a filamentous phototactic cyanobacterium with biotechnological potential

Koch

Noonan

Qiu

et al. 2022

Front. Bioeng. Biotechnol.

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“…The M. bouillonii PNG columbamide BGC was amplified by PCR from genomic DNA as six overlapping fragments of 3 to 6 kb and cloned in one step into pAM5558 (Table S1), which carries sequences flanking the columbamide BGC, by TAR in the yeast Saccharomyces cerevisiae VL6-48N (Figure 2). 36,38 Positive yeast clones that contained the columbamide BGC were identified by PCR, and these plasmid constructs were transferred into E. coli. Plasmid preparation and analysis by NcoI restriction digests of a dozen clones identified 10 positive clones (Figure S2).…”

Section: Heterologous Expression Of the Columbamide Bgc Inmentioning

confidence: 99%

“…As we previously reported for the heterologous expression of the cryptomaldamide pathway from Moorena producens JHB, 36 S. elongatus failed to produce cryptomaldamide, but production in Anabaena was successful. Therefore, the columbamide BGC was next transferred into Anabaena.…”

Section: Heterologous Expression Of the Columbamide Bgc Inmentioning

confidence: 99%

“…32−34 To circumvent these limitations, the lyngbyatoxin A and cryptomaldamide BGCs were expressed in the well-studied cyanobacterium Anabaena (Nostoc) PCC 7120 (hereafter, Anabaena). 35,36 The lyngbyatoxin A pathway was further engineered in Anabaena to produce pendolmycin and teleocidin B-4. 37 Finally, the pathway encoded by a 42 kb BGC responsible for production of hapalindole alkaloids by the cyanobacterium Fischerella ambigua UTEX 1903 was engineered into Synechococcus elongatus UTEX 2973.…”

Section: ■ Introductionmentioning

confidence: 99%

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Heterologous Expression in Anabaena of the Columbamide Pathway from the Cyanobacterium Moorena bouillonii and Production of New Analogs

et al. 2022

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Columbamides are chlorinated acyl amide natural products, several of which exhibit cannabinomimetic activity. These compounds were originally discovered from a culture of the filamentous marine cyanobacterium Moorena bouillonii PNG5-198 collected from the coastal waters of Papua New Guinea. The columbamide biosynthetic gene cluster (BGC) had been identified using bioinformatics, but not confirmed by experimental evidence. Here, we report the heterologous expression in Anabaena (Nostoc) PCC 7120 of the 28.5 kb BGC that encodes for columbamide biosynthesis. The production of columbamides in Anabaena is investigated under several different culture conditions, and several new columbamide analogs are identified by liquid chromatography−tandem mass spectrometry (LC−MS/MS) and nuclear magnetic resonance (NMR). In addition to previously characterized columbamides A, B, and C, new columbamides I−M are produced in these experiments, and the structure of the most abundant monochlorinated analog, columbamide K (11), is fully characterized. The other new columbamide analogs are produced in only small quantities, and structures are proposed based on high-resolution-MS, MS/MS, and 1 H NMR data. Overexpression of the pathway's predicted halogenases resulted in increased productions of di-and trichlorinated compounds. The most significant change in production of columbamides in Anabaena is correlated with the concentration of NaCl in the medium.

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“…In another study, the neutral site 2 from S. elongatus PCC 7942 was introduced in Anabaena sp. PCC 7120 genome for direct tool transferability without requiring additional cloning steps (Taton et al, 2020), allowing introduction of a cryptomaldamide gene cluster from a marine cyanobacterium in both species. Interestingly, cryptomaldamide was only detected in Anabaena, hypothesized to be more suitable for production of complex secondary metabolites.…”

Section: The Genetic Toolkit For Freshwater Cyanobacteria Is More Ext...mentioning

confidence: 99%

Synthetic biology in marine cyanobacteria: Advances and challenges

Bourgade

Stensjö

2022

Front. Microbiol.

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The current economic and environmental context requests an accelerating development of sustainable alternatives for the production of various target compounds. Biological processes offer viable solutions and have gained renewed interest in the recent years. For example, photosynthetic chassis organisms are particularly promising for bioprocesses, as they do not require biomass-derived carbon sources and contribute to atmospheric CO2 fixation, therefore supporting climate change mitigation. Marine cyanobacteria are of particular interest for biotechnology applications, thanks to their rich diversity, their robustness to environmental changes, and their metabolic capabilities with potential for therapeutics and chemicals production without requiring freshwater. The additional cyanobacterial properties, such as efficient photosynthesis, are also highly beneficial for biotechnological processes. Due to their capabilities, research efforts have developed several genetic tools for direct metabolic engineering applications. While progress toward a robust genetic toolkit is continuously achieved, further work is still needed to routinely modify these species and unlock their full potential for industrial applications. In contrast to the understudied marine cyanobacteria, genetic engineering and synthetic biology in freshwater cyanobacteria are currently more advanced with a variety of tools already optimized. This mini-review will explore the opportunities provided by marine cyanobacteria for a greener future. A short discussion will cover the advances and challenges regarding genetic engineering and synthetic biology in marine cyanobacteria, followed by a parallel with freshwater cyanobacteria and their current genetic availability to guide the prospect for marine species.

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Heterologous expression of cryptomaldamide in a cyanobacterial host

Cited by 6 publications

References 80 publications

The survivor strain: isolation and characterization of Phormidium yuhuli AB48, a filamentous phototactic cyanobacterium with biotechnological potential

The survivor strain: isolation and characterization of Phormidium yuhuli AB48, a filamentous phototactic cyanobacterium with biotechnological potential

Heterologous Expression in Anabaena of the Columbamide Pathway from the Cyanobacterium Moorena bouillonii and Production of New Analogs

Synthetic biology in marine cyanobacteria: Advances and challenges

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