Unlocking the Spatial Control of Secondary Metabolism Uncovers Hidden Natural Product Diversity in <i>Nostoc punctiforme</i>

Dehm, Daniel; Krumbholz, Julia; Baunach, Martin; Wiebach, Vincent; Hinrichs, K.; Guljamow, Arthur; Tabuchi, Takeshi; Jenke‐Kodama, Holger; Süssmuth, Roderich D.; Dittmann, Elke

doi:10.1021/acschembio.9b00240

Cited by 35 publications

(72 citation statements)

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“…High density cultivation (HDC) is a recently developed cultivation system that tackles these problems by implementing a membrane-mediated CO 2 supply technology, combined with optimised nutrient supply and high light intensities 27 . This system facilitates rapid, sustainable biomass accumulation and was demonstrated to further raise the potential for the recovery of natural products from cyanobacteria [28][29][30] .…”

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confidence: 99%

High density cultivation for efficient sesquiterpenoid biosynthesis in Synechocystis sp. PCC 6803

Dienst

Wichmann

Mantovani

et al. 2020

Sci Rep

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cyanobacteria and microalgae are attractive photoautotrophic host systems for climate-friendly production of fuels and other value-added biochemicals. However, for economic applications further development and implementation of efficient and sustainable cultivation strategies are essential. Here, we present a comparative study on cyanobacterial sesquiterpenoid biosynthesis in Synechocystis sp. PCC 6803 using a commercial lab-scale High Density Cultivation (HDC) platform in the presence of dodecane as in-situ extractant. operating in a two-step semi-batch mode over a period of eight days, volumetric yields of (E)-α-bisabolene were more than two orders of magnitude higher than previously reported for cyanobacteria, with final titers of 179.4 ± 20.7 mg * L −1. Likewise, yields of the sesquiterpene alcohols (−)-patchoulol and (−)-α-bisabolol were many times higher than under reference conditions, with final titers of 17.3 ± 1.85 mg * L −1 and 96.3 ± 2.2 mg * L −1 , respectively. While specific productivity was compromised particularly for (E)-α-bisabolene in the HDc system during phases of high biomass accumulation rates, volumetric productivity enhancements during linear growth at high densities were more pronounced for (E)-α-bisabolene than for the hydroxylated terpenoids. together, this study provides additional insights into cell density-related process characteristics, introducing HDC as highly efficient strategy for phototrophic terpenoid production in cyanobacteria. Cyanobacteria are oxygenic phototrophs with a versatile metabolism and therefore constitute an economically appealing platform for the sustainable production of a diversity of industrially relevant chemicals. In particular, the natural occurrence of the methylerythritol-4-phosphate (MEP) pathway 1 allows the engineered biosynthesis of a diversity of high-value terpenoid compounds such as pharmaceuticals, cosmetics, and biofuels (reviewed in 2,3). While numerous pioneering proof-of-concept studies already demonstrated the heterologous biosynthesis of isoprene (C 5), mono-(C 10), sesqui-(C 15), di-(C 20) and tri-(C 30) terpenes in cyanobacterial host strains 4-9 , the reported titers of these compounds are typically two to three orders of magnitudes lower than those of e.g. alcoholic fuels (cf. overviews in 3,10). The major limitation of these processes arise from the inherent carbon partitioning characteristics, which favour primary metabolism and biomass accumulation, resulting in weak carbon fluxes through the native MEP pathway 11. In accordance, a strategy to bypass these inherent flux limitations by heterologous expression of the complete bacterial mevalonate (MVA) pathway in Synechocystis sp. PCC 6803 (from here: Synechocystis) resulted in 2.5-fold higher isoprene yields 12. Likewise, numerous rational design studies targeting selected bottleneck steps within the MEP pathway as well as selected steps of upstream carbon fluxes lent support to this concept 8,9,13-19. Further strategies sought to challenge the problem of inherently weak terpene s...

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confidence: 99%

High density cultivation for efficient sesquiterpenoid biosynthesis in Synechocystis sp. PCC 6803

Dienst

Wichmann

Mantovani

et al. 2020

Sci Rep

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“…GSV224 [93], Nostoc sp. UK2almI isolated from lichen [94], and Nostoc punctiforme PCC73102 [95]. The general structure of the three Nps (A1–A3) from GSV224 is BA-IleA1/ValA2)[Ser-MePro-LeuAc-Leu-Gly-Asn-Tyr-Pro], where MePro and leucylacetate (LeuAc) are the two non-proteinogenic amino acids and BA is a butyryl group.…”

Section: Other Peptides Exclusively Produced By Nostocmentioning

confidence: 99%

“…As many other cyanobacteria, Nostoc produces bioactive nonribosomal peptides such as microcystins, cyanopeptolins, microginins, and anabaenopeptins. There are also reports on the presence of the rybosomaly synthesized microviridins in Nostoc minutum NIES-26 [100] and N. punctiform e PCC73102 [95] (Table S1). The structure, the activity, and the biosynthetic pathways of the peptides have been described in many scientific papers (e.g., [28,32,101,102]).…”

Section: Peptides Produced By Nostoc and Other Cyanobacteriamentioning

confidence: 99%

Bioactive Peptides Produced by Cyanobacteria of the Genus Nostoc: A Review

2019

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Cyanobacteria of the genus Nostoc are widespread in all kinds of habitats. They occur in a free-living state or in association with other organisms. Members of this genus belong to prolific producers of bioactive metabolites, some of which have been recognized as potential therapeutic agents. Of these, peptides and peptide-like structures show the most promising properties and are of a particular interest for both research laboratories and pharmaceutical companies. Nostoc is a sole source of some lead compounds such as cytotoxic cryptophycins, antiviral cyanovirin-N, or the antitoxic nostocyclopeptides. Nostoc also produces the same bioactive peptides as other cyanobacterial genera, but they frequently have some unique modifications in the structure. This includes hepatotoxic microcystins and potent proteases inhibitors such as cyanopeptolins, anabaenopeptins, and microginins. In this review, we described the most studied peptides produced by Nostoc, focusing especially on the structure, the activity, and a potential application of the compounds.

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“…16 Transcriptomics and metabolomics along with genome sequence bioinformatics have also revealed that a large fraction of these BGCs are not expressed in standard laboratory cultures, leaving a substantial fraction of diverse cyanobacterial natural product compounds unexplored. 17 Recent studies have shown that altered growth conditions can result in upregulation of some cryptic BGCs; 17,18 however, this approach still leaves many BGCs unexpressed in laboratory growth conditions.…”

Section: Introductionmentioning

confidence: 99%

Heterologous expression of cryptomaldamide in a cyanobacterial host

Taton

Ecker

Diaz

et al. 2020

Preprint

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Filamentous marine cyanobacteria make a variety of bioactive molecules that are produced by polyketide synthases, non-ribosomal peptide synthetases, and hybrid pathways that are encoded by large biosynthetic gene clusters. These cyanobacterial natural products represent potential drugs leads; however, thorough pharmacological investigations have been impeded by the limited quantity of compound that is typically available from the native organisms. Additionally, investigations of the biosynthetic gene clusters and enzymatic pathways have been difficult due to the inability to conduct genetic manipulations in the native producers. Here we report a set of genetic tools for the heterologous expression of biosynthetic gene clusters in the cyanobacteria Synechococcus elongatus PCC 7942 and Anabaena (Nostoc) PCC 7120. To facilitate the transfer of gene clusters in both strains, we engineered a strain of Anabaena that contains S. elongatus homologous sequences for chromosomal recombination at a neutral site and devised a CRISPR-based strategy to efficiently obtain segregated double recombinant clones of Anabaena. These genetic tools were used to express the large 28.7 kb cryptomaldamide biosynthetic gene cluster from the marine cyanobacterium Moorena (Moorea) producens JHB in both model strains. S. elongatus did not produce cryptomaldamide, however high-titer production of cryptomaldamide was obtained in Anabaena. The methods developed in this study will facilitate the heterologous expression of biosynthetic gene clusters isolated from marine cyanobacteria and complex metagenomic samples.Abstract Figure

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Unlocking the Spatial Control of Secondary Metabolism Uncovers Hidden Natural Product Diversity in Nostoc punctiforme

Cited by 35 publications

References 40 publications

High density cultivation for efficient sesquiterpenoid biosynthesis in Synechocystis sp. PCC 6803

High density cultivation for efficient sesquiterpenoid biosynthesis in Synechocystis sp. PCC 6803

Bioactive Peptides Produced by Cyanobacteria of the Genus Nostoc: A Review

Heterologous expression of cryptomaldamide in a cyanobacterial host

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