Expanding the Natural Products Heterologous Expression Repertoire in the Model Cyanobacterium Anabaena sp. Strain PCC 7120: Production of Pendolmycin and Teleocidin B-4

Videau, Patrick; Wells, Kaitlyn N; Singh, Arun; Eiting, Jessie; Proteau, Philip; Philmus, Benjamin

doi:10.26434/chemrxiv.11316098.v1

Cited by 4 publications

(4 citation statements)

References 5 publications

(5 reference statements)

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“…32 Recently, the lyngbyatoxin pathway was further engineered in Anabaena to produce pendolmycin and teleocidin B-4. 33 The development of approaches for the heterologous expression of NP pathways is important to facilitate the characterization and screening of cyanobacterial NPs for pharmaceutical applications. Strains of the genus Moorena, previously named Moorea 34 , have been found to harbor over 40 different biosynthetic gene clusters and close to 200 novel NPs have been chemically identified from Moorena spp.…”

Section: Introductionmentioning

confidence: 99%

Heterologous Expression of Cryptomaldamide in a Cyanobacterial Host

et al. 2020

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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.

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Section: Introductionmentioning

confidence: 99%

Heterologous Expression of Cryptomaldamide in a Cyanobacterial Host

et al. 2020

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“…This has impeded not only new compound discovery, but also exploration of its genomic characteristics, including those that are responsible for natural product biosynthesis. It is possible that developments with the heterologous expression of cyanobacterial natural product pathways will enable a more extensive exploration of the rich secondary metabolome of this genus in the future [ 91 , 92 ].…”

Section: Discussionmentioning

confidence: 99%

The Chemistry, Biochemistry and Pharmacology of Marine Natural Products from Leptolyngbya, a Chemically Endowed Genus of Cyanobacteria

Naman

Alexander

et al. 2020

Marine Drugs

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Leptolyngbya, a well-known genus of cyanobacteria, is found in various ecological habitats including marine, fresh water, swamps, and rice fields. Species of this genus are associated with many ecological phenomena such as nitrogen fixation, primary productivity through photosynthesis and algal blooms. As a result, there have been a number of investigations of the ecology, natural product chemistry, and biological characteristics of members of this genus. In general, the secondary metabolites of cyanobacteria are considered to be rich sources for drug discovery and development. In this review, the secondary metabolites reported in marine Leptolyngbya with their associated biological activities or interesting biosynthetic pathways are reviewed, and new insights and perspectives on their metabolic capacities are gained.

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“…In some marine bacteria, PKS synthesize very long chain fatty acids, such as docosahexaenoic acids (Okuyama et al, 2007). Recently, NRPSs have successfully been expressed in Anabaena for bioproduction (Videau et al, 2016(Videau et al, , 2019. The replacement of genes involved in the synthesis of Hgl with desired PKS/NRPS will be of great interest.…”

Section: Anabaena Is a Useful Platform For Bioproductionmentioning

confidence: 99%

hetN and patS Mutations Enhance Accumulation of Fatty Alcohols in the hglT Mutants of Anabaena sp. PCC 7120

2020

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The heterocysts present in filamentous cyanobacteria such as Anabaena sp. PCC 7120 are known to be regulated by HetN and PatS, the repressors of heterocyst differentiation; therefore, the inactivation of these proteins will result in the formation of multiple heterocysts. To enhance the accumulation of fatty alcohols synthesized in the heterocyst, we introduced mutations of these repressors to increase heterocyst frequency. First, we isolated double mutants of hetN and patS and confirmed that the null mutation of these genes promoted higher frequencies of heterocyst formation and higher accumulation of heterocyst-specific glycolipids (Hgls) compared with its wild type. Next, we combined hetN and patS mutations with an hglT (encoding glycosyltransferase, an enzyme involved in Hgl synthesis) mutation to increase the accumulation of fatty alcohols since knockout mutation of hglT results in accumulation of very long chain fatty alcohol, the precursor of Hgl. We also observed retarded growth, lower chlorophyll content and up to a five-fold decrease in photosynthetic activity of the hetN / patS / hglT triple mutants. In contrast, the triple mutants showed three times higher heterocyst formation frequencies than the hglT single mutant and wild type. The production rate of fatty alcohol in the triple mutants attained a value 1.41 nmol/mL OD 730 , whereas accumulation of Hgls in the wild type was 0.90 nmol/mL OD 730 . Aeration of culture improved the accumulation of fatty alcohols in hetN / patS / hglT mutant cells up to 2.97 nmol/mL OD 730 compared with cells cultured by rotation. Our study outlines an alternative strategy for fatty alcohol production supported by photosynthesis and nitrogen fixation.

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Expanding the Natural Products Heterologous Expression Repertoire in the Model Cyanobacterium Anabaena sp. Strain PCC 7120: Production of Pendolmycin and Teleocidin B-4

Cited by 4 publications

References 5 publications

Heterologous Expression of Cryptomaldamide in a Cyanobacterial Host

Heterologous Expression of Cryptomaldamide in a Cyanobacterial Host

The Chemistry, Biochemistry and Pharmacology of Marine Natural Products from Leptolyngbya, a Chemically Endowed Genus of Cyanobacteria

hetN and patS Mutations Enhance Accumulation of Fatty Alcohols in the hglT Mutants of Anabaena sp. PCC 7120

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