Production of ω3 fatty acids in marine cyanobacterium Synechococcus sp. strain NKBG 15041c via genetic engineering

Yoshino, Tomoko; Kakunaka, Natsumi; Yue, Liang; Ito, Yasuhito; Maeda, Yoshiaki; Nomaguchi, Tatsuhiro; Matsunaga, Tadashi

doi:10.1007/s00253-017-8407-1

Cited by 19 publications

(17 citation statements)

References 46 publications

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“…NKBG 15041c [62] and Ss7002 [43]. The first two produced high levels of ALA (23 and 50% of total FA, respectively) when engineered to overexpress, respectively, Δ6 + Δ15 and Δ6 FA desaturase genes [63, 64]. On the other hand, Ss7002 produced the highest ALA levels (19% of total FA) in a cyanobacterial wt strain when grown at high light intensity either at 22 °C or at 38 °C and shifted to 22 °C for 12 h [43].…”

Section: Discussionmentioning

confidence: 99%

Engineering the fatty acid synthesis pathway in Synechococcus elongatus PCC 7942 improves omega-3 fatty acid production

Santos-Merino

Garcillán-Barcia

Cruz

2018

Biotechnol Biofuels

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BackgroundThe microbial production of fatty acids has received great attention in the last few years as feedstock for the production of renewable energy. The main advantage of using cyanobacteria over other organisms is their ability to capture energy from sunlight and to transform CO2 into products of interest by photosynthesis, such as fatty acids. Fatty acid synthesis is a ubiquitous and well-characterized pathway in most bacteria. However, the activity of the enzymes involved in this pathway in cyanobacteria remains poorly explored.ResultsTo characterize the function of some enzymes involved in the saturated fatty acid synthesis in cyanobacteria, we genetically engineered Synechococcus elongatus PCC 7942 by overexpressing or deleting genes encoding enzymes of the fatty acid synthase system and tested the lipid profile of the mutants. These modifications were in turn used to improve alpha-linolenic acid production in this cyanobacterium. The mutant resulting from fabF overexpression and fadD deletion, combined with the overexpression of desA and desB desaturase genes from Synechococcus sp. PCC 7002, produced the highest levels of this omega-3 fatty acid.ConclusionsThe fatty acid composition of S. elongatus PCC 7942 can be significantly modified by genetically engineering the expression of genes coding for the enzymes involved in the first reactions of fatty acid synthesis pathway. Variations in fatty acid composition of S. elongatus PCC 7942 mutants did not follow the pattern observed in Escherichia coli derivatives. Some of these modifications can be used to improve omega-3 fatty acid production. This work provides new insights into the saturated fatty acid synthesis pathway and new strategies that might be used to manipulate the fatty acid content of cyanobacteria.Electronic supplementary materialThe online version of this article (10.1186/s13068-018-1243-4) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Engineering the fatty acid synthesis pathway in Synechococcus elongatus PCC 7942 improves omega-3 fatty acid production

Santos-Merino

Garcillán-Barcia

Cruz

2018

Biotechnol Biofuels

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“…While Vipp1 is reported to promote thylakoid membrane biogenesis and maintenance in plants and cyanobacteria associated with enhanced photosynthetic machinery (48,49), the effect of this gene on PUFA and LC-PUFA content has not been previously reported. It is well established that n-3 PUFA contents of cyanobacteria are significantly affected by temperature and light intensity (47,(50)(51)(52). The SDA and ETA contents may therefore be even further enhanced in these cyanobacteria by growth at lower temperatures and higher light intensities and further engineering of the cyanobacteria to enhance their capacity to generate more precursor substrates (LA, GLA or ALA) necessary for SDA and ETA production.…”

Section: Discussionmentioning

confidence: 99%

“…2 health-promoting EPA in humans, fish and livestock, a great deal of effort has gone into finding natural systems and designing new engineered pathways that produce high quantities of SDA and high ratios of n-3 to n-6 PUFAs (55). SDA is seldom naturally found in cyanobacteria (56) unless at least one acyl-lipid desaturase is provided on a multicopy vector (47), and even then, the maximal content of 26.6% SDA reached in the current study is as we have noted at least twofold higher than previous engineered strains (Table 3) (46,47). SDA-producing transgenic soybean oil contains 20-26% of the total fatty acids as SDA with n-3 to n-6 PUFA ratios of ~ 1 or lower (19,21,55,57), and seed oil from Echium plantagineum naturally contains ~13% of the total fatty acids as SDA (15,58,59).…”

Section: Discussionmentioning

confidence: 99%

Acyl-lipid desaturases and Vipp1 cooperate in cyanobacteria to produce novel omega-3 PUFA-containing glycolipids

Poole

Parsonage

Sergeant

et al. 2020

Preprint

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BackgroundDietary omega-3 (n-3), long chain (LC-, ≥ 20 carbons), polyunsaturated fatty acids (PUFAs) derived largely from marine animal sources protect against inflammatory processes and enhance brain development and function. With the depletion of natural stocks of marine animal sources and an increasing demand for n-3 LC-PUFAs, alternative, sustainable supplies are urgently needed. As a result, n-3 18 carbon and LC-PUFAs are being generated from plant or algal sources, either by engineering new biosynthetic pathways or by augmenting existing systems.ResultsWe utilized an engineered plasmid encoding two cyanobacterial acyl-lipid desaturases (DesB and DesD, encoding Δ15 and Δ6 desaturases, respectively) and “vesicle-inducing protein in plastids” (Vipp1) to induce production of stearidonic acid (SDA,18:4 n-3) at high levels in three strains of cyanobacteria (10, 17 and 27% of total lipids in Anabaena sp. PCC7120, Synechococcus sp. PCC7002, and Leptolyngbya sp. strain BL0902, respectively). Lipidomic analysis revealed that in addition to SDA, the rare anti-inflammatory n-3 LC-PUFA eicosatetraenoic acid (ETA, 20:4 n-3) was synthesized in these engineered strains, and ∼99% of SDA and ETA was complexed to bioavailable monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) species. Importantly, novel molecular species containing alpha-linolenic acid (ALA), SDA and/or ETA in both acyl positions of MGDG and DGDG were observed in the engineered Leptolyngbya and Synechococcus strains, suggesting that these could provide a rich source of anti-inflammatory molecules.ConclusionsOverall, this technology utilizes solar energy, consumes carbon dioxide, and produces large amounts of nutritionally-important n-3 PUFAs and LC-PUFAs. Importantly, it can generate previously-undescribed, highly bioavailable, anti-inflammatory galactosyl lipids. This technology could therefore be transformative in protecting ocean fisheries and augmenting the nutritional quality of human and animal food products.Broader ContextDietary omega-3 (n-3), long chain polyunsaturated fatty acids (LC-PUFAs) typically found in marine products such as fish and krill oil are beneficial to human health. In addition to human consumption, most of the global supply of n-3 LC-PUFAs is used as dietary components for aquaculture. Marked increases in usage have created an intense demand for more sustainable, stable and bioavailable forms of n-3 PUFAs and LC-PUFAs. We utilized an engineered plasmid to dramatically enhance the production of 18-carbon and n-3 LC-PUFAs in three strains of autotrophic cyanobacteria. While the sustainable generation of highly valued and bioavailable nutritional lipid products is the primary goal, additional benefits include the generation of oxygen as a co-product with the consumption of only carbon dioxide as the carbon source and solar radiation as the energy source. This technology could be transformative in protecting ocean fisheries and augmenting the nutritional quality of human and animal food products. Additionally, these engineered cyanobacteria can generate previously undescribed, highly bioavailable, anti-inflammatory galactosyl lipids.

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“…Most of the cyanobacteria have a fatty acid profile dominated by C14, C16 and C18 saturated and monounsaturated fatty acids. However, quite often they contain significant quantities of some essential polyunsaturated fatty acids such as C18 linoleic (18:2ω6) and α-linolenic (18:3ω3) acids whose production in some cases can reach up to 20% of the cellular dry weight [18]. The FAME profiles of Synechococcus sp.…”

Section: Discussionmentioning

confidence: 99%

“…In cyanobacteria, fatty acids are doubly important, as membrane lipids and as constituents of glycolipids, which form the thylakoid membranes where photosynthesis takes place. Cyanobacteria often contain significant quantities of some essential polyunsaturated fatty acids (PUFAs) such as C18 linoleic (18:2ω6) and α-linolenic (18:3ω3) acids whose production can reach up to 20% of the cellular dry weight via genetic engineering [18]. The species rich in saturated fatty acids (SFAs) are good candidates for biofuel production whereas species rich in PUFAs are considered more appropriate for production of nutrition supplements, animal feed, etc.…”

Section: Introductionmentioning

confidence: 99%

Growth Aspects and Biochemical Composition of Synechococcus sp. MK568070 Cultured in Oil Refinery Wastewater

Blažina

Haberle

Hrustić

et al. 2019

JMSE

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The aim of the study was to explore the possibility of bioremediation of oil refinery wastewaters by the cyanobacterium Synechococcus sp. MK568070, isolated from the Adriatic Sea. The potential of biomass and lipid production was explored upon cultivation on oil refinery wastewater with excess CO2 after the removal of nutrients. The strain grew well in a wide range of salinities and ammonium concentrations, and was further tested on the wastewater from local oil refinery plant of various N-composition. Growth experiment under optimized conditions was used to analyze the lipid, carbohydrate and protein dynamics. The biomass yield was highly dependent on nutrient source and concentration, salinity and CO2 addition. Highest biomass yield was 767 mg/L of dry weight. Towards the end of the experiment the decline in carbohydrate to 18.9% is visible, whereas at the same point lipids, in particular saturated fatty acid methyl esters (FAME), started to accumulate within the cells. The content of lipids at the end of the experiment was 21.4%, with the unsaturation index 0.45 providing good biofuel feedstock characteristics. Fourier Transform Infrared (FTIR) spectroscopy analysis demonstrated a high degree of lipid accumulation in respect to proteins, along with the structural changes and biomass accumulation. In addition, the N-removal from the wastewater was >99% efficient. The potential of lipid accumulation, due to the functional photosynthesis even at the minimal cell quota of nutrients, is critical for the usage of excess industrial CO2 and its industrial transformation to biodiesel. These findings enable further considerations of Synechococcus sp. (MK568070) for the industrial scale biomass production and wastewater remediation.

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Production of ω3 fatty acids in marine cyanobacterium Synechococcus sp. strain NKBG 15041c via genetic engineering

Cited by 19 publications

References 46 publications

Engineering the fatty acid synthesis pathway in Synechococcus elongatus PCC 7942 improves omega-3 fatty acid production

Engineering the fatty acid synthesis pathway in Synechococcus elongatus PCC 7942 improves omega-3 fatty acid production

Acyl-lipid desaturases and Vipp1 cooperate in cyanobacteria to produce novel omega-3 PUFA-containing glycolipids

Growth Aspects and Biochemical Composition of Synechococcus sp. MK568070 Cultured in Oil Refinery Wastewater

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